Narcissus and Daffodil

Narcissus and Daffodil

Medicinal and Aromatic Plants – Industrial Profiles 

Individual volumes in this series provide both industry and academia with in-depth coverage of one 

major medicinal or aromatic plant of industrial importance. 

Edited by Dr Roland Hardman 

Volume 1 

Valerian, edited by Peter J. Houghton 

Volume 2 

Perilla, edited by He-ci Yu, Kenichi Kosuna and Megumi Haga 

Volume 3 

Poppy, edited by Jenö Bernáth 

Volume 4 

Cannabis, edited by David T. Brown 

Volume 5 

Neem, edited by H.S. Puri 

Volume 6 

Ergot, edited by Vladimír Kren and Ladislav Cvak 

Volume 7 

Caraway, edited by Éva Németh 

Volume 8 

Saffron, edited by Moshe Negbi 

Volume 9 

Tea Tree, edited by Ian Southwell and Robert Lowe 

Volume 10 

Basil, edited by Raimo Hiltunen and Yvonne Holm 

Volume 11 

Fenugreek, edited by Georgios Petropoulos 

Volume 12 

Gingko biloba, edited by Teris A. Van Beek 

Volume 13 

Black Pepper, edited by P.N. Ravindran 

Volume 14 

Sage, edited by Spiridon E. Kintzios 

Volume 15 

Ginseng, edited by W.E. Court 

Volume 16 

Mistletoe, edited by Arndt Büssing 

Volume 17 

Tea, edited by Yong-su Zhen 

Volume 18 

Artemisia, edited by Colin W. Wright 

Volume 19 

Stevia, edited by A. Douglas Kinghorn 

Volume 20 

Vetiveria, edited by Massimo Maffei 

Volume 21 

Narcissus and Daffodil, edited by Gordon R. Hanks

Narcissus and Daffodil 

The genus Narcissus 

Edited by 

Gordon R. Hanks 

Horticulture Research International, Kirton, UK 

London and New York

First published 2002 

by Taylor & Francis 

11 New Fetter Lane, London EC4P 4EE 

Simultaneously published in the USA and Canada 

by Taylor & Francis Inc, 

29 West 35th Street, New York, NY 10001 

Taylor & Francis is an imprint of the Taylor & Francis Group 

© 2002 Taylor & Francis 

Typeset in Baskerville BT by 

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Printed and bound in Great Britain by 

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All rights reserved. No part of this book may be reprinted or reproduced 

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now known or hereafter invented, including photocopying and recording, 

or in any information storage or retrieval system, without permission in 

writing from the publishers. 

Every effort has been made to ensure that the advice and information 

in this book is true and accurate at the time of going to press. However, 

neither the publisher nor the authors can accept any legal responsibility or 

liability for any errors or omissions that may be made. In the case of drug 

administration, any medical procedure or the use of technical equipment 

mentioned within this book, you are strongly advised to consult the 

manufacturer’s guidelines. 

British Library Cataloguing in Publication Data 

A catalogue record for this book is available from the British Library 

Library of Congress Cataloging in Publication Data 

A catalog record for this book has been requested 

ISBN 0–415–27344–7

Contents 

Preface to the series vii 

Preface ix 

Acknowledgements xi 

List of contributors xii 

List of figures/plates xv 

1 The biology of Narcissus 1 

GORDON R. HANKS 

2 The folklore of Narcissus 19 

ANTHONY C. DWECK 

3 Classification of the genus Narcissus 30 

BRIAN MATHEW 

4 Commercial production of Narcissus bulbs 53 

GORDON R. HANKS 

5 Economics of Narcissus bulb production 131 

JAMES B. BRIGGS 

6 Alkaloids of Narcissus 141 

JAUME BASTIDA AND FRANCESC VILADOMAT 

7 Production of galanthamine by Narcissus tissues in vitro 215 

CARLES CODINA 

8 Narcissus and other Amaryllidaceae as sources of galanthamine 242 

O.A. CHERKASOV AND O.N. TOLKACHEV 

9 Studies on galanthamine extraction from Narcissus and 

other Amaryllidaceae 256 

MIRKO KREH

vi Contents 

10 Galanthamine production from Narcissus: agronomic 

and related considerations 273 

RITA M. MORAES 

11 Extraction and quantitative analysis of Amaryllidaceae alkaloids 286 

N.P. DHAMMIKA NANAYAKKARA AND JAIRO K. BASTOS 

12 Synthesis of galanthamine and related compounds 304 

V.N. BULAVKA AND O.N. TOLKACHEV 

13 Compounds from the genus Narcissus: 

pharmacology, pharmacokinetics and toxicology 332 

DAVID BROWN 

14 Galanthamine: clinical trials in Alzheimer’s disease 355 

DAVID BROWN 

15 Screening of Amaryllidaceae for biological activities: 

acetylcholinesterase inhibitors in Narcissus 369 

KORNKANOK INGKANINAN, HUBERTUS IRTH AND ROB VERPOORTE 

16 Narcissus lectins 380 

ELS J.M. VAN DAMME AND WILLY J. PEUMANS 

17 Narcissus in perfumery 392 

CHRISTIAN REMY 

18 Harmful effects due to Narcissus and its constituents 399 

CELIA G. JULIAN AND PETER W. BOWERS 

19 Review of pharmaceutical patents from the genus Narcissus 408 

JAMES R. MURRAY 

Index 419

Preface to the series 

There is increasing interest in industry, academia and the health sciences in 

medicinal and aromatic plants. In passing from plant production to the eventual 

product used by the public, many sciences are involved. This series brings 

together information which is currently scattered through an ever increasing 

number of journals. Each volume gives an in-depth look at one plant genus, about 

which an area specialist has assembled information ranging from the production 

of the plant to market trends and quality control. 

Many industries are involved such as forestry, agriculture, chemical, food, flavour, 

beverage, pharmaceutical, cosmetic and fragrance. The plant raw materials 

are roots, rhizomes, bulbs, leaves, stems, barks, wood, flowers, fruits and seeds. 

These yield gums, resins, essential (volatile) oils, fixed oils, waxes, juices, extracts 

and spices for medicinal and aromatic purposes. All these commodities are traded 

worldwide. A dealer’s market report for an item may say ‘Drought in the country 

of origin has forced up prices’. 

Natural products do not mean safe products and account of this has to be taken 

by the above industries, which are subject to regulation. For example, a number 

of plants which are approved for use in medicine must not be used in cosmetic 

products. 

The assessment of safe to use starts with the harvested plant material which has 

to comply with an official monograph. This may require absence of, or prescribed 

limits of, radioactive material, heavy metals, aflatoxin, pesticide residue, as well as 

the required level of active principle. This analytical control is costly and tends to 

exclude small batches of plant material. Large scale contracted mechanised 

cultivation with designated seed or plantlets is now preferable. 

Today, plant selection is not only for the yield of active principle, but for the 

plant’s ability to overcome disease, climatic stress and the hazards caused by mankind. 

Such methods as in vitro fertilisation, meristem cultures, and somatic 

embryogenesis are used. The transfer of sections of DNA is giving rise to controversy 

in the case of some end-uses of the plant material. 

Some suppliers of plant raw material are now able to certify that they are 

supplying organically-farmed medicinal plants, herbs and spices. The European 

Union directive (CVO/EU No 2092/91) details the specifications for the obligatory 

quality controls to be carried out at all stages of production and processing of 

organic products. 

Fascinating plant folklore and ethnopharmacology leads to medicinal potential. 

Examples are the muscle relaxants based on the arrow poison, curare, from

viii Preface to the series 

species of Chondrodendron, and the antimalarials derived from species of Cinchona 

and Artemisia. The methods of detection of pharmacological activity have become 

increasingly reliable and specific, frequently involving enzymes in bioassays and 

avoiding the use of laboratory animals. By using bioassay linked fractionation of 

crude plant juices or extracts, compounds can be specifically targeted which, for 

example, inhibit blood platelet aggregation, or have antitumour, or antiviral, or 

any other required activity. With the assistance of robotic devices, all the members 

of a genus may be readily screened. However, the plant material must be fully 

authenticated by a specialist. 

The medicinal traditions of ancient civilisations such as those of China and India 

have a large armamentarium of plants in their pharmacopoeias which are used 

throughout South East Asia. A similar situation exists in Africa and South America. 

Thus, a very high percentage of the World’s population relies on medicinal and 

aromatic plants for their medicine. Western medicine is also responding. Already 

in Germany all medical practitioners have to pass an examination in phytotherapy 

before being allowed to practise. It is noticeable that throughout Europe and 

the USA, medical, pharmacy and health related schools are increasingly offering 

training in phytotherapy. 

Multinational pharmaceutical companies have become less enamoured of the 

single compound magic bullet cure. The high costs of such ventures and the endless 

competition from “me too” compounds from rival companies often discourage 

the attempt. Independent phytomedicine companies have been very strong in 

Germany. However, by the end of 1995, eleven (almost all) had been acquired by 

the multinational pharmaceutical firms, acknowledging the lay public’s growing 

demand for phytomedicines in the Western World. 

The business of dietary supplement in the Western World has expanded from 

the health store to the pharmacy. Alternative medicine includes plant based products. 

Appropriate measures to ensure the quality, safety and efficacy of these 

either already exist or are being answered by greater legislative control by such 

bodies as the Food and Drug Administration of the USA and the recently created 

European Agency for the Evaluation of Medicinal Products, based in London. 

In the USA, the Dietary Supplement and Health Education Act of 1994 recognised 

the class of phytotherapeutic agents derived from medicinal and aromatic 

plants. Furthermore, under public pressure, the US Congress set up an Office of 

Alternative Medicine and this office in 1994 assisted the filing of several Investigational 

New Drug (IND) applications, required for clinical trials of some Chinese 

herbal preparations. The significance of these applications was that each Chinese 

preparation involved several plants and yet was handled as a single IND. A 

demonstration of the contribution to efficacy, of each ingredient of each plant, was 

not required. This was a major step forward towards more sensible regulations in 

regard to phytomedicines. 

My thanks are due to the staff of the Publishers who have made this series 

possible and especially to the volume editors and their chapter contributors for the 

authoritative information. 

Roland Hardman

Preface 

My interest in flower-bulb crops started in 1973, when I began working with 

Dr Alun Rees at the Glasshouse Crops Research Institute, Littlehampton, a site consigned 

to horticultural history in 1995 as a result of the all-too-familiar ‘cuts in government 

funding’. Alun had himself taken up a post at Littlehampton in 1962, when 

the (then) Agricultural Research Council started funding research on ornamental 

bulb crops. Alun succeeded in putting bulb growing and forcing on a sound scientific 

base, evidenced by the publication of his The Growth of Bulbs in 1972. 1 Fortunately 

for me, Alun’s enthusiasm for these interesting crops was infectious. 

Whilst my main research interests concerned the early forcing of bulbs, the use 

of plant growth regulators, narcissus propagation and other aspects relevant to 

the UK bulbs industry, uses of bulbs other than as ornamentals attracted my 

attention from time to time. For instance, one was aware of the eastern European 

literature, long ignored in the west, on pharmaceuticals such as galanthamine 

(galantamine) from Galanthus (snowdrops) and other genera. Only in 1995 did the 

growing of narcissus bulbs in the UK for processing for galanthamine extraction, 

and the clinical trials on the use of the compound in Alzheimer’s disease, become 

public knowledge. 2 Whilst there is a wealth both of other alkaloids in narcissus, 

and of other potential uses of galanthamine, this case in particular led to the conception 

of the present volume. Reminyl, Shire Pharmaceuticals and Janssen-Cilag’s 

Alzheimer’s disease treatment derived from narcissus, received its first European 

approval in 2000. 3,4 Subsequently, Reminyl was recommended by the National 

Centre for Clinical Excellence. 5 

The impact of Alzheimer’s disease was first brought home to me by examples of 

public figures. For example, there was ex-President Ronald Reagan’s touching 

letter to the American people, relating the start of the ‘journey that would lead 

him into the sunset of his life’. 6 Earlier, in the UK, Prime Minister Harold Wilson 

had unexpectedly retired from public life, and the cases of these two statesmen 

1 Rees, A.R. (1972) The Growth of Bulbs. Academic Press, London. 

2 Bonner, J. (1995) Flower bulbs slow brain disease. New Scientist, 145 (1964), 21. 

3 Reminyl approval lifts Shire price, The Times, 4 March 2000. 

4 New Alzheimer’s drug approved. The Pharmaceutical Journal, 265 (22 July 2000), 122. 

5 Press release 2001/002, NICE issues guidance on drugs for Alzheimer’s disease, 19 January 2001, NICE. 

6 Letter from Ronald Reagan revealing Alzheimer’s disease, for example, see http://www.law.umkc.edu/ 

faculty/projects/ttrials/hinckley/ALZHEI~1.htm

x Preface 

were poignantly contrasted in an article by Clair Woodward entitled ‘Ending the 

taboo: Reagan was right to go public over Alzheimer’s’. 7 Subsequently, I saw the 

suffering and death of my mother from Alzheimer’s disease, in 1995. My hope, 

therefore, is that this volume will prove a useful reference for those researching 

the exciting field of Amaryllidaceae alkaloids. 

I am pleased to be editor for the series of comprehensive reviews the Narcissus 

volume comprises, but am particularly gratified that much original material, and 

material accessible only with difficulty in the west, has been included. 

Notes on nomenclature 

1 Galanthamine is also referred to as galantamine; in this volume the former 

name has been used exclusively. 

2 In popular UK usage, the term ‘daffodil’ is used for ‘trumpet’ or ‘large cup’ 

types of Narcissus, and ‘narcissi’ for smaller flowered types. In the present 

volume the term Narcissus (or narcissus) has been used to cover all types, using 

the Latin at first mention or where referring to a specific cultivar or other 

taxon, otherwise using the colloquial ‘narcissus’. Where a reference is to a 

particular species or group of narcissus (e.g., Tazetta cultivars), this is stated. 

3 The taxonomy of the genus Narcissus is subject to much debate (Brian Mathew 

discusses ‘splitters’ and ‘lumpers’ in chapter 3 of this volume). While the editor 

had hoped, optimistically, to achieve uniformity of narcissus names throughout 

this volume, this did not prove practical. Except in specialist chapters, 

however, names conform as far as feasible to those given in The International 

Daffodil Register and Classified List 1998 of the Royal Horticultural Society. 


7 Woodward, C. (1995) Ending the taboo: Reagan was right to go public over Alzheimer’s. Signpost, 

1 (31), 18–19.

Acknowledgements 

I would like to thank our Series Editor, Dr Roland Hardman, for his helpfulness 

and kindly consideration during the preparation of the volume. I would also like 

to acknowledge the professionalism shown by the contributors to this book: it has 

been a pleasure working with you! As always, I appreciate the encouragement 

given by my wife Anita and daughter Esther.

Contributors 

Dr. Jaume Bastida 

Department of Natural Products 

Faculty of Pharmacy 

University of Barcelona 

08028-Barcelona 

Spain 

Dr. Jairo K. Bastos 

Universidade de São Paulo 

Faculdade de Ciências Farmacêuticas 

de Ribeirão Preto 

14040-903 Ribeirão Preto – SP 

Brazil 

Dr. Peter W. Bowers 

Department of Dermatology 

Treliske Hospital 

Truro 

Cornwall TR1 3LJ, UK 

James B. Briggs (retired) 

ADAS Consulting Ltd. 

ADAS Park Farm 

Ditton 

Aylesford 

Kent ME20 6PE, UK 

Dr. David Brown 

School of Pharmacy and 

Biomedical Sciences 

University of Portsmouth 

St Michael’s Building 

White Swan Road 

Portsmouth PO1 2DT 

UK 

Dr. V.N. Bulavka 

Slavich Company 

Mendeleev sq., 2 

Pereslavl-Zalesskiy 152025 

Russian Federation 

Dr. O.A. Cherkasov 

Department of Agriculture 

All-Russian Research Institute of 

Medicinal and Aromatic Plants (VILAR) 

Grina Str. 7 

Moscow 113628 

Russian Federation 

Dr. Carles Codina 

Department of Natural Products 



08028-Barcelona 

Spain 

Prof. Els J.M. Van Damme 

Katholieke Universiteit Leuven 

Laboratory for Phytopathology and 

Plant Protection 

Willem de Croylaan 42 

3001 Leuven 

Belgium 

Anthony C. Dweck 

Dweck Data 

8 Merrifield Road 

Ford 

Salisbury 

Wiltshire SP4 6DF 

UK


Crop and Weed Science 

Department 

Horticulture Research 

International 

Kirton, Boston 

Lincolnshire PE20 1NN 

UK 

Dr. Kornkanok Ingkaninan 

Faculty of Pharmaceutical 

Sciences 

Naresuan University 

Phitsanulok 65000 

Thailand 

Prof. Dr. Hubertus Irth 

Screentec 

Niels Bohrweg 11-13 

2333 CA Leiden 

The Netherlands 

Dr. Celia G. Julian 

Department of Dermatology 

Treliske Hospital 

Truro 

Cornwall TR1 3LJ, UK 

Dr. Mirko Kreh 

Merck KGaA 

Frankfurter Strasse 250 

D-64271 Darmstadt 

Germany 

Brian Mathew 

90 Foley Road 

Claygate 

Surrey 

KT10 ONB, UK 

Dr. Rita M. Moraes 

National Center for Natural Products 

Research 

Research Institute of Pharmaceutical 

Sciences 

School of Pharmacy 

University of Mississippi 

University 

MS 38677 

USA 

Dr. James R. Murray 

Hunter-Fleming Ltd 

Regus House 

1 Friary 

Temple Quay 

Bristol, BS1 6EA 

UK 

Contributors xiii 

Dr. N.P. Dhammika Nanayakkara 

National Center for Natural Products 

Research 

Research Institute of Pharmaceutical 

Sciences 

School of Pharmacy 

University of Mississippi 

University 

MS 38677 

USA 

Prof. Willy J. Peumans 

Katholieke Universiteit Leuven 

Laboratory for Phytopathology and 

Plant Protection 

Willem de Croylaan 42 

3001 Leuven 

Belgium 

Christian Remy 

Laboratoire Monique Remy 

Parc Industrial des Bois de 

Grasse 

F-06130 Grasse 

France 

Prof. O.N. Tolkachev 

Department of Chemistry and 

Technology of Natural 

Products 

All-Russian Research Institute of 

Medicinal and Aromatic Plants 

(VILAR) 

Grina Str. 7 

Moscow 113628 

Russian Federation

xiv Contributors 

Prof. Rob Verpoorte 

Division of Pharmacognosy 

Leiden/Amsterdam Center for 

Drug Research 

Gorlaeus Laboratories, Leiden University 

Einsteinweg 55, P.O. Box 9502 

2300 RA Leiden, The Netherlands 

Dr. Francesc Viladomat 

Department of Natural 

Products 



08028-Barcelona, 

Spain

Figures 

1.1 The effect of bulb storage at 9 or 17°C for 6–15 weeks 3 

1.2 The temperature sequence during bulb forcing, compared with 

natural temperatures 3 

1.3 Transverse section of single-nosed, flowering-size narcissus 

‘Carlton’ bulb in autumn and the generations of bulb units 

with their component parts 5 

1.4 Schematic representation of a narcissus plant showing three years’ 

development 6 

1.5 Double-nosed bulb, separation of offsets and large ‘mother bulb’ 7 

1.6 Diagrammatic representation of a large narcissus bulb showing 

the relationship between terminal and lateral bulb units 8 

1.7 The growth pattern of terminal and lateral bulb units 

from initiation to senescence, narcissus ‘Fortune’ 9 

1.8 The relationship between bulb (cluster) weight and numbers of 

bulb noses (growing points) and first-quality flowers, 

narcissus ‘Carlton’ 12 

2.1 The narcissus, a plant with a rich folklore 20 

3.1 Examples of Narcissus species of the main sections 33 

3.2 Examples of Narcissus cultivars of the main cultivar groups, 

with their classification 49 

4.1 Mean monthly weather data 66 

4.2 Typical narcissus fields of south Lincolnshire and old 

daffodil beds on St. Marys, Isles of Scilly 67 

4.3 Modern front-loading hot-water treatment tanks 74 

4.4 A typical bulb planting machine feeding bulbs from 

a hopper into two ridges 80 

4.5 Narcissus bulb yields for one- and two-year-down growing 81 

4.6 Effect of planting density, bulb grade and crop 

duration on profitability. Data for narcissus ‘Fortune’ 

grown in ridges 83

xvi Figures/Plates 

4.7 Changes in plant dry weight and leaf area index during the 

growing season. Data for narcissus ‘Golden Harvest’ 92 

4.8 Large unmanned bulb lifter discharging into bulk trailer 93 

4.9 Letter box drying wall for bulbs in bulk bins 95 

4.10 Narcissus ‘chips’ with bulblets at the end of incubation 107 

6.1 Sceletium type alkaloid mesembrenone isolated from N. pallidulus 142 

6.2 Narcissus alkaloid types 143 

6.3 Biosynthetic pathway to norbelladine 164 

6.4 Oxidative phenyl-phenyl coupling in Amaryllidaceae alkaloids 165 

6.5 Alkaloids proceeding from an ortho-para′ coupling 165 

6.6 Biosynthesis of lycorine with inversion of the configuration 166 

6.7 Conversion of galanthine to narcissidine via epoxide 166 

6.8 Conversion of norpluviine to homolycorine type alkaloids 167 

6.9 Alkaloids proceeding from a para-para′ coupling 168 

6.10 Biosynthesis of pretazettine 169 

6.11 Proposed biosynthetic pathways to haemanthamine 

and montanine 170 

6.12 Biosynthesis of galanthamine 171 

6.13 Mass fragmentation pattern of lycorine 180 

6.14 Mass fragmentation pattern of homolycorine 181 

6.15 Mass fragmentation pattern of haemanthamine 182 

6.16 Mass fragmentation pattern of tazettine and criwelline 183 

6.17 Mass fragmentation pattern of montanine 183 

6.18 Mass fragmentation pattern of galanthamine 184 

7.1 Alkaloid content in friable and meristematic callus of 

seed-derived explants 217 

7.2 Alkaloid content in shoot clusters grown in medium with 

BA or kinetin 218 

7.3 Levels of alkaloids released by root clusters to the culture 

medium. Accumulation of alkaloids in tissue and 

liquid medium 219 

7.4 Alkaloid content in shoots and their respective twin-scales 

according to their position in the bulb 220 

7.5 Levels of alkaloids released by shoot-clumps treated with BA 

into the culture medium 222 

7.6 Accumulation of alkaloids in both tissue (shoot-clumps) 

and liquid medium in the experiment with BA 223 

7.7 Levels of alkaloids released by shoot-clumps treated with 

kinetin into the culture medium 224

Figures/Plates xvii 

7.8 Accumulation of alkaloids in both tissue (shoot-clumps) and 

liquid medium in the experiment with kinetin 225 

7.9 Levels of alkaloids released by bulblets treated with paclobutrazol 

into the culture medium 227 

7.10 Accumulation of alkaloids in both tissue bulblets and liquid 

medium in the experiment with paclobutrazol 228 

7.11 Levels of alkaloids released by shoot-clumps treated with 

paclobutrazol into the culture medium 229 

7.12 Accumulation of alkaloids in both tissue (shoot-clumps) 

and liquid medium in the experiment with paclobutrazol 230 

7.13 Alkaloid production in bulb-derived shoot-clumps grown 

under different sucrose concentrations 233 

7.14 Alkaloid production in seed-derived shoot-clumps grown 

under different sucrose concentrations 234 

7.15 Alkaloid production by plantlets grown in 250ml flasks 236 

7.16 Alkaloid production by plantlets grown in 500ml flasks 237 

7.17 Comparison between the total production of alkaloids 

by plantlets grown in two different flask sizes 238 

8.1 Formulae of galanthamine, lycorine and fortucine 243 

9.1 Narcissus ‘Carlton’ 259 

9.2 Galanthamine content of bulbs of Narcissus ‘Carlton’ 

over one year 260 

9.3 Galanthamine content of aerial parts of Narcissus ‘Carlton’ 

over one year 262 

9.4 Galanthamine content of the different plant organs of Narcissus 

‘Carlton’ on a fresh weight basis 263 

9.5 HPLC chromatograms of the alkaloids obtained from 

different parts of flowering Narcissus ‘Carlton’ 264 

9.6 Galanthamine content in leaf and bulb of Narcissus ‘Carlton’ 

following the application of different fertilisers 265 

9.7 HPLC chromatograms of the alkaloid fractions 266 

10.1 The effect of developmental stages of Narcissus ‘Inglescombe’ 

on galanthamine content during the 1994 growing season 278 

11.1 Chemical structures of Amaryllidaceae alkaloids with 

important biological activities 286 

12.1 The equilibrium process between narwedine hydrohydienone 

and its enantiomers. Structure of norbelladine and 

N,O-dimethylnorbelladine 305 

12.2 Synthesis of narwedine via N,O-dimethylnorbelladine 307 

12.3 Synthesis of narwedine via palladium-containing intermediate 308 

12.4 Synthesis of galanthamine via belladine-type benzamide 309

xviii Figures/Plates 

12.5 Synthesis of 4-benzyloxyphenylacetic acid and 2-bromo- 

4-methoxy-5-benzyloxy-benzaldehyde 310 

12.6 Synthesis of galanthamine via N-norgalanthamine and via 

belladine-type phenylacetamide 312 

12.7 Synthesis of galanthamine via non-brominated belladine-type 

benzamide 313 

12.8 Synthesis of galanthamine via photochemical substitution of 

bromine in belladine-type benzamide 314 

12.9 4-Methoxybenzyl-protected intermediates for the synthesis of 

galanthamine. Microbiological reduction of bromo-narwedinone 315 

12.10 Synthesis of enantiomeric galanthamine 316 

12.11 Synthesis of galanthamine via N-formylnorbelladine derivative 317 

12.12 Synthesis of phenylethylamine intermediates and galanthamine 319 

12.13 Synthesis of narwedine-type enones and dienones by 

electrochemical oxidation of belladine-type amides 321 

12.14 Synthesis of N,O-didemethyl-N-trifluoroacetylnarwedine and 

N-demethyl-N-trifluoroacetyl-narwedine 323 

12.15 Synthesis of narwedine, galanthamine, lycoramine and sanguinine. 

Reactions of trimethylsilanyl-substituted narwedine-type dienone 324 

12.16 Oxidative cyclisation reactions of trimethylsilanyl-substituted 

norbelladine-type trifluoroacetamides to benzazepine 

spirodienones 326 

13.1 Chemical structures for galanthamine, morphine, 

1-adamantyl demethyl galanthamine and pretazettine 333 

13.2 Galanthamine metabolic pathway 338 

14.1 Schematic representation of the action of galanthamine 357 

15.1 Percentage of inhibition effect of methanol extracts from some 

narcissus cultivars at the concentration of 0.1mg/ml measured 

with the microplate assay 372 

15.2 Scheme of the on-line HPLC-UV-MS-bioactivity detection 

for acetylcholinesterase inhibitors 373 

15.3 The spectra and the chromatograms obtained after injection 

of the fraction from narcissus ‘Carlton’ extract into the on-line 

HPLC-UV-MS-bioactivity detection system 374 

15.4 The spectra and the chromatograms obtained after injection 

of the fraction from narcissus ‘Sir Winston Churchill’ extract 

into the on-line HPLC-UV-MS-bioactivity detection system 375 

15.5 Structure of ungiminorine and galanthamine 375 

16.1 Schematic representation of the biosynthesis, processing and 

topogenesis of the narcissus lectin (NPA) 384 

16.2 Deduced amino acid sequence of the narcissus lectin precursor 

encoded by cDNA clone LECNPA1. Internal sequence similarity

Figures/Plates 

of different segments of the mature narcissus lectin sequence of 

xix 

109 amino acids 385 

16.3 Three-dimensional structure of the narcissus lectin monomer 385 

17.1 Narcissus (Narcissus poeticus) can be seen like white sheets 

in pastures 393 

17.2 Flower collecting implements 394 

17.3 Narcissus flowers spread out to dry, and a drum holding 

20kg of concrete 396 

17.4 Scheme for the production of narcissus absolute 396 

18.1 Daffodil picking: snapping off the flower stem 400 

18.2 Granulomatous rash on the wrist 400 

18.3 Daffodil picking: gathering up the bunches 401 

18.4 Secondary facial rash 402 

18.5 Polymorphic calcium oxalate crystals 404 

Plates 

Colour plates appear between pages 140 and 141 

1 Examples of Narcissus species of the main sections 

2 Examples of Narcissus cultivars of the main cultivar groups, 

with their classification 

3 Granulomatous rash on the wrist 

4 Secondary facial rash 

5 Polymorphic calcium oxalate crystals

1 The biology of Narcissus 


INTRODUCTION 

The genus Narcissus L. belongs to the Monocotyledon family Amaryllidaceae, to 

which it contributes some 80 species to its total of about 850 species in 60 genera 

(Meerow and Snijman, 1998). The taxonomy of Narcissus is difficult because of the 

ease with which hybridisation occurs naturally, accompanied by extensive cultivation, 

breeding, selection, escape and naturalisation (Webb, 1980; see Chapter 3, 

this volume). The genus is distinguished from other Amaryllids by the presence of 

a perigonal corona structure (‘paraperigone’) forming a ring (‘cup’) or tube 

(‘trumpet’) (Dahlgren et al., 1985). Unlike other genera of the family, Narcissus has 

a mainly Mediterranean distribution, with a centre of diversity in the Iberian 

Peninsula, and the genus also occurs in south-western France, northern Africa and 

eastwards to Greece, while Narcissus tazetta is found not only in Spain and North 

Africa but in a narrow band to China and Japan (Grey-Wilson and Mathew, 1981). 

The eastwards distribution of N. tazetta may represent transfer along an ancient 

trade route, illustrating the long human interest in the genus as an ornamental 

plant, leading to its importance in commercial horticulture today (see Chapter 4, 

this volume). 

The survival of a number of Narcissus species has been threatened by past overcollection 

and habitat destruction, not only in Spain and Portugal but also in 

Morocco, Turkey and Belgium (Oldfield, 1989; Koopowitz and Kaye, 1990). The 

‘Red List’ currently gives three Narcissus as ‘endangered’, five as ‘vulnerable’ and 

six as ‘rare’ (WCMC, 1999). In the light of the environmentalist concerns in the 

1990’s, the collection of wild bulbs has been addressed by the industry. However, 

there is a need to maintain vigilance in the conservation of wild species and of their 

many variants, to establish genetic collections for future breeding programmes, 

and to develop sustainable production systems for their utilisation in commercial 

horticulture. 

Hybridisation has resulted in commercial narcissus cultivars that are in most 

cases larger and more robust than their wild parents. Trumpet cultivars with 

coloured perianth and corona originated from N. pseudonarcissus and its varieties, 

and trumpet cultivars with white perianth and coloured corona from N. pseudonarcissus 

ssp. bicolor. Large-cupped cultivars were the result of crosses between 

N. pseudonarcissus and N. poeticus, back-crossed with N. poeticus to yield the smallcupped 

cultivars. Multiheaded cultivars (the ‘Poetaz’ group) comprise mainly 

hybrids of N. poeticus and N. tazetta (Doorenbos, 1954).

2 G.R. Hanks 

Information about the commercial horticulture of narcissus can be found in 

Rees (1972, 1985b, 1992) and Hanks (1993). Texts on the genus include Bowles 

(1934), Jefferson-Brown (1969, 1991), Blanchard (1990) and Wells (1989). 

THE GROWTH CYCLE UNDER NATURAL CONDITIONS 

The native habitats of Narcissus species are very varied, and include grassland, scrub, 

woods, river banks and rocky crevices, in both lowland and mountain sites (Webb, 

1980), and the ecology of natural populations of wild daffodil (N. pseudonarcissus) has 

been intensively studied (Caldwell and Wallace, 1955; Barkham, 1980a,b, 1992; 

Barkham and Hance, 1982). The bulk of Narcissus species are synanthous and 

spring-flowering. Shortly after flowering rapid leaf senescence occurs, followed by 

a summer underground (or ‘dormant’) period that allows the bulb to conserve 

moisture and avoid predators (the alkaloids which are largely the subject of the 

present volume may give further disincentive to predation). Although the term 

‘dormancy’ is used, this refers mainly to the lack of any obvious external growth, 

for there is little physiological dormancy because, once the leaves and roots have 

died down, there is intense activity of primordia within the bulb (Kamerbeek et al., 

1970; Rees, 1971, 1972). There is a requirement for a cold period before normal 

growth resumes in the spring, an arrangement that avoids most damage due to 

frosts in winter. The cold requirement, however, is not particularly long nor cold, 

so that in climates like the UK or the Netherlands it is easily satisfied by normal 

winters. The flowering date is then dependent on spring temperatures being sufficiently 

high for growth, the resultant variations in flowering date being described 

by Rees and Hanks (1996). The cold requirement is not an obligate one for stem 

extension, as stem growth will proceed even without a cold treatment, albeit slowly 

and with a gradual loss of flowers due to bud abortion or other causes (Rees and 

Hanks, 1984). The effect of the cold period is to produce rapid, synchronous stem 

extension and progress to anthesis (Figure 1.1). In nature, this occurs at a time 

when competition for pollinator insects is low and growth is relatively unhindered 

by shading or other competition from grasses or deciduous trees (Caldwell and 

Wallace, 1955; Shmida and Dafni, 1990). 

The growth cycle just described has been utilised in commercial horticulture, 

being manipulated by controlled-temperature storage (Figure 1.2) to obtain ‘forced’ 

flowers in glasshouses over an extended season (ADAS, 1985; De Hertogh, 1989; 

Anon., 1998). Geophytic plants such as bulbs and corms lend themselves to horticultural 

usage, since their storage organs can be conveniently treated (whether by 

pesticides or environmental treatments), transported and traded, yet can be 

brought into flower in a relatively short time. The rapid growth and flowering of 

spring bulbs is dependent on the conversion of insoluble reserves, such as starch, 

into readily translocatable soluble sugars. In order to understand these processes 

and develop improved methods of flower forcing, carbohydrate metabolism and 

the related hormone-mediated processes have been investigated in flower-bulb 

crops, although less so in the case of narcissus than in species such as tulip and iris, 

perhaps because of the presence in narcissus of mucilaginous sap, which interferes 

with chemical extraction and separation. Carbohydrate metabolism in narcissus 

has been studied by Grainger (1941), Thomas et al. (1995) and Ruamrungsri et al.

Time to anthesis (days) 

250 

200 

150 

100 

50 

0 

9°C (time) 17°C (time) 9°C (length) 17°C (length) 

6 9 12 15 

Duration of storage (weeks) 

Figure 1.1 The effect of bulb storage at 9 or 17 °C for 6–15 weeks on (left axis) the time 

to anthesis (days in a glasshouse at 16 °C) and (right axis) stem length at 

anthesis for narcissus ‘Fortune’ (data from Rees and Hanks, 1984). 

Bulb drying 

Bulb storage 17 °C 

Cold storage 9 °C 

Artificial temperatures 

Holding temperature 0 –5 °C 

Glasshouse temperature 15 °C 

Natural temperatures 

Summer Autumn 

Winter 

Spring 

Figure 1.2 The temperature sequence during bulb forcing, compared with natural 

temperatures (after A.R. Rees, personal communication). 

400 

300 

200 

100 

0 

Stem length at anthesis (mm)

4 G.R. Hanks 

(1999), and the polysaccharides by Balbaa et al. (1980) and Rakhimov and Zhauynbaeva 

(1997). The auxins of narcissus have been studied by Edelbluth and Kaldewey 

(1976), gibberellins by Aung et al. (1969), cytokinins by van Staden (1978) and 

Belynskaya et al. (1990) and ethylene by Staby and De Hertogh (1970). 

Cultivars derived from the N. tazetta group are unusual in that, while still 

‘summer dormant’, they have no cold requirement and growth and anthesis can 

occur before winter if all other conditions are favourable (although cold treatments 

can increase stem length and speed anthesis; Roh and Lee, 1981; Rees and Hanks, 

unpublished data in Hanks, 1993). Tazetta narcissus, unlike most narcissus, 

respond to ethylene, which promotes flowering (Imanishi, 1997). These characteristics 

have enabled horticulturists to exploit Tazettas for flower production over a 

long season. 

A few species – N. elegans, N. serotinus, N. viridiflorus and N. humilis – are autumnflowering 

and generally hysteranthous. Studies on N. tazetta and other geophytes 

suggest that the syanthous-hysteranthous habit is facultative, with hysteranthy 

tending to be expressed in xeric habitats (Evenari and Gutterman, 1985; Halevy, 

1990). The autumn-flowering species have not yet been exploited horticulturally, 

probably because their flowers are small or insignificant, although they clearly 

have potential for producing new types of commercial cultivars (Koopowitz and 

Kaye, 1990). 

MORPHOLOGY AND DEVELOPMENT 

The flowering plant 

Detailed reports of the morphology and development of the narcissus plant have 

been given by Huisman and Hartsema (1933), Chan (1952), Okada and Miwa 

(1958) and Rees (1969, 1972), from which the following description has largely 

been compiled. 

Bulb structure and bulb unit development 

The ‘dormant’ narcissus bulb in autumn consists of a more-or-less disc-shaped 

stem plate (base or basal plate) bearing adventitious roots below and storage 

organs (bulb scales) surrounding a bud above. The bulb ‘scales’ consist of true 

scales, which are almost entirely within the bulb and serve a purely storage function, 

and the bases of foliage leaves; after anthesis the base of the flower stalk becomes 

flattened and is also scale-like in function. Leaf bases can be distinguished from 

bulb scales because the former have a thicker tip and a scar where the leaf lamina 

became detached. In the ‘model’ case of a narcissus bulb with a single, terminal 

growing point (a ‘single nosed round’ bulb), a transverse section of the bulb in 

autumn shows a series of concentric ‘scales’ surrounding the old flower stalk base 

and a terminal bud (Figure 1.3). The exception to the concentric nature of the 

‘scales’ is that the inner leaf, which subtends the flower, has a semi-circular base 

with keeled margins only partly enclosing the flower stalk. The terminal bud 

consists of bulb scales surrounding leaves and a flower. In addition to the terminal 

bud, there is also a lateral bud.

The biology of Narcissus 5 

Figure 1.3 Left: transverse section of single-nosed, flowering-size narcissus ‘Carlton’ bulb 

in autumn. Right: diagram showing the generations of bulb units with their 

component parts. Shaded areas: the terminal (above) and lateral (below) units 

with next spring’s leaves and flower (individual bulb scales, leaves and flower 

not shown). The unit that bore last spring’s leaves and flower comprises the 

flattened remains of the stem (cross-hatched), three leaf bases (the innermost 

semi-sheathing) (stippled) and two bulb scales (unshaded). Beyond these scales 

are the remains of bulb scales and leaf bases of previous generations of bulb 

units, which are eventually shed as dry tunic (broken lines). (After Hanks (1993); 

reprinted from The Physiology of Flower Bulbs, ©1993, page 466, with the permission 

of Elsevier Science.) 

The narcissus is a perennial branching system, and Rees (1969, 1972, 1987) 

used the term ‘bulb unit’ to describe each annual increment of growth, thereby 

distinguishing these structures from shorter lived entities such as the bulbs of 

tulip, where new bulbs (daughter bulbs) become separate entities each year. In narcissus, 

the growing point produces a new bud with bulb scales and leaves each 

year. This bud grows through its first year, and (if large enough) initiates a flower 

in its second year, its leaves and flower emerging in the next spring. Thereafter its 

‘scales’ persist for perhaps two more years, so that the bulb unit has an overall lifespan 

of about 4 years. Since new buds are initiated in the centre of the bulb, the 

bulb ‘scales’ are gradually displaced outwards by the continued annual production 

of new bulb units within. In the year that a bulb unit reaches anthesis, its ‘scales’ 

(swollen with reserves) make up the bulk of the fresh weight of the ‘bulb’; subsequently 

its ‘scales’ become depleted until they form the dry tunic (skin) of the bulb, 

and they are eventually lost through displacement from within or by abrasion (the 

latter accentuated by commercial bulb handling). Although bulb units may be 

called ‘mother bulbs’ in the year they reach anthesis and ‘daughter’ and ‘granddaughter 

bulbs’ previously, better terms might be pre-floral, floral and post-floral 

bulb units. What are termed ‘bulbs’ in common usage might be better called ‘compound 

bulb units’ or, more simply, ‘bulb clusters’. A useful diagram showing 

narcissus development was presented by Alkema and van Leeuwen (1978) and is 

reproduced in Figure 1.4.

3 

2 

4 

1 

1 

3 

2 

4 

5 

Flowering in 1976 

4 

6 

4 

8 

2 

6 


2 


1 

3 

5 

1 

5 

3 

4 

1 

3 

5 

7 

2 

7 

2 

4 

1 

3 

Cluster August 1976 


Base of leaf blade 

Base of scale leaf 

Depleted scale 

Bud giving rise to a 

flowering shoot 

Bud giving rise to a 

non-flowering shoot 


Figure 1.4 Schematic representation of a narcissus plant showing three years’ development 

(redrawn after Alkema and van Leeuwen (1978), with permission from the Bulb 

Research Centre, Lisse, The Netherlands).


Figure 1.5 Centre: double-nosed bulb, narcissus ‘Carlton’. Right: separation of offsets. 

Left: offset (top) and large ‘mother bulb’ (below). (Photograph: Horticulture 

Research International.) 

As narcissus bulbs are branching systems, lateral bulb units are initiated as well 

as the terminal, replacement units. The presence of lateral as well as terminal bulb 

units, and the gradual separation from the cluster of bulb units derived from laterals 

(‘offsets’), results in a variety of bulb shapes and sizes, from round single-nosed 

bulbs to double- and multi-nosed bulbs (the latter also termed ‘mother bulbs’), 

together with smaller attached or detached, usually non-flowering, offsets (Figure 1.5). 

The life-span of individual branching systems is not known for narcissus cultivars, 

although Barkham (1980a) recorded half-lives of adult N. pseudonarcissus of 12–18 

years, and Koopowitz (1986) suggested that, judging from other Amaryllids, they 

are likely to have a high longevity. 

Terminal bulb units are initiated alongside the flower initial of the previous bulb 

unit, in the axil of the second leaf from its centre, at or shortly after floral initiation 

in May (Rees, 1969). Lateral bulb units are initiated, usually in the axil of the third 

leaf from the centre, in the following December. Supernumerary (lateral) bulb 

units may also be initiated in the same year as regular laterals or a year later, in the 

axil of a bulb scale (usually the innermost scale) or alongside the regular lateral. 

The classification of bulb units was elegantly described by Rees (1969) (Figure 1.6, 

Table 1.1). Each terminal bulb unit is replaced by two ‘daughter’ units, terminal 

and lateral, while lateral units rarely contain a lateral unit. This gives a slow increase 

in bulb unit numbers, increasing in a Fibonacci series 1, 1, 2, 3, 5, 8, 13..., an 

increase tending to 1.6-fold per annum, producing a population made up of 38% 

lateral units. In commercial bulb growing, this progression is prevented by the 

grading-out of saleable bulbs, and, possibly, by the suppression of laterals under 

sub-optimal conditions. In the earlier study by Okada and Miwa (1958) on a

8 G.R. Hanks 

Figure 1.6 Diagrammatic representation of a large narcissus bulb showing the relationship 

between terminal (T) and lateral (L) bulb units. Each circle represents 

one bulb unit (or annual increment of growth in the branching system), the 

small black ones being those of the current season, labelled with a threeletter 

code. Successively older units are one size larger, and coded with one 

or two letters, respectively. (After Rees (1969), with permission of Academic 

Press Ltd.) 

different cultivar, bulbs produced an average of 2.2 new bulb units annually, and 

in this case 19% were formed in the axils of bulb scales. 

After terminal bulb units are initiated in May, they grow rapidly for three 

to four months and then more slowly through autumn and winter (Rees, 1969). 

Periods of alternating rapid spring-summer and slow autumn-winter growth continue, 

and the bulb unit reaches a peak dry weight in the May two years after its initiation, 

shortly after anthesis. There is some weight loss the following spring, corresponding 

to the rapid growth of the next generation of bulb units, after which weight is 

regained and maintained until August, following which the bulb unit becomes 

senescent and dies. Lateral bulb units show less distinct alternations of growth 

rates, as they are initiated later than terminals (Figure 1.7). Periods of rapid 

growth of bulb units begin in February, reserves being translocated from older 

bulb units. No new parts are being produced at this time, so this is a true bulbing


Table 1.1 The production of terminal (T) and lateral (L) bulb units in Narcissus a 

Year 1 2 3 4 5 6 7 

Bulb unit types T T T TT TTT TTTT TTTTT 

L TL TTL TTTL TTTTL 

LT TLT TTLT TTTLT 

LTT TLTT TTLTT 

LTL TLTL TTLTL 

LTTT TLTTT 

LTTL TLTTL 

LTLT TLTLT 

LTTTT 

LTTTL 

LTTLT 

LTLTT 

LTLTL 

No. of bulb units 1 1 2 3 5 8 13 

Increase over previous year – 1 2 1.5 1.7 1.6 1.6 

Note 

a After Rees (1969). 

Figure 1.7 The growth pattern of terminal (T) and lateral (L) bulb units from initiation 

to senescence, narcissus ‘Fortune’. The times of ‘scale’ initiation, floral initiation 

and anthesis are indicated by 1, 2 and 3. (Modified from Rees (1969), 

with permission of Academic Press Ltd.) 

effect probably controlled by temperature; attempts to show a photoperiodic effect 

were not successful (Rees, 1972). 

In both terminal and lateral bulb units the full complement of bulb scales and 

leaves is initiated during its first period of active growth, and the apices then 

become inactive until the initiation of the flower at the end of the second period 

of rapid growth of the terminal bulb unit, a year after the initiation of that unit

10 G.R. Hanks 

(Rees, 1969). When floral initiation does not occur, there may be renewed initiation 

of leaves, resulting in non-flowering bulb units with high numbers of leaves. If floral 

initiation occurs in lateral bulb units, it is after their period of rapid growth, and 

several months after floral initiation in the terminal bulb unit. In contrast to terminal 

bulb units, laterals have fewer parts, are lighter in weight, have less tendency to 

flower, and show almost complete suppression of further lateral units. Rees (1972) 

suggested that the differences between the two types of bulb unit were due to the 

later initiation of lateral bulb units, and because lateral units are subject to apical 

dominance by the terminal units and develop only once the terminal has become 

floral, losing its dominance. Large number of new bulb units are formed when the 

shoot apices are damaged by pests, disease or high temperature. This suggests that 

there is a capacity for regeneration at the base of the scales which is not normally 

expressed because of apical dominance (Rees, 1972), an observation confirmed by 

the effectiveness of propagation techniques such as twin-scaling (see Chapter 4, 

this volume). 

The shoot apex and primordia initiation 

The anatomy of the narcissus stem apex was described by Denne (1959). There is 

no conclusive evidence as to whether the flower of narcissus is terminal as a result 

of sympodial branching (in which case an axillary bud becomes the new growing 

point) or axillary as a result of monopodial branching (in which case the main axis 

continues vegetative growth) (Huisman and Hartsema, 1933). Rees (1972) summarised 

the arguments, favouring the sympodial view partly on the basis of the 

appearance of serial sections through the initiating apex. The floral primordium 

comes to dominate the apex in any case, with the vegetative apex remaining 

quiescent for some time. 

Denne (1960) described the comparative development of bulb scales and leaves. 

The factors controlling the transition between scale and leaf production on the 

apex are not known, and there is a period of apical inactivity between the formation 

of the two types of primordia (Rees, 1972). While apex size was shown to be 

related to the width of leaves formed in terminal or lateral bulb units, it did not 

appear related to whether scales or leaves were formed first (Denne, 1960). 

Whereas in tulips, there are examples of laminae forming on bulb scales (after 

treatments that kill the foliage leaves), no such instances were noted in narcissus 

(Rees, 1972). The anatomy of bulb scales and leaves is similar up to a length of 

1 mm: thereafter, the whole division is restricted to the base of bulb scales, 

whereas, in the leaf, cell division occurs in the basal sheath and in an intercalary 

region of the lamina, the latter continuing until the lamina is around half its final 

length (Okada and Miwa, 1958; Denne, 1960). The anatomy of scales and leaf 

bases is similar (Chan, 1952). 

Observations have been made on the relative number of bulb scales and leaves 

in bulb units. Rees (1969) obtained data for cultivars ‘King Alfred’ and ‘Fortune’, 

but the commonest combination in the former (3 + 3) occurred only once in 594 

units of ‘Fortune’ examined, suggesting there were varietal differences in the ratio 

of the two structures. Higher complements of scales and leaves occurred in terminal 

units than in laterals, and on terminal units in laterals than on its laterals, perhaps 

related to the earlier initiation of terminal units. Of terminal units, almost all


flowered, and those that did not produced four or more leaves; in contrast, few 

laterals flowered, usually if there were three or four leaves. 

There are no experimental data on the factors which trigger floral initiation 

(Rees, 1972). Experimentation is difficult as initiation takes place before bulb lifting, 

and would be complicated by the perennial habit and, hence, possibly the effects 

of earlier years. However, bulb units appear to reach a critical weight before they 

are likely to contain flowers. In many bulbous ornamentals, flower initiation occurs 

at a fixed time in the normal annual cycle of development, and often, for example 

in tulip and iris, after a minimum number of leaves have been initiated, perhaps 

related to apex size (Rees, 1985a). A similar situation may exist in narcissus, as 

there appears to be some relationship between leaf numbers and flowering, albeit 

different in terminal and lateral bulb units (see above). Rees (1986) determined 

the critical weight for flowering, based on bulb unit weights: using bulbs dissected 

in August, the critical weight was about 1.15 g, although there was some overlap in 

the weight distribution of flowering and non-flowering units. The critical weight 

for flower initiation of bulb units, and hence critical weight for clusters, may vary 

from year to year (Dickey, 1940; Rees, 1986), with growing conditions (Roh et al., 

1978; Kim and Lee, 1982), and between bulbs propagated by chipping and ‘ordinary’ 

bulbs (ADAS, 1987). 

The number of flowers per bulb or per weight of bulbs is important for commercial 

bulb producers. Rees (1986) examined the relationships between the 

number of bulbs and flowers per tonne of bulbs, the number of bulb units per 

bulb, and the number of flowers per bulb unit. Variations in flower yields might 

arise for a number of reasons, such as a large number of bulb units being below 

critical size, the occurrence of a few very large bulb units, or variations in the critical 

size itself. Over four years, the number of flowers obtained varied from 27.5 to 

32.9 thousands/tonne in ‘Carlton’ and from 21.0 to 31.8 in ‘Golden Harvest’. A 

major cause of variation in flower yield was the number of bulb units per unit 

weight in larger bulbs, which was only partly compensated by an increase in the 

number of bulb units per bulb, although the number of bulb units per tonne was a 

relatively consistent statistic. The number of flowers per bulb unit varied between 

cultivars and years, and the critical bulb unit weight for flowering varied from year 

to year. To obtain high numbers of flowers per tonne of bulbs, mean bulb unit 

weight should just exceed the critical weight, and cluster weights should be multiples 

of the critical weight, as illustrated by the analyses of Alkema and van Leeuwen 

(1978) and Kruyer (1981) (Figure 1.8). At present the factors controlling these 

responses are not known, and flower yields can be manipulated only crudely 

through changing the grade of bulbs planted, planting density and the duration of 

the crop (growing the crop for one or more years). 

Experiments carried out by Gerritsen and van der Kloot (1936) and Hartsema 

(1961), involving lifting bulbs early (around or before floral initiation) and excising 

leaves, suggested that, under normal conditions, the presence of green leaves was 

essential for floral initiation. However, in bulbs cold-stored for 6 months, new 

flowers were sometimes initiated in the lateral bulb units in the absence of green 

leaves (Hartsema and Blaauw, 1935). This suggested that the effects of light are 

not essential for flower initiation to take place (Hartsema, 1961). Although the 

effects of temperature on floral initiation are largely unknown, there is much 

information on the effects of temperature on flower development and production

12 G.R. Hanks 

Figure 1.8 The relationship between bulb (cluster) weight and numbers of bulb noses 

(growing points) and first-quality flowers, narcissus ‘Carlton’. (Modified 

after Kruyer (1981), with permission of the publishers of Bloembollencultuur.) 

in relation to commercial flower forcing, where rapid and carefully timed stem 

extension and anthesis are necessary (e.g., see Rees, 1972; Hanks, 1993). Hartsema 

(1961) reported that the optimum temperature for flower formation was initially 

20 °C, falling gradually to 13 °C as development progressed. Low temperatures 

(9–15 °C) immediately after lifting favoured early anthesis. For optimal growth 

subsequently, 11°C was required until there was 3 cm of growth, then 17 °C until 

6 cm of growth, then 20 °C. In N. tazetta types, the optimum temperature for flower 

initiation was 25–30 °C (Yahel and Sandler, 1986; Koike et al., 1994). Smoke and 

ethylene treatments induce flowering in bulbs of Tazetta narcissus below flowering 

size (Imanishi, 1997), but do not affect the flowering of standard daffodil cultivars 

(Tompsett, 1985). 

Above-ground parts 

The fully expanded foliage leaf consists of a basal sheath and a lamina, the prolongation 

of one side of the basal part. The lamina is ribbon-like, boat-shaped in 

transverse section (owing to an abaxial, mid-line ridge), with parallel venation and 

isobilateral symmetry, except in species such as N. juncifolius and N. jonquilla where 

the leaf is semi-cylindrical or rush-like (Chan, 1952). The mature bulb scale has a 

semi-transparent prolongation forming a short-lived sheath that extends above 

the soil, enclosing and supporting the foliage leaves early in their growth. Each 

side of the lamina has a similar anatomy, but at the base of the lamina elongated 

cells on the abaxial side contain raphides of calcium oxalate (Chan, 1952). Chan 

(1952) examined more than 100 Narcissus cultivars, of which 70% had two to four 

leaves, but some cultivars were consistently leafier, with seven to eight leaves.


The flower stalk is a leafless single internode (or scape), which contains abundant 

starch grains and raphides of calcium oxalate (Chan, 1952). The scape is topped 

by a single flower or a cyme, the receptacle of which bends to bring the flower into 

a horizontal position for anthesis and straightens after fertilisation to hold the 

capsule upright, a tropic response to gravity accentuated by light. Flowers also 

turn towards the light when growing in clumps (Rees, 1988). After flowers have 

been picked, the lower part of the stem that is left behind is capable of considerable 

growth from its basal meristem. 

The flower bud is enclosed in a protective sheath or spathe, which is green during 

bud development and dries and splits before anthesis. Flower development in 

narcissus was described and reviewed by Huisman and Hartsema (1933) and Chan 

(1952). The flower consists of two whorls of three perianth segments (or tepals) 

which arise from a hypanthial tube, two whorls of three anthers, and a tricarpellate 

inferior ovary with two rows of anatropous ovules in each of three loculi. The 

corona (paracorolla, trumpet or cup), characteristic of the genus, is conspicuous 

between the perianth segments and the anthers (Webb, 1980); it is regarded as 

either a prolongation of the receptacle or as a distinct structure (Huisman and 

Hartsema, 1933; Guédès, 1966). The floral parts are formed in a spiral sequence 

from the perianth inwards to the gynoecium, although for horticultural purposes 

the completion of flower initiation (important as a key stage in beginning cold 

storage or hot-water treatment) is regarded as when the corona initial can be 

clearly seen on dissection, following the initiation of the gynoecium (Huisman and 

Hartsema, 1933; Cremer et al., 1974). There are many varieties with double flowers, 

the structure of which varies in complexity (Reynolds and Tampion, 1983). Gross 

flower abnormalities (such as two flowers from the same apex) are rare (Chan, 

1952), although physiological disorders may occur in flower development (Rees, 

1972). The floral biology of N. pseudonarcissus has been described in detail by Caldwell 

and Wallace (1955). The corona and perianth tube lead to nectaries between 

the staminal filaments, the flowers being pollinated by bumble bees (Bombus spp.). 

Pollination is often poor due to a lack of insects in the relatively cool conditions, 

and little pollination occurs in the absence of insects. Pollination of N. longispathus 

in Spain was studied by Herrera (1995); the temperature inside the flower was up 

to 8 °C above ambient temperatures, raising the temperature of pollinating bees 

sufficient for flight. Di- and polymorphism with respect to style length has been 

reported for N. tazetta and N. triandrus, respectively (Arroyo and Dafni, 1995; Barrett 

et al., 1997). The fruits are ovoid green pods which grow to full size in about two 

weeks and dehisce a few weeks later. The seeds are black and round and are 

distributed over a restricted area; seeds of commercial narcissus cultivars were 

described by Chan (1952). 

Roots 

The roots of narcissus are adventitious, unbranched, with a prominent root cap, 

and are generally considered to have no root hairs, although a few poorly 

developed root hairs can occur in some circumstances (Chan, 1952; Rees, 1972; 

Kawa and De Hertogh, 1992). However, Chilvers and Daft (1981) found root hairs 

in all cultivars examined, while Wilson and Peterson (1982) described the roots of 

N. lobularis as initially glabrous and later with hairs. Price (1977) examined the

14 G.R. Hanks 

root system of commercial narcissus, reporting a maximum of 112 functional roots 

for a bulb of 12–14 cm circumference, with 77% of root weight occurring in a 

20 cm-deep layer beneath the bulb, indicating little depth or lateral spread. Some 

roots, shorter and more persistent than the rest, are contractile (Chen, 1969; Price, 

1977). Contraction occurs near the base plate, shortening the root by 7–8mm over a 

four week period in spring (Chen, 1969), and its relationship to cell wall structure 

was investigated by Wilson and Anderson (1979). Contraction in N. tazetta was 

stimulated by illumination of the lower part of the leaf laminae (Putz, 1996). The 

presence of endotrophic mycorrhizae in narcissus roots has been described 

(Kelley, 1950; Chan, 1952; Chilvers and Daft, 1981; Iqbal and Firdaus, 1986a,b). 

Roots rapidly became infected with mycorrhizae soon after planting, infected roots 

being less likely to bear root hairs (Chilvers and Daft, 1981). Iqbal and Firdaus 

(1986a,b) described mycorrhizae in foliage and dried sheathing leaves of bulbs of 

N. poeticus, as well as in the roots. 

The seedling and development of the Juvenile plant 

Germination and seedling growth were described by Chouard (1926, 1931) and 

Chan (1952). Germination is hypogeal. The cotyledonary sheath extends to 2–3 

cm before the primary, grass-like leaf breaks through and appears above ground. 

After the growth of the radical, one or two adventitious roots appear, and the radical 

is later lost. The base of the cotyledonary sheath swells to form the outermost scale, 

as does the base of the foliage leaf within it, itself enclosing the apical meristem. 

The contractile roots pull the bulb down. The one-year-old bulb is about 1 cm 

long. There is no information on environmental factors controlling bulbing in the 

seedling, although this takes place under increasing daylength (Rees, 1972). 

There is a juvenile phase of several years. In narcissus cultivars, the plant 

produces one or two scales and a single foliage leaf in its second and third years, 

a second leaf is produced in the fourth year, and three or more in the fifth (Chan, 

1952; Rees, 1972). Anthesis has been reported to occur after three to eight years, 

at the shorter end of this range for some species and after a longer time for largeflowered 

cultivars (Chan, 1952; Jamiolkowska and Zawadzka, 1971; Rees, 1972; 

Koopowitz, 1986). When a bulb fails to flower, growth is then similar to that of a 

younger bulb, and at least one foliage leaf is produced in addition to the usual 

complement, and all foliage leaves then have completely sheathing bases (Chan, 

1952). The growth and development of small bulbs propagated artificially (e.g., by 

chipping) is similar to that of seedlings of similar size. 

CONCLUSIONS 

Although ‘the processes which go on inside a bulb are very difficult to test’ 

(K. Goebel, quoted by Rees, 1972), the structure and development of the narcissus 

bulb have been well described, primarily by Chan (1952) and Rees (1969). On the 

other hand little is known of the endogenous or environmental factors which 

control processes such as bulb scale/foliage leaf differentiation or the initiation of 

bulb units or flowers. A few studies suggest there are varietal differences in floral 

initiation and bulb ‘scale’ numbers, while a few species have no cold requirement


or respond to ethylene. Further investigation of these features may facilitate the 

manipulation of narcissus growth and development, so that desirable characters – 

whether ornamental or industrial – could be exploited. 

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2 The folklore of Narcissus 

Anthony C. Dweck 

INTRODUCTION 

William Wordsworth probably did more for the daffodil than any other in his 

enchanting verse: 

I wandered lonely as a cloud 

That floats on high o’er vales and hills, 

When all at once I saw a crowd, 

A host of golden daffodils; 

Beside the lake, beneath the trees, 

Fluttering and dancing in the breeze. 

Continuous as the stars that shine 

And twinkle on the milky way, 

They stretched in never-ending line 

Along the margin of the bay: 

Ten thousand saw I at a glance, 

Tossing their heads in sprightly dance. 

John Gerard sets the scene ideally for this chapter. The fair Lady Europa, entering 

with her Nymphs into the meadows, did gather the sweet smelling daffodils: 

But when the Girles were come into 

The medowes flouring all in sight, 

That Wench with these, this Wench with those 

Trim floures, themselves did all delight: 

She with the Narcisse good in sent, 

And she with Hyacinths content. 

Gerard in his Herbal (Woodward, 1990) says ‘It is not greatly to our purpose, particularly 

to seek out their places of growing wild, seeing we have them all and 

every one of them in our London gardens, in great abundance. The common wild 

Daffodil groweth wild in fields and sides of woods in the West parts of England.’ 

Today, the daffodil or narcissus is a very popular garden plant and an important 

commercial crop, with a large number of species, hybrids and varieties in 

cultivation (Figure 2.1). Gerard’s Herbal lists 37 different types that were already 

in cultivation by the end of the l6th century, which demonstrates the popularity of

20 A.C. Dweck 

Figure 2.1 The narcissus, a plant with a rich folklore (from Bessette and Chapman, 

1992). 

the plant from the early days of horticulture. Many different daffodils are now 

found naturalised in grassland, hedge-banks, woodland margins, roadsides and 

waste ground throughout the British Isles, especially in the south. 

THE LANGUAGE OF FLOWERS 

In the Middle Ages, when the art of reading and writing was known only by a privileged 

few, there grew up a tradition of the language of flowers, whereby every 

flower had a meaning. It was a tradition that was revived by the early Victorians, 

who took great delight in this fanciful idea and collected together much of the 

information that survives to this day. The example that most of us would recognise

Folklore of Narcissus 21 

is the giving of red roses as a sign of love. In this tradition, the daffodil is for rebuttal 

in domestic situations: ‘I do not share your feelings’. However, in battle emblems 

the daffodil is for regard and chivalry (Greenaway and Marsh, 1978; Pickles, 1990). 

DERIVATION OF THE NAME DAFFODIL 

There are a number of thoughts on how the name daffodil came into being. The 

popular English names daffodowndilly, daffodily and affodily may be corruptions 

of asphodel, since the daffodil was thought to be identical with the blossoms 

mentioned by the ancient Greeks. Another school of thought is that the name 

comes from the Mediaeval Latin affodilus, Latin asphodilus or Greek asphodelus, which 

was the name of that plant which grew across the meadows of the underworld and 

belonged to Persephone, the Queen of Hell (Grigson, 1996). 

Pliny describes the narcissus as ‘narce narcissum dictum, non a fabuloso puero’, which 

translated means ‘named narcissus from narce, not from the fabulous boy’. The 

Greek narkao, meaning to be numb, originates in the narcotic properties of the 

plant (Genders, 1985). 

The popularity of the daffodil in the British Isles is attested by the large number 

of common names used in various parts of the country (Dony et al., 1986; Grigson, 

1996; Grieve, 1998). These include: 

Popular name Place 

Affodil, Affrodil Cheshire 

Bell-Flowers Dorset and Somerset 

Bell-Rose Somerset 

Butter and Eggs Devon, Somerset and Northampton 

Churn Lancashire 

Cowslip Devon 

Cuckoo-Rose Devon and Somerset 

Daffodil England, Scotland, Ireland 

Daffydowndilly Somerset 

Daffy-down-dilly Somerset 

Daffydilly Northamptonshire 

Dillydaffs Somerset 

Easter Lily Devon and Somerset 

Easter Rose Somerset 

Fairy Bells Dorset 

False Narcissus Devon 

Fleur de Coucou Devon 

Garden Narcissus Devon 

Giggary Devon 

Gylfinog Wales 

Gold Bells Wiltshire 

Golden Trumpets Somerset 

Gooseflop Somerset 

Goose-Leek Isle of Man 

Gracie Daisies Devon and Somerset 

Gracie Day Devon 

Hen and Chickens Devon 

Hoop Petticoats Dorset 

Jonquil Hertfordshire

22 A.C. Dweck 

Julians Hertfordshire 

King’s Spear Somerset 

Lady’s Ruffles Wiltshire 

Lent-Cocks Devon and Somerset 

Lent-Lily Cornwall, Devon, Dorset, Somerset, Isle 

of Wight, Gloucestershire, Sussex, Kent, 

Surrey, Suffolk, Warwickshire, Cheshire, 

Derbyshire, Lincolnshire, Yorkshire, 

Westmoreland 

Lent Pitchers Devon and Somerset 

Lent-Rosen Devon and Somerset 

Lents Cornwall, Devon, Lancashire 

Lenty Cups Somerset 

Lent Lily Cornwall 

Lily Scotland 

Narcissus Norfolk 

Porillon Norfolk 

Queen Anne’s Flowers Norfolk 

St Peter’s Bell Wales 

Sun-Sonnets Somerset 

Whit Sunday Devon 

Wild Daffodil Yorkshire 

Wild Jonquil Yorkshire 

Yellow Maidens Somerset 

Fleur d’asphodèle France 

Pauvres filles de Sainte Claire France 

MYTHOLOGY AND LEGEND 

According to Culpeper’s Herbal (Potterton, 1983), yellow daffodils are under the 

dominion of Mars. 

Daffodil flowers, though beautiful to the sight, leave a feeling of sadness when 

the history and folklore of the plant is examined. In classical mythology there was 

a handsome Greek shepherd boy named Narcissus. Though he was loved by all 

the wood nymphs, there was one called Echo who loved him more than the rest. 

Unfortunately, she could not tell him of her love, because she was only able to 

repeat his last words. It comes as no surprise to learn that Narcissus was totally 

unaware of Echo’s love and adoration for him. He was equally unaware of the pain 

and suffering that his ignorance of her love was causing her. Echo became thinner 

and thinner as her love robbed her of her appetite, until she slowly pined away to 

nothing more than a spirit who took sanctuary in the mountains. Only her soft voice 

remained. Venus, the goddess of love, came to hear of Echo’s hopeless devotion 

and immediately assigned the blame for her condition on Narcissus, who she 

decided should be punished. One day Narcissus was hunting in the forest. Little 

did he know that Venus had arranged with Cupid to set a magic spell on him so 

that he would fall in love with the first person that he saw. Coming to a crystal 

clear pool he stopped for a cooling drink to assuage his thirst and there in the 

water he saw another face rise up to meet his own as he leant over. Narcissus 

immediately succumbed to Cupid’s spell and fell in love. Again and again he tried 

to catch the face of the spirit who appeared to live in the water. In vain he called


out to this vision, but all that could be heard was the faint and sad echo coming 

from the mountains. Narcissus had fallen in love with his own reflection. Every 

day he returned to the pool in the hope of capturing the face that he saw there, 

and every day his tears added to the water in the pool. Slowly, like Echo, he began 

to waste away with unrequited love. The Immortals were not totally heartless and 

turned him into a delicate white papery flower, which would grow forever by the 

pool in memory of the egotistical youth. Another story continues by saying that 

when the nymphs came to look for him, they only found ‘A rising stalk with yellow 

blossoms crown’d’, and that the cup in the flower’s centre of all varieties contains 

the tears of Narcissus (Pickles, 1990). 

This story has led to the name being used as the term ‘narcissism’ or ‘narcissistic 

personality disorder’, in which people described by this condition have a grandiose 

view of their own uniqueness and abilities; they are preoccupied with fantasies of 

great success. To say they are self-centred is an understatement (Davison and Neale, 

1998). These characteristics have been validated in empirical studies (Ronnington 

and Gunderson, 1990) and are often a factor with borderline personality disorders 

(Morey, 1988). Such people are constantly seeking attention and adulation, and 

are, underneath, extremely sensitive to criticism and have a deep fear of failure. 

Many of the contemporary studies have been carried out by Heinz Kohut (Kohut, 

1971, 1977; Kohut and Wolf, 1978). 

The flower has another legend, which is even more gruesome than the former! 

Earth first put forth the flowers to lure the lovely Prosperine for Pluto, god of 

the underworld. The maid was so taken with the beauty of the daffodil that she 

stopped to admire it and as she stooped to pick it, the very worst happened. Pluto 

looking out from his hiding place took advantage of this momentary lack of attention 

and pounced out from his lair and seized her. It was, therefore, quite understandable 

why the ancients labelled the narcissus the flower of deceit. It was also the flower 

of imminent death, since it was the last bloom she plucked (MacFadyen, 1992). 

Another version of this story is told by Perdita in William Shakespeare’s The 

Winter’s Tale, where it was Proserpina who was picking lilies and was subsequently 

captured by Pluto. However, in this story, as she dropped the lilies in her fear, 

they turned into daffodils as they touched the ground. 

FOLKLORE AND RELIGIOUS CONNECTIONS 

Narcissus tazetta, which grows on the Plain of Sharon, Israel, may be the plant 

referred to in the biblical reference ‘ ...The wilderness and the solitary place shall 

be glad for them; and the desert shall rejoice, and blossom as the rose . . . ’ (Isaiah 

35 v.1). The Hebrew word here translated ‘rose’ may indicate a bulbous plant, 

rather than a rose (Tenney, 1967). 

Daffodils are considered by many to be unlucky, and they will not have the flowers 

in their house because they hang their heads, bringing tears and unhappiness. 

The sweet-scented old fashioned white narcissus, also called scented lily or white 

lily, is also known as grave flowers and unlucky to take indoors (Vickery, 1995). 

In the Isle of Man it is unlucky to have the plant in the house till the goslings have 

hatched. The Manx name is lus-ny-guiy or goose herb. In common with primroses, 

daffodils were sometimes banned from the house by poultry-keepers, and, in

24 A.C. Dweck 

Herefordshire, if daffodils are brought in when the hens are sitting, they say there 

will be no chickens. However in Devon, the number of goslings hatched and 

reared is said to be governed by the number of wild daffodils in the first bunch of 

the season brought into the house (Vickery, 1995). 

Robert Herrick alludes in his Hesperides to the daffodil as a portent of death, 

probably connecting the flower with the asphodel, which the ancient Greeks 

planted near tombs. Despite this he writes ‘Fair daffodils, we weep to see/You 

haste away too soon . . . ’ 

The occurrence of wild daffodils is sometimes said to indicate the former site of 

a religious foundation. At Frittlestoke, near Torrington, Devon, it was recorded in 

1797 that the people of the village call daffodils by the name Gregories, a name 

that coincided with the order of a neighbouring monastery – the Canons of St 

Gregory (Britten and Holland, 1886). In both Hampshire and the Isle of Wight it 

was generally said that wild daffodils indicated the site of a monastery. St Urian’s 

Copse is well known for its primroses and daffodils. There is a tradition that daffodils 

grow in profusion on one side of a track running through the copse because a 

religious building once stood there. The only sizeable population of wild daffodils 

in the London area is found at Abbey Wood, named after Lesney Abbey (Vickery, 

1995). 

HISTORICAL TALES 

A crusader returned home to Churchill (in Avon, in the west country of England), 

having spent years fighting the Crusades in the Holy Land. A rich man before his 

departure, he had returned home poor. His wife was a lover of precious and rare 

flowers, and so he had carefully brought back with him two bulbs of the Primrose 

Peerless. The story is a sad one, since when he returned, it was to a wife who had 

been buried for four years. In despair he flung the cherished bulbs over the 

churchyard wall. He is said to have died of a broken heart. However, throughout 

the centuries the bulbs have grown and flourished and kept his memory alive 

(Vickery, 1995). 

Both the daffodil and the leek are national symbols of Wales. The daffodil is 

associated with St David because it is traditionally said to bloom first on his day 

(1 March). It is an easier emblem to wear than the leek, and many a schoolchild in 

Wales sports one, real or artificial, on this date (Vickery, 1995). 

Since 1990, National Daffodil Day has been promoted by Marie Curie Cancer Care. 

At about the same time, the Irish Cancer Society similarly adopted the daffodil as a 

symbol. In Australia, they also have a national fund raising day for cancer research 

(Anon., 1997). 

On the Isles of Scilly, The Prince of Wales is paid one daffodil annually as rent 

for the untenanted lands of Scilly, paid by the local Environmental Trust. 

HERBAL MEDICINES 

Considering that narcissus are a rich source of alkaloids (see Chapter 6, this 

volume), it is not surprising that the genus has figured in herbal medicine. This


has been vindicated by recent developments. The Daily Mail (28 September 1996) 

carried a headline ‘Shire says it with snowdrops’. ‘Flower power could soon be 

helping sufferers of chronic fatigue syndrome. Shire Pharmaceuticals is testing 

galanthamine, a compound found in daffodils and snowdrops, on victims of “yuppie 

flu”. The drug already has improved the mental performance of Alzheimer’s 

patients.’ 

The Greek physician Hippocrates of Cos (460–377 BC) recommended a pessary 

prepared from Narcissus oil (probably N. poeticus) for the management of uterine 

tumours (Pettit et al., 1986). Plants of the Narcissus genus have been used to treat 

a variety of human medical problems (Pettit et al., 1995), and N. poeticus was 

described in the Bible as a well-established treatment for symptoms that would 

now be defined as cancer (Pettit et al., 1990). Pliny the Elder (AD 23–77) also 

recorded the topical use of N. poeticus and another derived from N. pseudonarcissus 

for the treatment of uterine tumours. It is now known that N. poeticus contains 

0.012% of the antineoplastic agent narciclasine in the fresh bulb (Piozzi et al., 

1969). 

However, narcissus are not recommended for domestic use. A homoeopathic 

medicine is made from the bulbs and used for respiratory disease, particularly 

bronchitis and whooping cough, according to Culpeper’s (1616–1654) Herbal 

(Potterton, 1983): 

The roots boiled and taken in posset drink cause vomiting and are used with 

good success at the appearance of approaching agues, especially the tertian 

ague, which is frequently caught in the springtime. A plaster made of the roots 

with parched barley meal dissolves hard swellings and imposthumes, being 

applied thereto; the juice mingled with honey, frankincense wine, and myrrh, 

and dropped into the ears is good against the corrupt and running matter of 

the ears, the roots made hollow and boiled in oil help raw ribed heels; the 

juice of the root is good for the morphew and the discolouring of the skin. 

Galen [AD 130–201] saith: That the roots of Narcissus have such wonderfull 

qualities in drying, that they consound and glew together very great wounds, 

yea and such gashes or cuts as happen about the veins, sinues, and tendons. 

They have also a certaine clensing facultie. The root of Narcissus stamped with 

hony and applied plaisterwise, helpeth them that are burned with fire, and 

joineth together sinues that are cut in sunder. Being used in manner aforesaid 

it helpeth the great wrenches of the ancles, the aches and pains of the joints. 

The same applied with hony and nettle seed helpeth Sun burning. Being 

stamped with the meale of Darnel and hony, it draweth forth thorns and stubs 

out of any part of the body. 

Narcissus are also referred to in John K’Eogh’s Irish Herbal (Scott, 1986). Narcissus 

was said to have a hot and dry nature. The roots, pounded with honey were good 

against burns, bruised sinews, dislocations and old aches. They take away freckles 

and heal abscesses and sores, and they draw out thorns and splinters. A decoction 

of the roots is a great emetic. 

It has also been used as an application to wounds, for hard imposthumes, for 

strained sinews, stiff or painful joints, and other local ailments. The narcissus was

26 A.C. Dweck 

the basis of an ancient ointment called Narcissimum. The powdered flowers have 

been used as an emetic in place of the bulbs, and in the form of a syrup or infusions 

for pulmonary catarrh. A decoction of the dried flowers acts as an emetic, and has 

been considered useful for relieving the congestive bronchial catarrh of children, 

and also useful for epidemic dysentry. In France, narcissus flowers have been used 

as an antispasmodic. A spirit has been distilled from the bulb, used as an embrocation 

and also given as a medicine and a yellow volatile oil, of disagreeable odour and 

a brown colouring matter has been extracted from the flowers, the pigment being 

quercetin, also present in the outer scales of the onion. The Arabians commended 

the oil to be applied for curing baldness and as an aphrodisiac (Grieve, 1998). 

Conveniently, the bulbs of N. tazetta have also been used as a contraceptive (Matsui 

et al., 1967). The influence of daffodil on the nervous system has led to giving its 

flowers and bulb for hysterical affections and even epilepsy, with benefit. It 

entered into the books as a purge and a vomitive and a cure for erysipelas and the 

palsy (Grigson, 1996). 

Throughout the Middle Ages, the Arabian, North African, Central American and 

Chinese medical practitioners continued to use Narcissus oil in cancer treatment 

(Pettit et al., 1993). For example, the bulbs of N. tazetta var. chinensis, cultivated in 

China as a decorative plant, were also used topically in folk medicine as a liniment 

for the treatment of tumours. In this case, pretazettine was proved to be one of the 

antitumour active compounds (Furusawa et al., 1973; Ma et al., 1986). The bulbs of 

N. tazetta continued to be used in Turkey as a home remedy for the treatment of 

abscesses, because of their antiphlogistic and analgesic property (Çakici et al., 1997). 

POISONOUS EFFECTS 

Socrates called the narcissus the ‘Chaplet of the infernal Gods’, because of its 

narcotic effects. An extract of the bulbs, when applied to open wounds, has 

produced staggering, numbness of the whole nervous system and paralysis of the 

heart (Grieve, 1998) 

There have been cases of poisoning when the bulbs have been eaten in mistake 

for onions (Culpeper’s Herbal; Potterton, 1983). Lycorine or narcissine in warmblooded 

animals acts as an emetic, causing eventual collapse and death by paralysis 

of the central nervous system: cattle, goats and pigs have been poisoned by the 

plant (Manning, 1965). With cats, narcissine causes nausea and purgation (Grieve, 

1998). The poison acted speedily, high temperature did not destroy the toxicity of 

the poison and only a relatively small amount was needed (Grieve, 1998). Ingestion 

of narcissus bulbs produces severe gastroenteritis and nervous symptoms, apparently 

owing to the phenanthridine alkaloids contained therein (Tyler et al., 1988). 

When the bulbs have been mistaken for onions and eaten, either raw or cooked, 

symptoms including dizziness, stomach pains, nausea, vomiting and diarrhoea 

have developed shortly afterwards. In more severe poisoning there may be trembling, 

convulsions and paralysis. Vomiting has occurred in children who have 

eaten a few leaves, and there is also a report of a four-year-old child who died after 

sucking a narcissus stalk. Recovery, however, is usually complete in a few hours 

without any treatment being necessary. Those who pick and pack the flowers are 

liable to develop dermatitis, probably caused partly by the irritant effects of the sap


and partly by an allergic reaction. Animals rarely eat these plants, although, during 

the food shortage in the Netherlands in the Second World War, some cattle died 

after being given narcissus bulbs to eat. A tortoise which ate four daffodil leaves 

lost its appetite and became constipated and listless; it died 11 days later. In severe 

cases it may be necessary to induce vomiting or remove stomach contents (Cooper 

and Johnson, 1991). In South Africa, similar problems with toxicity are experienced. 

The bulbs of daffodil and narcissus are known to have caused death when 

eaten by mistake (Moll and Moll, 1989). A case of poisoning by daffodil bulbs, 

cooked by mistake in the place of leeks, was reported from Toulouse in 1923. The 

symptoms were acute abdominal pains and nausea, which yielded to an emetic 

(Grieve, 1998). The bulbs of Narcissus poeticus, the Poet’s narcissus, are reported to 

be more dangerous than those of the garden daffodil, being powerfully emetic and 

irritant (Grieve, 1998). 

ENDANGERED SPECIES 

Large garden varieties of daffodils have recently been crossed with many of the 

small wild species to produce delightfully graceful blossoms. Narcissus triandrus and 

N. cyclamineus have been used in breeding for many years. The first species makes 

small clusters of blooms with very silky petals. N. cyclamineus usually gives genes for 

flowers with backswept petals, a long-waisted trumpet, and early flowering. These 

types of daffodils are constantly popular and the demand for miniature daffodils 

far exceeds the supply. Unfortunately, the ability to produce new kinds of miniature 

daffodils is hampered by the disappearance of many of the tiny wild species. 

N. calcicola, a tiny yellow jonquil from Spain and Portugal is considered endangered 

and N. watieri, possibly the most powerful tool for making miniature white daffodils, 

is already unobtainable (Koopowitz and Kaye, 1990). Occasionally N. watieri from 

North Africa’s Atlas Mountains is advertised by unscrupulous bulb merchants who 

substitute another variety. The exact status of N. watieri is unknown. The only 

hope is that a few plants may still exist on some of the rocky ledges or hillsides 

where the species once thrived. If so, perhaps it can be introduced once again. More 

likely, the species has already been destroyed – either over-collected or eaten by 

goats (Koopowitz and Kaye, 1990). 

FOOD USE 

On the upper Nile, Grant found a narcissus about 20 cm high, with white flowers 

having a waxy, yellow corona and with leaves tasting of onions. The leaves, cooked 

with mashed groundnuts, he reported, make a delicious spinach (Hedrick, 1972). 

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Designers. Dover Publications, New York.

28 A.C. Dweck 

Britten, J. and Holland, R. (1886) A Dictionary of English Plant Names. Publisher unknown, 

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Inc., New York, pp. 340–341. 

Dony, J.G., Jury, S.L. and Perring, F.H. (1986) English Names of Wild Flowers. 2nd edition. 

The Botanical Society of the British Isles, Reading. 

Furusawa, E., Suzuki, N., Tani, S., Furusawa, S., Ishioka, G.Y. and Motobu, J. (1973) 

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Biology and Medicine, 143, 33–38. 

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Bower, Exeter. 

Greenaway, K. and Marsh, J. (1978) The Language of Flowers. Holt, Rinehart and Winsher. 

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Ma, G.E., Li, H.Y., Lu, C.E., Yang, X.M. and Hong, S.H. (1986) 6α/6β-Hydroxy-3- 

O-methylepimaritidine, two new alkaloids from Narcissus tazetta L. var. chinensis Roem. 

Heterocycles, 24, 2089–2092. 

Matsui, A.D.S., Rogers, J., Woo, Y.K. and Cutting, W.C. (1967) Effects of some natural 

products on fertility in mice. Medicina et Pharmacologia Experimentalis, 16, 414. 

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agents, 162. Zephyranthes candida. Journal of Natural Products, 53, 176–178. 

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genus Hymenocallis. Journal of Natural Products, 58, 756–759. 

Pickles, S. (1990) The Language of Flowers. Publisher not given, London.


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metabolites in Amaryllidaceae. Phytochemistry, 8, 1745–1748. 

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Febiger, Philadelphia. 

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Woodward, M. (editor) (1990) Gerard’s Herbal. Studio Editions, London.

3 Classification of the genus Narcissus 

Brian Mathew 

CLASSIFICATION OF NARCISSUS SPECIES 

It is generally acknowledged that the genus Narcissus presents great taxonomic 

problems, and there have been numerous attempts at its classification. Some 

authors have taken a very wide view of the concept of each species (e.g., Webb, 

1980), resulting in as few as 26 recognised species, while some (e.g., Fernandes, 

1969b) have taken a very narrow view which results in the recognition of a great 

many species (upwards of 60), often involving a complex hierarchy of infraspecific 

taxa. At the generic level, some researchers have taken a narrow view of the 

delimitation of genera; for example, in the system devised by Haworth (1831), 

many of the presently accepted subgenera, sections and subsections were recognised 

as separate genera (e.g., Hermione for the ‘tazetta’ group, Corbularia for the 

‘bulbocodium’ group, Ajax for the ‘pseudonarcissus’ group, Ganymedes for the 

‘triandrus’ group, etc.). On the other hand, Herbert (1837) took a rather wider 

view of the genera, and reduced the 16 genera recognised by Haworth to six, 

while Spach (1846) went a stage further and treated many of Haworth’s genera 

as sections of the one genus, Narcissus. Many of the plants that had been 

described as species up to that time were of unknown (or garden) origin, and it 

was Baker (1875) who attempted to clarify the situation by considering only wild 

source material for his classification. All the segregate genera recognised by most 

of the previous authors were included by him in Narcissus, except for Tapeinanthus, 

which Baker regarded as sufficiently distinct to uphold at generic level; 

nowadays it, too, is often ‘sunk’ into Narcissus (e.g., Webb, 1980), although Cullen 

(1986) maintains it on the basis of the rudimentary corona and near-absence of a 

perianth tube. 

Although the status (subgenus, section, etc.) bestowed upon the individual groups 

may vary somewhat from author to author, as does the status of the individual 

taxa within the groups, the actual content of each group is similar in the various 

classifications. Most of the groups – most frequently referred to as sections – are 

fairly obvious, for example the Trumpet daffodils, the Tazettas, the Pheasant’s 

Eyes, the Hoop Petticoats, the Jonquils, and so on, and these are the basic divisions 

in the genus recognised here. It is, however, considered somewhat unsatisfactory 

to give all the infrageneric groupings similar status (in some recent classifications, 

e.g., Webb (1980), they are treated equally as sections). The classification used here 

is a combination and adaptation of the systems devised by previous researchers, 

notably those of Fernandes (1951, 1969a,b) and Webb (1980), in an attempt to

Classification of Narcissus 31 

reflect the relationships more clearly. Some examples of species representing the 

main sections are illustrated in Figure 3.1. 

An additional complication to the taxonomy is posed by hybridisation. Most 

species of Narcissus will hybridise but, significantly, there is great variation in the 

fertility of the offspring, depending upon the degree of relationship between 

the parents. The important cytological work by Fernandes (1951, 1969a,b), 

Brandham and Kirton (1987) and Brandham (1992) has done much to clarify the 

genetics of the genus. There has been a great deal of hybridisation in this very 

popular, garden-worthy genus, resulting in thousands of hybrid cultivars and 

selections (Kington, 1998), and doubtless this will continue. Although much of this 

work has been concerned with sophisticated selection for flower form and colour 

(e.g., pink and red coronas and apricot-coloured perianth segments), there are 

probably still some interesting lines of research that could be pursued using the 

many wild species. Taking just one possibility as an example, the autumn-flowering 

species (Narcissus serotinus, N. elegans and the green-flowered N. viridiflorus) could 

perhaps be utilised in the production of a race of larger-flowered autumnal 

narcissi, thus extending the overall flowering season of the garden forms by 

several months. With the great diversity of characters exhibited by the species and 

their numerous variants, there are great possibilities in this natural gene pool. 

However, some of the species are under threat in the wild, and many more will 

become so with increasing urban and tourist-based development. A good example 

is provided by N. cyclamineus, which in recent decades has been one of the most 

important species in the production of a wide range of ‘Cyclamineus’ daffodils of 

Division 6 in the Horticultural Classification: this species may already be extinct in 

the wild, or at best very scarce (Blanchard, 1990). Even in the case of widespread 

and well-known species, there are often local variants which could be of potential 

in breeding programmes. It is thus essential that steps are taken to ensure the 

survival of these wild progenitors of the garden daffodils, so that this valuable 

gene pool is not lost or severely depleted. 

Although the family Amaryllidaceae as a whole may be seen as primarily tropical 

or subtropical in its distribution, the genus Narcissus is to be found largely in 

south-western Europe, notably Spain and Portugal, and in North Africa. A few 

species extend into France and Italy, and even fewer are found farther east in the 

Balkans (N. poeticus, N. serotinus and N. tazetta) and the eastern Mediterranean 

(N. serotinus). Records outside this area – for example, N. tazetta variants in western 

and central Asia, China and Japan – are almost certainly ancient introductions. 

The extent of the natural distribution northwards is also unknown: although there 

are apparently wild populations of N. pseudonarcissus or similar species in Britain, 

for example, these could well be the result of early introductions that have become 

naturalised. 

OUTLINE OF CLASSIFICATION 

1. Subgenus Narcissus 

a. Section Narcissus 

b. Section Pseudonarcissus 

c. Section Ganymedes

(1) 

(2) 

(3)

(4) 

(5) (6) 

Figure 3.1 Examples of Narcissus species of the main sections: (1) Subgenus Narcissus, 

Section Narcissus: N. radiiflorus; (2) Subgenus Narcissus, Section Pseudonarcissus: 

N. cyclamineus; (3) Subgenus Hermione, Section Hermione: N. papyraceus; (4) Subgenus 

Corbularia: N. romieuxii; (5) Subgenus Narcissus, Section Ganymedes: N. triandrus; 

(6) Subgenus Narcissus, Section Jonquillae: N. gaditanus. Photos: Brian Mathew. 

(See Colour plate 1)

34 B. Mathew 

d. Section Jonquillae 

i. Subsection Jonquillae 

ii. Subsection Apodanthi 

iii. Subsection Chloranthi 

e. Section Tapeinanthus 

2. Subgenus Hermione 

a. Section Hermione 

i. Subsection Hermione 

A. Series Hermione 

B. Series Albiflorae 

ii. Subsection Angustifolii 

iii. Subsection Serotini 

b. Section Aurelia 

3. Subgenus Corbularia 

CONSPECTUS OF THE GENUS NARCISSUS 

1. Subgenus Narcissus [Subgenus Eu-narcissus Pax] 

Description: Flowers usually vernal, rarely autumnal, umbels solitary- to severalflowered. 

Perianth segments well-developed; corona well-developed, trumpet-shaped, 

cup-shaped or rudimentary; perianth tube varying from ± absent to funnel-shaped 

or long and slender; stamens with ± straight filaments. Chromosome number x = 7 

(usually 2n = 14). 

Five sections of subgenus Narcissus are recognised, Narcissus, Pseudonarcissus, 

Ganymedes, Jonquillae and Tapeinanthus. 

1a. Section Narcissus [Section Helena (Haworth) Ascherson and Graebner] 

Description: Leaves linear or lorate, usually grey-green, flat or channelled. Flowers 

vernal, solitary, fragrant; perianth tube cylindrical; segments spreading or slightly 

reflexed; corona shallow, disk-like or very widely funnel-form, much wider than 

deep. Filaments much shorter than the anthers; anthers dorsifixed, the lower 

three usually included and the upper three at least partly exserted. 

Type species: N. poeticus L. 

Note: There have been great differences of opinion over the classification of the 

Poet’s or Pheasant’s Eye narcissus. At one extreme all the variations are considered 

to belong to one species, N. poeticus, while at the other extreme about ten separate 

species have been recognised. The current view, which appears to have gained 

most support, is that there are two species, N. poeticus and N. radiiflorus, with several 

named variants of each. 

N. poeticus L. Petals broad and overlapping at the base, giving a substantial 

‘rounded’ flower; stamens unequal. 

var. poeticus. Corona flattish and disk-like; petals at right angles to corona; 

flowers up to 7 cm diameter. Southern France, Italy.


var. hellenicus (Pugsley) Fernandes. Corona shallow cup-shaped; petals 

reflexed; flowers small, about 4.5 cm diameter. Greece. 

var. majalis (Curtis) Fernades. Corona shallow cup-shaped, white-zoned below 

the red edge; flowers up to 7 cm diameter. France. 

var. recurvus (Haworth) Fernandes. Corona shallow cup-shaped, green-yellow 

edged red; petals reflexed; flowers up to 7 cm diameter. Switzerland. 

var. verbanensis Herbert. Corona shallow cup-shaped; petals very pointed at 

apex; flowers small, about 3.5–5 cm diameter. Italy. 

N. radiiflorus Salisbury. Petals not overlapping at the base, the flower more 

‘starry’ in appearance; stamens nearly equal. 

var. radiiflorus. Corona shallow cup-shaped, less than 1 cm across. Switzerland, 

Austria, northern Balkan Peninsula. 

var. stellaris (Haworth) Fernandes. Corona shallow cup-shaped and about 

1 cm across. Central and eastern Europe. 

var. exertus (Haworth) Fernandes. Corona flattish and disk-like, yellow or 

yellowish-green with a red or orange edge. Switzerland, southern France. 

var. poetarum Burbidge and Baker. Corona flattish, wholly red. Of unrecorded 

origin. 

1b. Section Pseudonarcissus DC. [subgenus Ajax Spach] 

Description: Leaves usually grey-green, flat. Flowers vernal, usually solitary, rarely 

up to four, wholly yellow, wholly white or bicoloured; perianth tube broadly conical; 

segments usually spreading, rarely suberect or much reflexed; corona cylindrical, 

often widened at the mouth, much longer than wide. Filaments straight, subequal 

to or much longer than the anthers; anthers ± basifixed or rarely dorsifixed, 

exserted from the perianth tube, included within the corona. 

Type species: N. pseudonarcissus L. 

Note: These are the true daffodils in which the corona is in the form of a long more 

or less cylindrical trumpet rather than a cup, saucer or funnel. They tend to flower 

in mid spring, after the early Hoop Petticoats and Angel’s Tears but before the 

N. poeticus forms. Numerous taxa have been described in this section and these 

have been accorded various taxonomic ranks, many of them as variants of 

N. pseudonarcissus. An added complication to the taxonomy is that they have been 

cultivated for centuries and many plants of unrecorded origin have also been 

described as species. A very thorough revision of the group is required. They are 

listed below in two groups, the very small ones such as N. asturiensis and N. nanus 

and the larger-flowered including N. pseudonarcissus and its relatives. 

Group A. Plants small, usually less than 15 cm; flowers to 3.5 cm diameter 

N. asturiensis ( Jordan) Pugsley. Plant to 10 cm in height. Leaves grey-green. Flowers 

yellow; corona constricted in the middle. Northern Portugal, north-central and 

north-west Spain, on acidic soils. Var. villarvildensis Diaz and Prieto is a slight variant 

from central-northern Spain, as is var. brevicoronatus Pugsley which has a shorter 

corona.

36 B. Mathew 

N. cyclamineus DC. Flowers yellow, with sharply reflexed petals. Leaves bright 

green, not at all glaucous. North-west Portugal, north-west Spain. 

N. jacetanus Casas. 

var. jacetanus. Very similar to N. asturiensis but with a very incised corona. 

Northern Spain, on limestone formations. 

var. vasconicus (Casas) Casas. A very small-flowered variant, corona only 

1.5 cm long. Northern Spain, on acid formations. 

N. lagoi Merino. Up to 50 cm tall but with small flowers, similar to those of N. asturiensis. 

North-west Spain. 

N. minor L. Similar to N. asturiensis but a larger plant to 15 cm with broader leaves. 

Flowers yellow, the corona not constricted in the middle. Long cultivated, and of 

unknown origin, possibly the result of hybridisation between N. asturiensis and 

N. pseudonarcissus. 

N. nanus Spach. Early-flowering, bicoloured with pale yellow petals and a deeper 

yellow, straight-sided corona. A small trumpet daffodil of unknown origin. 

N. provincialis Pugsley. Similar to N. minor, the flowers slightly larger to 4 cm long. 

Southern France. 

N. parviflorus Jordan. Flowers ± pendent, with pale cream-yellow perianth segments 

and deep yellow corona. South-western France. 

N. pumilus Salisbury. Similar to N. nanus but flowers wholly bright yellow, the 

corona very expanded and frilled at the margin. Origin unknown. 

N. portensis Pugsley. Up to 20 cm in height with uniform bright yellow flowers; 

perianth segments noticeably shorter than the funnel-shaped trumpet. Northern 

Portugal, central and north-western Spain. 

Group B. Plants often 15–60 cm or more and/or with large flowers, usually 

5–12 cm diameter 

N. abscissus (Haworth) Schultes and Schultes fil. Flowers bicoloured with cream 

perianth segments and yellow, parallel-sided corona not expanded at mouth. 

North-eastern Spain, south-western France. 

N. albescens Pugsley. Flowers creamy-white with a pale creamy-yellow corona, 

held horizontally; corona much-expanded at the mouth. Origin unknown. 

N. alpestris Pugsley. Flowers wholly white, pendent; perianth segments held 

alongside corona. North-eastern Spain. 

N. bicolor L. Similar to N. abscissus but with pale yellow perianth segments and 

deeper yellow corona. Origin unknown. 

N. bujei (Casas) Casas. Similar to N. hispanicus (below) but leaves less noticeably 

blue-green. Flowers wholly yellow, to 6.5 cm across; corona mouth not markedly 

frilled. Southern Spain. 

N. calcicarpetanus Casas. Flowers suberect or horizontal, wholly yellow; corona 

frilled 2.5–3 cm long, longer than perianth segments. North-central Spain. 

N. confusus Pugsley. Similar to N. hispanicus (see below) but with green leaves and 

corona less flared at mouth. Flowers wholly yellow, facing obliquely upwards. 

Central Spain. 

N. fontqueri Casas and Rivas Ponce. Related and very similar to N. nobilis (below). 

Northern Spain.


N. gayi (Hénon) Pugsley. Probably a variation of N. pseudonarcissus (below). Origin 

unknown. 

N. genesii-lopezii Casas. Described as a very glaucous-leafed plant with sulphurcoloured 

flowers. East-central Spain. 

N. hispanicus Gouan. Tall, with flowers of 10 cm diameter, yellow; leaves bluish, 

twisted; corona very flared at mouth. Long-cultivated, origin obscure. 

N. longispathus Pugsley. Tall with up to three usually wholly yellow flowers per 

umbel; spathes very conspicuous, to 10 cm long. South-eastern Spain. 

N. macrolobus (Jordan) Pugsley. Up to 25 cm tall; pale yellow with corona slightly 

deeper yellow; probably a variation of N. pallidiflorus. North-eastern Spain, southwestern 

France. 

N. moleroi Casas. Similar to N. alpestris but flowers smaller, pale yellow. North-eastern 

Spain. 

N. moschatus L. Up to 30 cm tall; flowers creamy-white, pendent with petals 

drooping alongside corona. Origin unknown, but probably from south-western 

France or north-eastern Spain. 

N. nevadensis Pugsley. Flowers up to four in an umbel, bicoloured with pale yellow 

petals and corona slightly deeper. Southern Spain. 

N. nobilis (Haworth) Schultes fil. Only 15–30 cm tall but flowers 8–12 cm diameter, 

bicoloured; petals white, corona deep yellow. Northern Spain, northern 

Portugal. Var. leonensis (Pugsley) Fernandes from north-central Spain is a largerflowered 

variant, with flowers up to 12.5 cm diameter with a very widely-flared 

corona. 

N. obvallaris Salisbury. To 30 cm but with relatively small flowers (4 cm 

diameter), wholly yellow. Central-southern Spain, naturalised in western 

Britain. 

N. pallidiflorus Pugsley. Flowers very pale cream/yellow, to 7.5 cm diameter; perianth 

segments twisted, corona mouth much-toothed. Northern and north-eastern 

Spain, south-western France. 

N. perez-chiscanoi Casas. Probably rather similar to N. obvallaris (above), with fairly 

small flowers. Central-south Spain. 

N. primigenius (Suarez ex Lainz) Casas and Lainz. This appears to be very similar 

to N. nobilis, perhaps best treated as a variant of it. North-west Spain. 

N. pseudonarcissus L. 

ssp. pseudonarcissus To 30 cm; bicoloured white/yellow flowers; corona not 

widely expanded and frilled. Spain, southern France, northern Italy; naturalised 

in several other countries in Europe. 

ssp. eugeniae (Casas) Casas. A smaller plant to 10 cm but flowers to 7.5 cm 

diameter; perianth segments not overlapping; leaves short and wide. Centraleastern 

Spain. 

N. pseudonarcissus ssp. pugsleyanus Barra and Lopez. Appears to be closer to 

N. nobilis, but with smaller flowers and a perianth tube that is markedly shorter 

(only 1.2–2 cm). Central Spain. 

N. radinganorum Casas. Probably close to N. hispanicus with wholly yellow flowers, 

but a smaller plant, 25–40 cm tall. South-eastern Spain. 

N. tortuosus Haworth. Flowers sulphur-white with twisted petals, similar to 

N. moschatus. Northern Spain.

38 B. Mathew 

1c. Section Ganymedes Salisbury ex Schultes and Schultes fil. 

Description: Leaves narrow, flat or ± cylindrical, usually dark green and slightly 

glaucous. Flowers vernal, one to about six in an umbel; perianth tube funnel-form; 

segments sharply reflexed; corona campanulate or obconical, about as wide as 

deep. Filaments of differing lengths, the lower three shorter than the anthers and 

with the anthers included, the upper three much longer than the anthers and the 

anthers exserted. 

Type species: N. triandrus L. 

Note: A very distinctive group, known as the Angel’s Tears daffodils. They 

have pendent flowers with sharply reflexed perianth segments; it is the three 

prominent upper stamens that have given rise to the epithet triandrus. The 

taxa in this group have been treated as separate species by some authors, or at 

various infraspecific levels within N. triandrus by others. The latter view is 

accepted here, but it must be pointed out that even these taxa are not well 

defined. 

N. triandrus L. 

var. triandrus. Flowers white or with a yellowish tinge on petals; leaves 

4–5 mm wide. Northern Spain. 

var. cernuus (Salisbury) Baker. Flowers creamy-white or very pale yellow; 

leaves very narrow, about 2 mm wide. Widespread in Spain and Portugal. 

var. concolor (Haworth) Baker [syn. N. triandrus ssp. pallidulus (Graells) 

Webb]. Flowers bright yellow. Portugal, central, southern and eastern 

Spain. 

var. loiseleurii (Rouy) Fernandes [syn. N. triandrus ssp. capax (Salisb.) Webb]. 

Similar to var. triandrus with white flowers; grows in sand by the sea on the 

Isles de Glenán, western France. 

Notes: The most frequently cultivated variant of this popular Narcissus has flowers 

of a cream colour and is known as ‘N. triandrus albus’; it is of unknown origin and 

should probably be regarded as belonging to var. cernuus. 

N. lusitanicus Casas Dorda and Casas from central Portugal may belong here 

with N. triandrus, since it is described as having affinities with N. pallidulus, which is 

regarded by Webb (1980) as a synonym of var. concolor; it is said to have intensely 

yellow flowers. 

1d. Section Jonquillae De Candolle 

Description: Leaves narrow, dark green, cylindrical or subcylindrical. Flowers 

vernal or rarely autumnal, one to several in an umbel, very fragrant; perianth 

tube cylindrical, slightly widened towards the apex; segments spreading or 

slightly reflexed; corona a shallow cup, wider than deep. Filaments shorter 

than anthers; anthers dorsifixed, included within the corona or slightly 

exserted. 

Type species: N. jonquilla L. 

Three subsections of section Jonquillae are recognised, Jonquillae, Apodanthi and 

Chloranthi.


1d(i). Subsection Jonquillae 

Description: Spring-flowering; flowers yellow throughout; corona cup-shaped, to 

6 mm deep. 

Type species: N. jonquilla L. 

N. jonquilla L. Leaves dark green, ± terete, channelled on the adaxial surface, 

1–4 mm wide. Flowers up to five in each umbel, fragrant, wholly yellow, 3–3.5 cm 

diameter; perianth tube straight, 2–3 cm long; segments spreading or slightly 

reflexed; corona 2–4 mm deep, 10–15 mm wide, shallowly crenate at the margin. 

Spain, Portugal. Note: The Jonquils are a confusing group with much natural 

variation. Botanists have for long disagreed as to how many species should be 

recognised. Listed below are the various taxa that have been described. 

N. assoanus Dufour [syn. N. juncifolius auct. plur., N. requienii Roemer]. Flowers 

one or two per umbel, 1.5–2.2 cm diameter; corona 3–5 mm deep, 9–11 mm wide; 

tube straight. North-eastern and eastern Spain, southern France. Ssp. praelongus 

Barra and Lopez has a longer perianth tube (to 27 mm). Southern Spain. 

N. baeticus Casas. Similar to N. assoanus but with a longer, narrower perianth tube. 

Southern Spain. 

N. cerrolazae Ureña. Flowers one to four, 2.5–4 cm diameter; corona 5–7mm 

deep, 11–16 mm wide; tube straight or curved. Southern Spain. 

N. cordubensis Casas. Flowers up to three, about 3 cm diameter; corona 5 mm 

deep, 15 mm wide; tube slightly curved. Southern Spain. 

N. fernandesii G. Pedro. Flowers up to four per umbel, 2.5–3.3 cm diameter; 

corona 6 mm deep, 8 mm wide; tube curved. South-central Portugal, south-central 

Spain. Var. rivas-martinezii (Casas) Casas from southern Spain is a variant with a 

straight perianth tube. 

N. gaditanus Boiss. and Reuter. Flowers up to eight per umbel, 1–2 cm diameter; 

corona 2.5–7 mm deep, 4.5–8 mm wide; tube curved. Southern Spain, southern 

Portugal. 

N. jonquilla var. henriquesii Sampaio. Flowers one or two per umbel, 3.5–4cm 

diameter; corona 6 mm deep, 10 mm wide; tube straight. South-central Portugal. 

N. marianicus Casas. Similar to N. fernandesii and probably best regarded as a 

variant of it. Central Spain. 

N. minutiflorus Willkomm. Probably should be regarded as a small variant of 

N. gaditanus with flowers only 1 cm across. Southern Portugal, southern Spain. 

N. palearensis Romo. This is described as being like N. jonquilla but usually with 

one to three flowers with corona 6–7 mm deep. North-eastern Spain 

N. pallens Freyn and Willk. This is now regarded by Fernandez Casas as a pale 

version of N. assoanus, var. pallens. 

N. willkommii (Sampaio) Fernandes. Flowers up to three per umbel, 1.5–2cm 

diameter; corona 4–5 mm deep, 7–11 mm wide; tube straight. Southern Portugal, 

south-western Spain. 

1d(ii). Subsection Apodanthi (A. Fernandes) D.A. Webb 

Description: Leaves usually grey-green, with two or four keels on the abaxial 

surface, channelled on the adaxial surface. Flowers vernal, one to several in an

40 B. Mathew 

umbel, fragrant; perianth tube cylindrical or slightly widened towards the apex; 

segments spreading or slightly reflexed; corona cup-shaped, wider than deep. 

Filaments shorter than the anthers; anthers dorsifixed, included within the corona 

or the upper whorl of three exserted. 

Type species: N. rupicola Dufour. 

Note: Although very similar to the species of subsection Jonquillae in flower shape, 

having long-tubed fragrant flowers with small or shallow cup-like coronas, 

the leaves are very different, usually grey-green and angled in cross-section 

with conspicuous keels on the underside, not green and terete as in the Jonquillae. 

They are generally small plants, not more than 20 cm tall and the flowers are only 

1.2–4 cm in diameter. There may be up to five flowers in an umbel but several taxa 

usually have solitary flowers. 

N. albimarginatus Muller-Doblies. Flowers one or two, golden yellow with a white 

rim to the corona. This species exhibits heterostyly: at anthesis all six stamens of 

the short-styled plants are exserted from the corona, unlike other species in the 

group (but N. gaditanus does have the upper three exserted). Morocco. 

N. atlanticus F.C. Stern. Flowers solitary, creamy-white, about 3.5 cm diameter; 

corona cup-shaped, 6 mm deep, 11 mm wide. Of unknown origin, presumed to be 

Morocco. 

N. calcicola Mendonça. Flowers up to five, yellow, 1.7–2.5 cm diameter; corona 

cup-shaped, 4–8 mm deep, 6–9 mm wide. Central Portugal. 

N. cuatrecasasii Casas, Laínz and Ruiz Rejón. Flowers one or two, yellow, 2.2–3cm 

diameter; corona cup-shaped, 3–6 mm deep, 8–10 mm wide. Southern Spain. This 

was known previously as N. rupicola ssp. pedunculatus. The variety segimonesis 

(Casas) Casas is a variant from southern Spain which is said to have slightly smaller 

flowers, but this is not consistent. 

N. rupicola Dufour. 

ssp. rupicola. Flowers solitary, yellow, 2–4 cm diameter; corona a wide funnel, 

2–6 mm deep, 7–10 mm wide. Central and southern Spain, central Portugal. 

ssp. marvieri ( Jah. and Maire) Maire and Weiller. Flowers solitary, yellow, 

about 3.5 cm diameter; corona a wide funnel, ca. 10 mm deep, 15 mm wide. 

Morocco. 

ssp. watieri (Maire) Maire and Weiller. Flowers solitary, white, about 3.5 cm 

diameter; corona a wide funnel, 2–6 mm deep, 7–10 mm wide. Morocco. 

N. scaberulus Henriques. Flowers up to four, yellow, 1.2–1.7 cm diameter; corona 

cup-shaped, 2–5 mm deep, 5–7 mm wide. North-central Portugal. 

1d(iii). Subsection Chloranthi D.A. Webb [Sect. Chloraster (Haworth) E. Dorda and 

F.J. Fernández Casas] 

Description: Autumn-flowering; flowers green throughout; corona very small, 

ca. 1 mm deep. 

Type species: N. viridiflorus Schousboe. 

N. viridiflorus Schousboe. Leaves very narrow, dark green, not present at flowering 

time. Flowers very odourous/fragrant, one-five in an umbel, about 2–2.5 cm in


diameter, deep green throughout; perianth tube 1–1.5 cm long, segments 

narrowly oblong, reflexed; corona a shallow cup only ca. 1 mm deep, 6-lobed at the 

margin. Southern Spain, Morocco. 

1e. Section Tapeinanthus (Herbert) Traub [Sect. Braxireon (Rafinesque) B. Valdes] 

Description: Leaves filiform. Flowers autumnal, usually solitary; perianth tube very 

short; perianth segments spreading; corona rudimentary to ± absent. 

Type species: N. cavanillesii A. Barra and G. López. 

N. cavanillesii A. Barra and G. López [syn. Tapeinanthus humilis (Cavanilles) Herbert]. 

Leaves one or two per bulb, present at flowering time or appearing later, filiform, 

green. Flowers autumnal, usually solitary, erect or suberect, 2–2.5 cm in diameter, 

yellow; corona absent or rudimentary. South-west Spain, Morocco. 

Note: Larger-flowered forms have been found in the Atlas Mountains of North 

Africa. 

2. Subgenus Hermione (Salisbury) Spach [syn. Sect. Tazettae De 

Candolle] 

Description: Flowers vernal or autumnal, usually several in an umbel; perianth tube 

usually long and slender; perianth segments well-developed, spreading or slightly 

reflexed; corona a shallow cup, sometimes ± absent. Filaments ± straight, shorter 

than anthers; anthers dorsifixed, included, or the upper three slightly exserted 

from the tube. Chromosome number x = 5 (2n = 10, 20, 30 or sometimes 22, 44). 

Type species: N. tazetta L. 

Two sections of subgenus Hermione are recognised, Hermione and Aurelia. 

2a. Section Hermione 

Description: Leaves flat. Flowers usually vernal, rarely autumnal, several in an 

umbel; perianth tube cylindrical/narrowly infundibuliform; perianth segments 

spreading; corona well developed, usually cup-shaped. 


Three subsections of section Hermione are recognised, Hermione, Angustifolii and 

Serotini. 

2a(i). Subsection Hermione 

Description: Leaves flat, to 2 cm wide. Flowers usually vernal, more rarely autumnal; 

corona cup-shaped, to 6 mm deep. 


Note: The taxa in this subsection, comprising the very fragrant, cluster-headed 

Tazetta and Paperwhite Narcissus species and their relatives, have been classified in 

very different ways. Some authors have regarded all those listed below as separate 

species, others have grouped them as subspecies or varieties of either N. tazetta 

(those with yellow or bicoloured flowers) or N. papyraceus (those with wholly white 

flowers), while others have ‘sunk’ some of the names altogether, without any

42 B. Mathew 

recognition whatsoever. Here, in order that interesting variations may not be 

overlooked, they have been retained with their specific names, and for convenience 

grouped in two series under the two ‘umbrella’ species, N. tazetta and N. papyraceus. 

Series A. Hermione (including series Luteiflorae Rouy). N. tazetta and its relatives – 

perianth segments white or yellow, corona yellow or orange. 

N. tazetta L. Leaves grey-green. Scape to 45 cm, flattened in section. Flowers up to 

15 in an umbel, to 4 cm diameter, fragrant; perianth tube 1–2 cm long; segments 

white; corona deep yellow or orange. Widespread in the Mediterranean region. 

Note: Some wild forms of N. tazetta, although predominantly winter/spring-flowering, 

sometimes flower in autumn in the wild but do not appear to do so consistently 

when introduced into cultivation. This species has a long history of cultivation and 

is naturalised widely from the Mediterranean to Japan; it is sometimes referred to 

as the Chinese Sacred Lily. 

The following have been variously recognised as variants of N. tazetta or as separate 

species: 

N. aureus Loiseleur. Leaves green. Perianth segments bright yellow; corona darker 

yellow-orange. A wholly yellow-flowered Tazetta, similar to the often-cultivated 

‘Soleil d’Or’ which may be a selection of it. South-west France, north-west Italy. 

N. bertolonii Parlatore. Leaves slightly greyish-green. Perianth segments pale to 

bright yellow; corona bright yellow to orange. Probably inseparable from N. aureus. 

N. canaliculatus hort. (N. tazetta ssp. lacticolor). Leaves narrow (to 5 mm), grey-green. 

Scape usually less than 15 cm. Perianth segments white, corona yellow. Origin 

unknown. 

N. corcyrensis (Herbert) Nyman. Leaves slightly grey-green. Perianth segments 

pale yellow, narrow and not overlapping at base, sometimes slightly reflexed; 

corona yellow or orange-yellow, conspicuously lobed at margin. Corfu, possibly 

also southern France, Italy and the Balkan Peninsula. 

N. cupularis (Salisbury) Schultes. Leaves markedly blue-grey-green. Perianth 

segments and corona pale to bright yellow. Probably inseparable from N. aureus. 

N. cypri Sweet. Leaves slightly grey-green. Flowers large (4–5 cm diameter) with 

white perianth segments and pale yellow corona, expanded at mouth. Cyprus. 

N. italicus Ker-Gawler. Leaves green. Flowers large (4–5 cm diameter) with 

perianth segments pale creamy-yellow; corona yellow. Southern France, Italy, 

Sicily, Corsica, Sardinia. 

N. ochroleucus Loiseleur. Leaves green. Scape subterete. Flowers 2.5–3.5 cm 

diameter; perianth segments white, corona pale lemon yellow. Similar to N. italicus 

and probably best regarded as a minor variant of it. Southern France. 

N. patulus Loiseleur. Leaves grey-green. Scape up to 20 cm. Flowers 1.8–2.5 cm 

diameter; perianth segments slightly reflexed, white; corona deep yellow. Southern 

France, Italy, Sicily, Sardinia, Corsica, Balkan Peninsula. 

Series B. Albiflorae Rouy. N. papyraceus and its relatives – perianth segments white, 

corona white. 

N. papyraceus Ker-Gawler. Leaves markedly grey-green. Scape to 50 cm, rarely 

more, compressed. Flowers 2.5–4 cm diameter, in umbels of up to 20, very fragrant;


perianth tube 1.2–2.5 cm long; segments and corona wholly white; corona cupshaped, 

usually 2–4 mm deep, slightly crenulate at the margin. South Europe 

and North Africa; widespread in the central and western Mediterranean region, 

westwards to Portugal. 

The following have been treated as variants of N. papyraceus or as separate 

species: 

N. barlae Parlatore. Leaves very grey-green. Flowers 2–2.5 cm diameter. Probably 

conspecific with N. panizzianus (see below). 

N. canariensis Burbidge. Leaves greyish-green. Flowers small (at most 1.5 cm 

diameter) with pointed petals. Canary Islands. 

N. pachybolbos Durieu. Bulb very large (5–7 cm diameter at maturity). Leaves pale 

grey-green. Flowers 1.5–2 cm diameter, corona less than a third as long as perianth 

segments. Algeria, Morocco. 

N. panizzianus Parlatore. Leaves grey-green. Scape very strongly compressed, 

two-edged. Flowers 2–2.5 cm diameter; perianth segments pointed. South-east 

France, Italy, Spain, Portugal. 

N. polyanthus Loiseleur. Leaves green. Scape terete. Flowers 2.5–4 cm diameter; 

corona entire. Southern France. 

The following two, although wholly white-flowered, appear to be sufficiently distinct 

from N. papyraceus and its relatives to merit specific status: 

N. dubius Gouan. Leaves dark green, slightly glaucous, 3–5 mm wide. Scape to 

20 cm. Flowers up to six in each umbel, 1.5–2 cm diameter; perianth tube 1–1.4 cm 

long; corona half as long as petals, ca. 4 mm deep. Southern France, north-eastern 

Spain. 

Note: Cytological investigations suggest that this has arisen by hybridisation 

between N. assoanus and N. papyraceus, but since it is now established over a wide 

area it is probably best regarded as a species in its own right. Although included 

here in subsection Hermione, there is a case for a separate subsection to house it. 

N. tortifolius Casas. Leaves grey-green, twisted lengthwise, 5–8 mm wide. Scape 

terete-elliptical in section, to 28 cm. Flowers up to 16 in each umbel, about 1.5 cm 

diameter; perianth tube ca. 10 mm long; corona ca. 2 mm deep. South-eastern 

Spain. 

2a(ii). Subsection Angustifolii (A. Fernandes) F.J. Fernández Casas 

Description: Leaves narrow (usually under 0.5 cm wide), flat or subterete. Flowers 

autumnal; corona very shallow, 2 mm or less deep. 

Type species: N. elegans (Haw.) Spach. 

N. elegans (Haw.) Spach. Scape ca. 20 cm. Leaves usually 2–5 mm wide, flattish 

or subterete, grey-green. Flowers autumnal, fragrant, up to seven in an umbel, 

2.5–3.5 cm in diameter; perianth segments white, cream or greenish-white; corona 

a shallow cup 1.5–2 mm deep with an incurved margin, green, greenish-brown 

or orange. West and south Italy, Sicily, Algeria, Libya, Morocco.

44 B. Mathew 

Note: N. elegans shows considerable infraspecific variation and some of the variants 

have been named: var. elegans forma elegans has narrow, pointed, white perianth 

segments and greenish corona; var. elegans forma auranticoronatus Maire has an 

orange corona; var. fallax Font-Quer has narrow, pointed, greenish-white perianth 

segments; var. flavescens Maire has narrow, pointed cream-coloured perianth 

segments; and var. intermedius J. Gay has perianth segments rather broader and 

more obtuse than in the other taxa. 

2a(iii). Subsection Serotini Parlatore 

Description: Leaves very narrow, ± filiform. Flowers autumnal, often solitary; perianth 

tube cylindrical; corona a very shallow cup. Filaments subequal to anthers, 

anthers dorsifixed. 

Type species: N. serotinus L. 

N. serotinus L. Scape 10–25 cm. Leaves absent at flowering time, filiform, green, to 

25 cm long. Flowers autumnal, fragrant, one to three per umbel, up to 3.5 cm in 

diameter, white; corona saucer-shaped, to 2 mm deep, orange or yellow; perianth 

tube 1.2–2 cm long, slender. Widespread in the Mediterranean region. 

Note: This is a variable species and some of the local variants have been given distinguishing 

names, but it is doubtful if they can be maintained. 

2b. Section Aurelia (J. Gay) Baker 

Description: Leaves flat. Flowers autumnal, several in an umbel; perianth tube 

cylindrical/narrowly infundibuliform; perianth segments spreading; corona rudimentary. 

Filaments longer than the anthers, anthers dorsifixed, exserted from the 

tube. 

Type species: N. broussonetii Lagasca. 

N. broussonetii Lagasca. Leaves present at flowering time, flat, grey-green, 1–1.5 cm 

wide. Flowers autumnal, fragrant, up to ten (rarely to 12) in an umbel, about 3 cm 

in diameter, white; corona represented by only a tiny rim, white. Morocco. 

Note: A large-flowered (about 3.5 cm diameter) tetraploid variant has been distinguished 

as forma grandiflorus. 

3. Subgenus Corbularia (Salisb.) Pax [Section Bulbocodium De Candolle] 

Description: Leaves narrow, semi-terete, usually dark or bright green, sometimes 

slightly glaucous. Flowers vernal (sometimes in late autumn or winter at low altitudes 

and in cultivation), solitary, usually held just below to just above the horizontal; 

perianth tube widely obconical; segments narrow, suberect or occasionally nearly 

patent; corona narrowly conical to widely funnel-shaped and constituting the 

main part of the flower. Filaments very much longer than the anthers, deflexed in 

the lower part, curved-ascending in the upper part, anthers dorsifixed, exserted from 

the tube, usually included within the corona but sometimes exserted. Chromosome 

number x = 7 (2n = 14 to 56).


Type species: N. bulbocodium L. 

Note: The ‘Hoop Petticoat’ daffodils cannot be confused with any other group of 

species, with their very narrow perianth segments and prominent, broadly funnelshaped 

corona. However, within the group they present great problems of classification. 

Many taxa have been described, at varying levels, but the account of 

Fernandes (1967) is that usually followed, recognising five species within the 

section – N. bulbocodium, N. romieuxii, N. cantabricus, N. obesus and N. hedraeanthus, 

with several named variants of each of the first three of these; this classification is 

largely followed here. Progress will probably only be made via a thorough field 

study of the plants comprising this section using modern statistical methods, 

coupled with molecular investigations. Although many of the taxa, as listed below, 

have characters which overlap those of others, thus rendering the treatment 

unsatisfactory, it would be even more unsatisfactory, especially for the purposes of 

communication, to lump all of them together under one species, N. bulbocodium. 

N. blancoi Barra and Lopez. This is the plant known previously as N. cantabricus 

ssp. luteolentus Barra and Lopez (see below). 

N. bulbocodium L. Flowers in varying shades of yellow. Anthers usually included 

or equalling the corona. 

ssp. bulbocodium Usually spring-flowering. Flowers pale to bright yellow, usually 

2.5–4.5 cm long; corona obconical, not narrowed at the mouth. 

var. bulbocodium. Pedicel usually up to 2 cm long; perianth usually up to 

3 cm long. Spain, Portugal, south-western and western France, Morocco. 

var. nivalis (Graells) Baker. Similar to above; abaxial surface of leaves 

deeply striate, scape ridged. Mountain plants from Spain, Portugal, Morocco. 

Plants described from Spain as N. jeanmonodii Casas, N. juressianus Casas and 

N. subnivalis Casas all appear to be very similar to this. 

var. quintanilhae Fernandes. This is described as large, 15–40 cm tall with 

flowers up to 3.5 cm long. Central-eastern Portugal. 

var. conspicuus (Haworth) Baker. Robust plants with leaves to 2.5mm 

wide. Pedicels usually more than 2 cm long. Flowers usually 3–3.5 cm 

long; corona ca. 2 cm diameter at the mouth. Spain, Portugal, western 

France. 

var. serotinus (Haworth) A. Fernandes. Similar to above but leaves to 

4 mm wide. Flower usually 3.5–5 cm long; corona ca. 3 cm diameter at mouth. 

Western Portugal. 

var. citrinus Baker [syn. N. lainzii Barra and Lopez]. Flowers lemon 

yellow, 3.5–5 cm long; corona ca. 2.5 cm diameter at the mouth, crenulate. 

Northern Spain. 

var. graellsii (Webb) Baker. Dwarf plants with primrose coloured flowers 

with exserted stamens. Central Spain. 

var. ectandrum Casas. Dwarf plants with prostrate leaves; flowers yellow 

with a widely flaring corona and spreading perianth segments. Central 

Spain. 

var. pallidus (Gatt. and Weiller) Maire and Weiller. Robust, with primrose 

yellow flowers; corona a wide, wavy-margined funnel ca. 3.5 cm across. 

Morocco.

46 B. Mathew 

ssp. praecox Gatt. and Weiller. Winter-flowering; flowers primrose 

yellow, 4.5–5 cm long; corona widely funnel-shaped, to 3.7 cm diameter. 

Morocco. 

var. praecox. Perianth segments with six veins. Morocco. 

var. paucinervius Maire. Perianth segments with three veins. Morocco. 

N. cantabricus DC. Flower pure white or greenish-white, usually produced in winter; 

anthers usually included or equalling the corona. 

ssp. cantabricus. Leaves usually more than one per bulb. 

var. cantabricus. Leaves usually two per bulb, spreading or prostrate. 

Flowers pure white. Spain. 

var. foliosus. Leaves three to eight per bulb, erect. Flowers milky-white, 

born on a distinct pedicel; corona 2–3cm diameter. Morocco. 

var. petunioides. Leaves one to three per bulb. Flowers white; corona up to 

4 cm diameter, very widely flared and markedly crenulate. Algeria. 

var. kesticus. Leaves usually two to four per bulb. Flowers milky- or greenishwhite; 

corona 2.5–3 cm diameter. Morocco. Plants described as N. peroccidentalis 

Casas from western Morocco are probably the same as this, with 

flowers often not pure white. 

ssp. luteolentus Barra and Lopez. This was described from south-east Spain 

(Albacete) and is said to have noticeably yellowish flowers and dark brown 

spathes. 

ssp. monophyllus (Dur.) A. Fernandes. Leaf one per bulb, prostrate. Flowers 

pure white; corona 1–2cm long, 2–3.5 cm diameter. South-eastern Spain, 

Morocco, Algeria. 

ssp. tananicus (Maire) A. Fernandes. Leaves erect, three to five per bulb. Flowers 

ca. 5 cm long with off-white corona and pale yellow perianth segments; corona 

narrowly conical, 2–2.5 cm diameter. Morocco. 

N. hedraeanthus (Webb and Heldr.) Colmeiro. Dwarf plant (scape 5–8cm) with 

small straw yellow flowers facing obliquely upwards or suberect; perianth segments 

wide (ca. 5 mm); stamens exserted from corona. South-eastern and south-central 

Spain. 

N. jacquemoudii. Casas is a little-known species from Morocco, said to resemble 

N. [bulbocodium var.] graellsii. 

N. obesus Salisbury. Leaves prostrate. Flowers large, usually deep yellow; corona 

with incurved margin. Western and southern Portugal, Morocco. 

N. romieuxii Braun Blanquet and Maire. Flowers rather pale, sulphur yellow to 

greenish-white; anthers usually exserted from corona. 

ssp. romieuxii. Flowers pale sulphur yellow. 

var. romieuxii. Pedicels absent or very short, to ca. 5 mm; flowers large, up 

to 4 cm long; perianth segments almost as long as corona. Morocco. 

Note: N. romieuxii var. mesatlanticus Maire is probably just one of the variants of 

var. romieuxii.


var. rifanus (Emb. and Maire) A. Fernandes. Pedicels to 1 cm long; flowers 

small, usually 2–3.5 cm long, petals longer than corona. Morocco. 

ssp. albidus (Emb. and Maire) A. Fernandes. Flowers white with a greenish or 

yellowish tint. 

var. albidus. Flowers suberect to erect, perianth segments distinctly longer 

than corona. Morocco. 

var. zaianicus (Maire, Weiller and Wilczek) A. Fernandes. Flowers near 

horizontal, perianth segments about as long as corona. Morocco. 

N. tingitanus. Casas, from Tangier, is described as having large pale yellow flowers 

becoming cream-coloured, then white with age. 

THE HORTICULTURAL CLASSIFICATION OF NARCISSUS CULTIVARS 

Historical developments 

The development of the horticultural classification of Narcissus cultivars was 

described by Kington (1998). Prior to 1884, garden varieties of daffodils were 

known by ‘pseudo-botanic names’, resulting in elaborate examples such as ‘Chalcedonicus 

Fimbriatus Multiplex Polyanthos’ and ‘Gallicus Major Flore Pleno’. In 

1884, the Royal Horticultural Society (RHS) Daffodil Conference reviewed the 

classification of garden varieties for the first time. Varieties were put into 33 

groups split between three divisions, the Magnicoronati, Mediicoronati and Parvicoronati. 

These groups and divisions were based partly on Baker’s (1869) 

classification of Narcissus species and partly on other criteria devised by the 

Conference Committee. While the group names were largely abandoned, elements 

of the 1884 classification – such as the use of both ‘arbitrary’ features (e.g., relative 

corona size) and ‘natural’ features (the characteristics of the species) – remain in 

the system of classification used today. Variety names were now in the vernacular, 

rather than in Latin, and some 400 were listed (Barr and Moore, 1884). With a 

great increase both in the numbers of varieties and in hybridisation between the 

three divisions, later RHS classification saw the three divisions amended to seven 

in 1908 and to 11 in 1910. The 1910 scheme introduced sub-divisions indicating 

colour for trumpet, large-cupped and small-cupped cultivars. The 1910 scheme 

survived, with only minor amendments, until 1950 (Kington, 1998). 

The revised classifications introduced by the RHS in 1950 and 1998 were 

designed to be more logical, easier to use, and adaptable to further developments 

in daffodil breeding (Kington, 1998). In 1950, the trumpet, large-cupped and 

small-cupped divisions were sub-divided according to a one-letter colour code: 

(a) perianth and corona coloured, (b) perianth white, corona coloured, (c) perianth 

and corona white, and (d) other combinations. A more comprehensive colour coding 

was introduced in 1975. Main changes in 1998 included the sub-division of splitcorona 

cultivars into Collar and Papillon types, and the establishment of a division 

for Bulbocodium cultivars. With the huge number of hybrids and intense interest 

among enthusiasts, it is likely that the classification of daffodil cultivars will continue 

to evolve.

(1) (2) 

(3) (4)

(5) (6) 

The present classification 


Figure 3.2 Examples of Narcissus cultivars of the main cultivar groups, with their classification: 

(1) Small-cup: ‘Barrett Browning’, 3 WWY-O; (2) Large-cup: ‘Hollywood’, 

2 Y-O; (3) Trumpet: ‘Dutch Master’, 1 Y-Y; (4) Double: ‘Papua’, 4 Y-Y; (5) Tazetta: 

‘Grand Soleil d’Or’, 8 Y-O; (6) Split-corona: ‘Canasta’, 11a W-Y. Photos: Horticulture 

Research International. (See Colour plate 2) 

The RHS 1998 scheme of classification into 13 divisions is shown in Table 3.1 

(Kington, 1998). Once a selection has been distinguished by a cultivar name, 

whether of cultivated or wild origin, it is placed in one of twelve divisions, division 

13 being reserved for those Narcissus (including hybrids) distinguished solely by a 

botanical name. The majority of large-flowered cultivars fall into divisions 1 to 3, 

the trumpet, large-cupped and small-cupped varieties, respectively, the separation 

depending on the ratio of the lengths of perianth segments and corona. Other distinct 

flower types are the double cultivars (division 4) and the split corona cultivars 

(division 11), the latter being classified as Collar Daffodils (11a) or Papillon Daffodils 

(11b) on the basis of the corona and perianth segments being opposite or 

alternate, respectively. Cultivars with clearly evident characteristics of Narcissus 

triandrus, N. cyclamineus, section Jonquilla or Apodanthi, section Tazettae, the 

N. poeticus group and section Bulbocodium fall into divisions 5 to 10, respectively. 

Division 12 is reserved for those cultivars which do not fit the description of any 

other division. Since 1975, a more comprehensive system of defining flower colour 

has been used, the colour code being appended to the division number. The present

50 B. Mathew 

Table 3.1 Horticultural classification of Narcissus cultivars a 

Division 

1 Trumpet daffodils Corona (‘trumpet’) as long or longer 

than perianth segments 

2 Large-cupped daffodils Corona (‘cup’) more than one-third the 

length of the perianth segments, but 

not as long 

3 Small-cupped daffodils Corona (‘cup’) not more than one-third 

the length of the perianth segments 

4 Double daffodils Corona and (or) perianth segments with 

doubling 

5 Triandrus daffodils Characteristics of N. triandrus clearly 

evident 

6 Cyclamineus daffodils Characteristics of N. cyclamineus clearly 

Note 

a after Kington (1998). 

scheme describes the colour (White, Green, Yellow, Pink, Orange or Red) of the 

perianth segments followed by that of the corona. Either component may have a 

single-letter colour code if substantially of one colour, or a three-letter code describing 

the colour of the distal, mid and proximal regions of the perianth segments 

or of the proximal, mid and distal regions of the corona, respectively. Full 

details of these criteria are given by Kington (1998). Some cultivars representing 

these divisions and classifications are shown in Figure 3.2. 

The classified lists 

evident 

7 Jonquilla and Apodanthus Characteristics of Sections Jonquillae or 

daffodils 

Apodanthi clearly evident 

8 Tazetta daffodils Characteristics of Section Tazettae clearly 

evident 

9 Poeticus daffodils Characteristics of N. poeticus group 

10 Bulbocodium daffodils Characteristics of Section Bulbocodium 

11 Split corona daffodils: 

(a) Collar daffodils 

(b) Papillon daffodils 

clearly evident 

Corona split: 

with corona segments opposite the 

perianth segments 

with corona segments alternate to the 

perianth segments 

12 Other daffodils Those that do not fit descriptions of 

any other division 

13 Daffodils distinguished solely 

by botanical name 

The first RHS list of daffodil cultivars, including about 1500 names, appeared in 

1907 (RHS, 1907), followed by a series of Classified Lists beginning in 1908 (RHS, 

1908) and, following the appointment of the RHS as the International Registration 

Authority for Narcissus varieties in 1955, a series entitled Classified List and International 

Register starting in 1958 (RHS, 1958). The International Daffodil Checklist 

of 1989 (Kington, 1989) preceded publication in 1998 of the 23rd cumulative list


of names as The International Daffodil Register and Classified List (Kington, 

1998), containing about 25 000 distinct names after accounting for synonymy 

(S. Kington, personal communication). In order to avoid confusion due to the 

re-use of earlier names, from 1998 the Register re-instated a quantity of names 

that had previously been deleted because they had been deemed to be extinct or 

of no historical interest. The Register is updated annually by supplements of 

newly registered names (RHS, 1999 and earlier years). A further advantage of the 

current series (1998 Register onwards) is that parentages and descriptions are 

being included where available. 

ACKNOWLEDGEMENTS 

The section on the horticultural classification of narcissus cultivars was contributed 

by Gordon Hanks with the help of Sally Kington (Daffodil Registrar, Royal Horticultural 

Society). 

REFERENCES 

Baker, J.G. (1869) Review of the genus Narcissus. Gardeners’ Chronicle, 416, 529, 686, 1015, 

1136, 1183. 

Baker, J.G. (1875) Review of the genus Narcissus. In: F.W. Burbidge (ed.), The Narcissus: its 

History and Culture, Reeve, London. 

Barr, P. and Moore, T. (1884) Nomenclature of Narcissus. Gardeners’ Chronicle, 21, 607–608. 

Blanchard, J.W. (1990) A Guide to Wild Daffodils. Alpine Garden Society, Woking. 

Brandham, P.E. (1992) Chromosome numbers in Narcissus cultivars and their significance 

to the plant breeder. The Plantsman, 14, 133–168. 

Brandham, P.E. and Kirton, P. (1987) The chromosomes of species, hybrids and cultivars of 

Narcissus L. Kew Bulletin, 42, 65–102. 

Cullen, J. (1986) Narcissus. In: S.M. Walters, A. Brady, C.D. Brickell, J. Cullen, P.S. Green, 

J. Lewis, V.A. Matthews, D.A. Webb, P.F. Yeo and J.C.M. Alexander, The European Garden 

Flora, Vol. 1, Cambridge University Press, Cambridge, pp. 301–309. 

Fernandes, A. (1951) Sur la phylogénie des espèces du genre Narcissus L. Boletim da 

Sociedade Broteriana series 2, 25, 113–190. 

Fernandes, A. (1967) Contribution à la connaissance de la biosystématique de quelques 

espèces du genre Narcissus L. Potugaliae Acta Biologica, 9, 1–44. 

Fernandes, A. (1969a) Contribution to the knowledge of the biosystematics of some species of Genus 

Narcissus L. 5th Simposio de Flora Europaea, Trabajos y Comunicaciones, Sevilla, 

pp. 245–284. 

Fernandes, A. (1969b) Keys to the identification of native and naturalized taxa of the genus 

Narcissus L. Daffodil and Tulip Year Book 1968, pp. 37–66. 

Haworth, A.H. (1831) Narcissinearum Monographia. Ridgway, London. 

Herbert, W. (1837) Amaryllidaceae. Ridgway, London. 

Kington, S. (1989) The International Daffodil Checklist. Royal Horticultural Society, London. 

Kington, S. (1998) The International Daffodil Register and Classified List 1998. Royal Horticultural 

Society, London. 

RHS (1907) List of Daffodil Names. Royal Horticultural Society, London. 

RHS (1908) The Classified List of Daffodil Names. Royal Horticultural Society, London.

52 B. Mathew 

RHS (1958) The Classified List and International Register of Daffodil Names. Royal Horticultural 


RHS (1999) The International Daffodil Register and Classified List 1998. Second Supplement. 

Royal Horticultural Society, London. 

Spach, E. (1846) Histoire Naturelle des Végétaux Phanerogames, Vol. 12. Librairie encyclopédique 

de Roret, Paris. 

Webb, D.A. (1980) Narcissus. In: T.G. Tutin, V.H. Heywood, N.A. Burges, D.M. Moore, 

D.H. Valentine, S.M. Walters, D.A. Webb, A.O. Chater and I.B.K. Richardson, Flora Europaea, 

Vol. 5, Cambridge University Press, Cambridge, pp. 78–84. 

ADDITIONAL BIBLIOGRAPHY 

Baker, J.G. (1888) Handbook of the Amaryllideae. George Bell, London. 

Bowles, E.A. (1934) A Handbook of Narcissus. Hopkinson, London. 

Fernandez Casas, J. (1981) Many publications on Narcissus in Fontqueria Vol. 1 onwards. 

Jefferson-Brown, M. (1991) Narcissus. Batsford, London. 

Meyer, F.G. (1966) Narcissus species and wild hybrids. American Horticultural Magazine, 45, 

47–76. 

Wells, J.S. (1989) Modern Miniature Daffodils. Batsford, London.

4 Commercial production of Narcissus 

bulbs 


INTRODUCTION 

Narcissus (daffodil) bulbs have been an important floricultural crop in western 

Europe since the late nineteenth century, although the bulbs have been grown in 

the Netherlands since the sixteenth century and Narcissus hispanicus has been 

cultivated in the UK for over 300 years as ‘N. maximus’ or ‘N. maximum superbus’ 

(Doorenbos, 1954). In a 1998 survey of consumers in the UK, daffodils were rated 

eighth in popularity amongst cut-flowers and achieved sixth position in value of 

sales, despite being relatively inexpensive and not available throughout the year 

(FPA, 1999). At the start of the twenty-first century, the narcissus or daffodil 

remains one of the major ornamental bulb crops grown in temperate regions, with 

large areas of field-grown crops providing both bulbs and flowers, while bulb 

‘forcing’ in glasshouses provides flowers and pot-plants over an extended season. 

Rees (1993) estimated that the area of narcissus grown in gardens, parks, cemeteries, 

etc., is five-times the area grown commercially. The histories of commercial bulb 

growing in the UK, Netherlands and US, the major producing countries, have 

been described by Dobbs (1983), Krelage (1946) and Gould (1993), respectively. 

After a brief review of the statistics of narcissus bulb production, this chapter will 

describe the methods used in growing the crop. The objective is to provide clear 

guidance on how narcissus are grown, commenting on how the requirements of 

producing bulbs for processing might differ from the production of bulbs as 

ornamentals. Research findings will also be considered, for these may give insight 

into how the methods of production of narcissus as industrial crops might be 

varied or improved. 

PRODUCTION STATISTICS 

World production of Narcissus bulbs 

The areas of field-grown narcissus in the major producing countries are given in 

Table 4.1. Production is dominated by the UK and Netherlands, with some 4200 

and 1800ha, respectively, although it should be noted that in the UK the crop is 

grown on a two-year-down basis so that only half the area is lifted each year. 

It should also be noted that, since 1996–1997, the practice in the UK has been to 

include statistics on narcissus grown for ornamental use only, and examination of 

recent figures (MAFF, 1999a) suggests that a few hundred hectares have been

54 G.R. Hanks 

grown for galanthamine production. Over 400ha are grown in the USA, and other 

significant areas of bulb production include Australia and Canada (British Columbia). 

Significant areas are also grown in Jersey (Channel Islands), although this is 

mainly for flower, not ‘dry bulb’, production. Tazetta narcissus bulbs are produced 

mainly in Israel. In general, comparisons of areas of the field-grown crops over 

the past 10 years show that production areas are generally stable, with a small 

increase (Table 4.1). However, recent trends include a significant increase in the 

area grown in the Republic of Ireland, while in Poland the area has decreased 

markedly, largely due to virus infection (Mynett, 1990). 

The annual output of narcissus bulbs can be estimated from the area lifted annually, 

the average planting density (say about 17.5 t/ha), and the average percentage 

Table 4.1 World production areas of field-grown Narcissus 

Notes 

a 1980s figures from Hanks (1993). 

b Excludes area grown for processing. 

c Mainly for flower production. 

na: not available. 

1980s a 1990s 

Year Area (ha) Year Area (ha) Reference 

Australia – na 1999 200 G. Guy (personal communication) 

Canada (British 

Columbia) 

– na 1991 149 Gould (1993) 

Denmark 1982 36 – na 

England and Wales 1990 3972 1998 b 3808 MAFF (1999a) 

France 1981 21 1999 22 M. le Nard (personal 

communication) 

Germany c 1984 29 1996 14 Heinrichs (1999) 

Republic of Ireland 1985 26 1991 73 Heinrichs (1999) 

Israel 1987 144 1999 150 G. Lurie and H. Lilien-Kipnis 

(personal communication) 

Italy c 1982 80 1994 27 Heinrichs (1999) 

Japan 1987 28 1997 44 Japanese Ministry of Agriculture 

and Fisheries (K. Ohkawa, 

personal communication) 

Jersey c 1990 298 1998 175 Jersey Department of Agriculture 

and Fisheries (I.K. Norris, 

personal communication) 

Netherlands 1990 1639 1998 1756 PT/BKD (1999) 

New Zealand – na 1999 70 J. Catley (personal communication) 

Northern Ireland – na 1996 4 Department of Agriculture 

Northern Ireland (personal 


Poland 1982 200 1999 50 D. Sochacki and K. Mynett 

(personal communication) 

Scotland 1987 255 1997 390 M.W. Sutton (personal 


South Africa – na 1992 12 De Hertogh et al. (1992) 

USA 1989 467 1998 410 G.A. Chastagner (personal 


Total of above 7195 7354

Production of Narcissus bulbs 55 

Table 4.2 Areas (ha) of field-grown Narcissus in the UK and the Netherlands 

1988 a 1989 1990 1991 1992 1993 1994 1995 1996 1997 1998 

UK b 3826 3961 3972 3702 3943 4048 3830 4334 3993 3936 3808 

Netherlands c 1763 1799 1652 1533 1343 1350 1406 1496 1517 1574 1756 

Notes 

a 1988 means 1988–1989, etc. 

b MAFF (1999a), 1998 figure provisional. 

c PVS/BKD (1993), PT/BKD (1999), 1998 figure provisional. 

weight increase from planting to lifting (say 150%); disposable yield is the yield 

after taking out the weight required as re-planting stock. This gives an annual disposable 

bulb output of about 30 000 tonnes for both the UK and the Netherlands. 

A tonne of narcissus bulbs typically contains about 20 000 ‘medium sized’ bulbs. 

Narcissus bulbs are sold as commercial planting stocks, for commercial bulb 

forcing, for amenity or landscape use, and for retail sales either loose or in ‘prepacks’, 

the last involving garden centres, multiple retailers, mail order and other 

outlets. In England and Wales, commercial bulb forcing currently accounts for 

over 4000 tonnes of narcissus bulbs (about 80 million bulbs) annually (MAFF, 

1999b). The prices of narcissus bulbs may change markedly over periods of a few 

years, and there is an interplay between the price of bulbs and flowers, the areas of 

field-grown crops from which flower crops are taken, and the number of bulbs 

used for forcing. In the UK, recent trends have increased the significance of cutflowers 

taken from field-grown crops and have resulted in fewer bulbs being 

forced. When bulb prices are favourable, bulbs used for forcing may be re-claimed 

and planted back in the field. Bouwman (1993) described a simulation model for 

the supply and demand of bulbs: on the demand side there was a relationship 

between price asked and quantity demanded, and, on the supply side, between 

price asked and acreage grown. The economic aspects of narcissus bulb production 

are considered in Chapter 5 of this volume. 

Bulb growing areas in the main producing countries 

In England and Wales, some two-thirds of the field-grown narcissus are grown in 

eastern England (Lincolnshire, Norfolk and Cambridgeshire) and much of the 

remainder in the south-west, in Cornwall and the Isles of Scilly. Census figures for 

1998 give the following areas for bulb and outdoor flower crops (predominantly 

narcissus) of 2016 ha (south Lincolnshire), 463 ha (Norfolk), 364 ha (Cambridgeshire) 

and 1472 ha (Cornwall and Isles of Scilly), out of a total of 5325 ha (MAFF 

Statistics Branch, personal communication). For narcissus alone, it is now (1999) 

considered that some 50% of the production in England and Wales is in the eastern 

counties, and 40% in the south-west, with a gradual shift towards the latter 

(J.B. Briggs, personal communication). South Lincolnshire is an area with strong 

historical links to the Netherlands, evidenced, for example, by the district name 

‘South Holland’. The area is characterised by large fields, extensively mechanised 

agriculture, and the efficient production of cereals, potatoes and vegetables as well 

as narcissus bulbs, a logical development as there is scope for sharing equipment

56 G.R. Hanks 

such as drying stores with potato, onion or cereal growing (e.g., see Williams, 1996). 

Until recent years, flowers cropped from the field were considered of secondary 

importance to the bulbs. This area has traditionally supplied the relatively close 

markets of London and the industrial Midlands, and is now a major distribution 

centre for fresh produce for the UK, the EU and world-wide. Cornwall has been 

traditionally important for the production of early fresh produce, including 

flowers and potatoes, from its mild, relatively frost-free climate. With improved 

road links eastwards, and EU support, the area has become increasingly important 

for narcissus bulb production. Bulb growing has a very high labor requirement 

for flower picking and bulb handling, and so is important in the local economy. 

Bulb quality is enhanced by soils free of potato cyst nematode, a requirement for 

bulbs exported to the USA, Canada and Norway. Unlike eastern England, fields 

are sometimes relatively small, of irregular topography and have stony soils. The 

extreme of this type of agriculture is found in the Isles of Scilly, where Tazetta 

narcissus are cropped over a long season in the frost-free climate: fields are small 

and often surrounded by hedges or artificial shelter for protection from the 

strong, salt-laden winds. The high transport costs here limit growing to high-value 

produce, and there is an interdependence between agriculture and tourism. 

Other areas of narcissus growing include Scotland and Jersey, the latter mainly for 

flower production. Growing narcissus over the broad geographic range of the 

British Isles ensures flower production over a long season and confers a variety of 

other climatic advantages, such as frost-freedom in the extreme south-west, and 

relative freedom from insect pests in the colder north. 

In the Netherlands, bulbs have been traditionally grown around Hillegom, in the 

Bloembollenstreek. The advantages to bulb production here are considerable: ideal 

sandy soils with a controlled water table, abundant traditional expertise, a strong 

R&D base, the availability of family labour for labour-intensive bulb handling, and 

a superb infrastructure for marketing, selling and logistics. With the pressure on 

agricultural land in this region in recent years, however, much narcissus production 

has moved to the heavier soils of the polders of North Holland, more akin to 

the situation in eastern England. About 55% of the production area is in Noord- 

Holland and 33% in the Bloembollenstreek (de Vroomen and de Groot, 1991). 

In USA, the most commercial bulb production takes place in the ideal climatic 

conditions of the coastal area of the Pacific North-West, with almost all narcissus 

production in Washington State. Most growers now produce field-grown flowers 

as well as bulbs, although formerly the production of bulbs for glasshouse forcing 

in the east and mid-west was important. The tourism associated with bulbs has a 

major impact in the communities where bulbs are grown, with a number of bulb 

festivals: it is estimated that the economic input of bulb growing on tourism is fivetimes 

the value of the crop itself (G.A. Chastagner, personal communication). 

Exports and imports 

Dutch exports and imports of narcissus bulbs are shown in Table 4.3 (although the 

words ‘exports’ and ‘imports’ are used here in a general sense, since the advent in 

the EU of the Single Market, the terms strictly apply only to trade with non-EU 

(‘third’) countries). The predominant Dutch export destinations are Germany, 

North America, the UK and France. Dutch imports of narcissus bulbs are dominated

Table 4.3 Dutch exports and imports of Narcissus bulbs (1990 figures) a 

Note 

a From PVS (1990a,b). 


by supplies from the UK, along with Tazetta bulbs from Israel (PVS, 1990a,b). 

The trade in narcissus bulbs between the UK and the Netherlands consists largely 

of bulbs of ‘mainstream’ cultivars (sold by weight) exported from the UK to the 

Netherlands, and bulbs of ‘choicer’ cultivars (sold by number) traded in the opposite 

direction. UK also exports bulbs to Germany, North America, Scandinavia and 

France. UK exports of bulbs (predominantly narcissus) were valued at £9.451 m in 

1998, while imports of narcissus bulbs amounted to £3.462 m (MAFF, 1999a). 

Israel exports some 30 million Tazetta bulbs annually (I. Gerstein and H. Lilien- 

Kipnis, personal communication), to the Netherlands and the USA, while China 

and Japan export small quantities to the USA (Oldfield, 1989). US imports about 

120 million narcissus bulbs annually (Gould, 1993), including some 90 million 

from the Netherlands (1994–1995 figures, W.B. Miller, personal communication). 

The Netherlands and UK also export large volumes of narcissus cut-flowers. On 

the assumption that narcissus make up the bulk of the cut-flower exports from the 

UK, these were valued at over £10m in 1998 (MAFF, 1999a). Pot-grown dwarf 

narcissus are an important export from the Netherlands: sales of pot narcissus in 

Germany reached 1.79 million in 1998 (Heinricks, 1999). 

Cultivars grown 

Exports Imports 

Number (million) Weight (1000 t) Number (million) Weight (1000 t) 

Austria 9 0.7 0 0 

Belgium and 

Luxembourg 

6 0.4 0 0 

Canada 11 0.6 0 0 

Denmark 8 0.4 0 0 

Finland 7 0.6 0 0 

France 29 1.6 8 0.3 

Germany 88 5.6 4 0.2 

Israel 0 0 11 0.5 

Italy 4 0.2 0 0 

Norway 4 0.3 0 0 

Sweden 13 1.0 0 0 

Switzerland 8 0.5 0 0 

UK 37 1.3 50 2.8 

USA 70 4.0 0 0 

Total 294 17.2 73 3.8 

Full statistics for the cultivars grown are available only for the Netherlands, and 

they are summarised in Table 4.4 by cultivar groups and in Table 4.5 by top 

cultivars. In recent years, the main change has been the expansion in growing 

Cyclamineus types (now 30% of the total narcissus area), predominantly involving

Table 4.4 Areas of Narcissus cultivar groups grown in the Netherlands in 1990–1991 and 

1998–1999 a 

Cultivar group 1990–1991 1998–1999 Change in area 

over 8 year period 

Area (ha) (No. of Area (ha) (No. of 

cultivars) 

cultivars) 

Cyclamineus 185 (25) 535 (32) +189% 

Large-cup 666 (107) 433 (164) –35% 

Trumpet (yellow) 375 (40) 294 (61) –22% 

Double 189 (40) 202 (58) +7% 

Tazetta 58 (10) 73 (17) +26% 

Jonquilla 20 (18) 53 (26) +165% 

Triandrus 15 (9) 34 (12) +127% 

Split Corona 30 (33) 25 (43) –17% 

Trumpet (bicolor) 27 (15) 19 (16) –30% 

Small-cup 34 (14) 18 (27) –47% 

Species 6 (10) 17 (10) +183% 

Trumpet (white) 16 (7) 13 (12) –19% 

Poeticus 6 (3) 6 (8) 0% 

Unknown 9 (–) 33 (–) – 

Totals 1639 (331) 1756 (488) b +7% 

Notes 

a Data from PVS/BKD (1991) and PT/BKD (1999); data or cultivars grown by only one grower are excluded. 

b Numbers do not total due to some un-classified cultivars. 

Table 4.5 Areas of the ‘Top 20’ Narcissus cultivars 

grown in the Netherlands in 1998–1999 a 

Cultivar Division and colour code b Area (ha) 

Tête-à-Tête 12Y-Y 449 

Carlton 2Y-Y 153 

Dutch Master 1Y-Y 135 

Ice Follies 2W-W 101 

Golden Harvest 1Y-Y 55 

Minnow 8Y-Y 46 

Dick Wilden 4Y-Y 45 

February Gold 6Y-Y 33 

Standard Value 1Y-Y 32 

Jetfire 6Y-O 30 

Salome 2W-PPY 28 

Tahiti 4Y-O 24 

Bridal Crown 4W-Y 22 

Gigantic Star 2Y-Y 19 

Quail 7Y-Y 18 

Van Sion 4Y-Y 16 

Hawera 5Y-Y 15 

Cheerfulness 4W-Y 15 

Geranium 8W-O 15 

Yellow Sun 2Y-Y 13 

Notes 

a From PT/BKD (1999). 

b Kington (1998).


‘Tête-à-Tête’, a dwarf cultivar important as a scented pot-grown bulb and for 

garden use. There have also been significant increases in the area of other more 

specialist types (Tazetta, Jonquilla, Triandrus and species), while the area of 

trumpet, large-cup and small-cup cultivars has declined, including once popular 

cultivars such as ‘Golden Harvest’. Examination of the figures in Table 4.4 shows a 

great increase in the number of different cultivars grown in the Netherlands, even 

in cultivar groups of which the overall area is declining. This may indicate the 

development of more discerning consumer tastes. In 1998, 488 cultivars were 

listed (those cultivars grown by a single grower were excluded from these statistics), 

of which the top five cultivars (‘Tête-à-Tête ’, ‘Carlton’, ‘Dutch Master’, ‘Ice 

Follies’ and ‘Golden Harvest’) made up 51% of the total area (PT/BKD, 1999). In 

the UK, one publication listed 35 cultivars as ‘important’ (ADAS, 1985a); the main 

ones included ‘Golden Harvest’, ‘Dutch Master’, ‘Carlton’, ‘Fortune’, ‘Ice Follies’ 

and ‘Cheerfulness’. However, there is now a trend away from ‘Golden Harvest’ 

and ‘Carlton’, which are susceptible to base rot disease. 

BULB PRODUCTION 

As the UK and the Netherlands account for some 76% of the world area of fieldgrown 

Narcissus bulbs, the information on bulb production methods in the present 

chapter is largely derived from British and Dutch advisory material for bulb 

growers. UK advisory material formerly included a range of booklets produced by 

the Ministry of Agriculture, Fisheries and Food (MAFF) and MAFF’s former Agricultural 

Development and Advisory Service (ADAS), in particular ADAS (1985a). 

Dutch material includes the handbook produced by the Ministerie van Landbouw, 

Natuurbeheer en Visserij (Ministry of Agriculture, Nature Management and Fisheries) 

with the Consulentschap in Algemene Dienst voor de Bloembollenteelt (General 

Service Consultancy Unit for Flower-bulb Cultivation) (Langeslag, 1990). Neither 

of these publications has been updated. Information quoted in this account without 

specific reference citations is taken from these two publications. Other useful 

accounts of commercial narcissus growing include: from the USA, NBGA (1957, 

1961); from the Netherlands, Krabbendam and Baardse (1964); and, from the UK, 

ADAS (1970) and MAFF (1984). In growing bulbs as ornamental crops, either the 

bulbs or the flowers, or both, may be important, depending on local circumstances 

and the economic situation. In growing bulbs for pharmaceutical or other industrial 

uses, quite different considerations may apply, and it may be appropriate to 

modify agricultural practices accordingly. More specialised protocols may be 

needed, at least in the initial stages of bulking suitable stocks or screening cultivars 

for potential use. On the other hand, where the aim is industrial processing, 

flower production and the visual aspects of bulb quality may be unimportant. 

Rees (1972) described the annual cycle of narcissus growth as characterised 

by alternating periods of growth and quiescence, which enable the genus in its 

natural, typically Mediterranean, habitat to condense its above-ground growth 

into the relatively short period between cool winters and hot, dry summers. This 

provides horticulturists with an annual opportunity to treat, grade and market the 

bulbs during the ‘dormant’ summer period (during which there is, in fact, active 

formation and growth of the young bulbs, shoots and root initials within the bulb).

60 G.R. Hanks 

Following planting in late-summer or autumn, root outgrowth is rapid, and shoot 

growth continues inside the bulb until slowed by falling temperatures. Except for 

Tazetta narcissus, the plants have a cold requirement for rapid, synchronous shoot 

growth and anthesis, and, once this requirement has been met by normal winter 

temperatures, shoots grow at a rate determined by ambient temperatures. Bulb 

growth is rapid from around the time of anthesis, but is soon curtailed by the 

prompt onset of foliar senescence in summer, perhaps as a means of conserving 

water. In the UK, initiation of the flower begins in May and its differentiation is 

completed in July or August. Further details of the annual pattern of narcissus 

growth are given in Chapter 1 of this volume. 

PESTS, DISEASES AND DISORDERS 

In the commercial production of narcissus bulbs, considerable effort is needed to 

control pests and diseases. As for other crops, pesticide applications in the field are 

important, but bulbs also present a convenient opportunity for pesticide applications 

during the ‘dormant’ stage between bulb lifting and re-planting. Many pests 

and diseases are exacerbated by the techniques of modern husbandry, such as 

high planting densities, reduced sorting of bulbs by hand, bulk handling and 

two-year-down growing (Price, 1977a,b), but, since it would be uneconomic to 

change these practices, control measures have to be highly effective to work under 

these exacting conditions. 

Methods of control (or management) of narcissus pests and diseases, and for 

preventing physiological disorders, are incorporated into the following description 

of narcissus bulb production. Further descriptions and methods of control of 

pests, diseases and disorders are available in a number of texts. For the UK, information 

on pests is given in Lane (1984), on diseases and disorders in Moore et al. 

(1979) and ADAS (1986a), and generally in Rees (1972, 1992), Linfield and Cole 

(1989), Hanks (1993) and Linfield (1994). Dutch information includes Bergman 

et al. (1978) and Langeslag (1990), and US information includes Gould and Byther 

(1979) and Chastagner and Byther (1985). 

Narcissus bulbs are unusual in that the major pest, stem nematode (‘eelworm’, 

Ditylenchus dipsaci), is controlled by immersing bulbs in a hot-water treatment 

(HWT). Other pests are also controlled by HWT, including the larvae of large 

narcissus fly (Merodon equestris), small narcissus flies (Eumerus strigatus and E. tuberculatus), 

other nematodes, narcissus leaf miner (Norellia spinipes), bulb scale mite 

(Steneotarsonemus laticeps) and bulb mites (Rhizoglyphus and Histiostoma species). Of 

these, the large narcissus fly and bulb scale mite are the most significant pests, 

while small narcissus flies and bulb mites attack only damaged bulbs. Besides 

D. dipsaci, other nematode pests include the narcissus bulb and leaf nematode 

(Aphelenchoides subtenuis) and the root-lesion nematode Pratylenchus penetrans that 

can attack bulbs causing a root rot in conjunction with the fungus Nectria radicicola. 

Nematodes transmitting narcissus viruses are Trichodorus and Paratrichodorus spp. 

(transmitting tobacco rattle virus), Longidorus spp. (tomato black ring and raspberry 

ringspot viruses) and Xiphinema diversicaudatum (arabis mosaic and strawberry 

latent ringspot viruses). Potato cyst nematode (PCN, Globodera spp.) does not 

attack narcissus bulbs, but its presence may cause the rejection of bulbs for export


to some countries. Slugs attack bulbs and flowers, and the garden swift moth 

(Hepialus lupulinus) is an occasional pest. Aphids do not usually colonise narcissus, 

but several common species spread viruses through exploratory probings. Vertebrate 

pests are not usually troublesome in narcissus growing. 

Narcissus fungal diseases include bulb rots and foliar diseases, of which the most 

significant is base rot (basal rot) caused by Fusarium oxysporum f.sp. narcissi. Many of 

the procedures in narcissus growing are designed to control base rot, although 

incidentally helping to control other fungal pathogens. The control of base rot 

involves a range of measures, of which the use of fungicides is only one (Melville, 

1980; Tompsett, 1980a; ADAS, 1989b). Some of the most widely grown narcissus 

cultivars, such as ‘Golden Harvest’ and ‘Carlton’, are susceptible to base rot. Apart 

from base rot (Tompsett, 1986), relatively little is known of varietal differences in 

susceptibility to other diseases and pests, although Beaumont (1950) gave some 

useful information. Neck rot is related to base rot, and is also generally caused by 

F. oxysporum f.sp. narcissi, although Penicillium hirsutum and Botrytis narcissicola are 

also implicated (Davies et al., 1998; Carder, 1999). Although reported by Hawker 

in 1935, neck rot has caused concern recently as it is seen in pre-export inspections, 

and it attacks a wider range of cultivars than base rot (Davies et al., 1998). 

It is important to distinguish pathological neck rot, which is a rot clearly spreading 

down from the bulb neck, from ‘physiological neck rot’, which is simply the 

natural presence of the dead bases of leaves in the bulb neck. Less usual, or less 

serious, fungal rots include grey bulb rot (Rhizoctonia tuliparum), soft rot (Rhizopus 

spp.), Penicillium rots, black slime (Sclerotinia bulborum) and white root rot (Rosellinia 

necatrix). The major fungal foliar diseases are smoulder (Botrytis narcissicola), 

fire (Sclerotinia polyblastis), leaf scorch (Stagonospora curtisii) and white mould (Ramularia 

vallisumbrosae). A number of fungi attacks narcissus bulbs resulting in skin 

diseases that give a downgraded appearance, with darker, greasy, multi-layered or 

irregular skins (Bergman et al., 1978). 

Bacterial diseases are not generally associated with narcissus. Although mycoplasma-like 

organisms have been reported in unhealthy narcissus plants (Bellardi 

et al., 1990), there is no information as to the wider significance of this discovery. On 

the other hand, some 21 viruses are known to infect narcissus, of which about 13 are 

considered to be of economic importance, especially the aphid-borne narcissus 

yellow stripe, narcissus late seasons yellows and narcissus white (or silver) streak 

viruses (Brunt, 1995). Most have aphid or nematode vectors, although for some the 

vector is not known or mechanical transmission has been demonstrated. Aphid- and 

nematode-borne viruses are unlikely to be spread by handling healthy and infected 

plants alternately, and Mowat (1980a) found no evidence for the mechanical transmission 

of narcissus tip necrosis virus by handling and flower cropping. However, 

narcissus mosaic virus is easily spread by mechanical inoculation (Brunt, 1966) and 

can be spread by flailing (Mowat, 1987). Only the ringspot viruses are seed-borne. 

Most narcissus stocks are heavily infested with viruses, and while their effects may be 

generally less dramatic than those of stem nematode or base rot, they are important 

and their significance should not be under-estimated by growers. 

Narcissus crops may be affected by a number of physiological disorders, including 

damage from frost, waterlogging and sun scorch. Damage due to chemical or 

other treatment can be due to herbicides, HWT, formaldehyde or mechanical 

damage. ‘Grassiness’ (‘horses teeth’) can result when the main shoot of a bulb is

62 G.R. Hanks 

lost, resulting in the production of many offsets. The disorders ‘chocolate spot’ 

and ‘root rot’ may have the appearance of fungal diseases, but no pathogen has 

been associated with them. Finally, there are a number of disorders in flower development, 

resulting in the death or deformity of the flower bud or floral organs. 

STANDARD PRODUCTION OF NARCISSUS BULBS IN THE FIELD 

Planting material 

In obtaining bulbs as planting material, the potential grower needs to consider 

cultivar, source and quality, and bulb grade. 

Cultivars 

Although many narcissus cultivars are grown, few are cultivated in significant 

amounts, so stocks of relatively few cultivars can be obtained in quantity. The choice 

of planting stock will depend on the characteristics required (e.g., flower quality 

or galanthamine concentration) and the amount of biomass produced, linked with 

the price of different cultivars; these aspects are considered in Chapter 10 of this 

volume. General sources of information on the main commercial cultivars grown 

include ADAS (1982b, 1985a) and IFC (undated). The classified list of narcissus 

names is published by the Royal Horticultural Society (RHS), the International 

Registration Authority for the genus (Kington, 1998). The American Daffodil Society 

(ADS) publishes an abridged list (Throckmorton, 1989) and an illustrated database 

on CD (ADS, in press). Data from cultivar trials have been reported from the 

UK (Fry and Shepherd, 1961; ADAS, 1963, 1967, 1971; Hanks, 1994a; Hanks and 

Withers, 1998), Czechoslovakia (Petrová, 1983), Germany (Loeser, 1979), Poland 

(Szlachetka, 1989) and the USA (Nelson, 1988). Dutch information on narcissus 

cultivars appears in the annual guide Narcissengids, published by the Coöperatieve 

Nederlandse Bloembollencentraal, Lisse, and in the descriptive lists of ornamental plant 

cultivars issued by RIVRO (the National Institute of Crop Variety Research). 

Further information appears in articles in the periodicals Bloembollencultuur, 

HOBAHO and Vakblad voor de Bloemisterij. There are several public collections of narcissus 

cultivars in the Netherlands, including the CNB-Showtuin ‘Hein Schrama’ 

(Heemstede), Narcissen Showtuin K.J. van der Veek (Burgervlotbrug), HOBAHO 

Keurtuin ‘De Buitenhof’ (Lisse) and ‘Hortus Bulborum’, a collection of old cultivars 

(Limmen) (P.J.M. Vreeburg, personal communication). In the UK, bulb cultivars 

are displayed at Springfields Garden (Spalding) and are trialled at the RHS garden 

at Wisley, while there are a number of private collections under the aegis of the 

National Council for the Conservation of Plants and Gardens (NCCPG, 1999). 

Bulb source and quality 

Bulbs may be obtained by private sales or from auctions, or may be grown under 

contract arrangements. Obviously, a reputable source should be used to ensure 

trueness-to-type and quality, and, if possible, the stock should be inspected during 

the previous growing season. Stocks should be checked for the main pests and


diseases – stem nematode, bulb rots, foliar diseases and viruses. Bulb stocks should 

be obtained in good time to carry out HWT prior to prompt planting, and bulbs 

should be properly stored until planting. 

In the EU, a ‘Plant Passport’ scheme is in operation for bulb sales within the 

‘Single Market’. In the UK, for example, bulb stocks are entered by the grower 

with the Plant Health and Seeds Inspectorate (PHSI) of MAFF. Stocks are then 

inspected for stem nematode, the only EU quarantine pest of narcissus, in a growing 

season inspection (GSI), and a dry bulb inspection (DBI) is carried out after 

cleaning and grading bulbs to ensure freedom from soil. Bulbs are eligible for the 

grower to issue a plant passport if both inspections are passed; if they are not, it 

may be permitted for the bulbs to be sold for retail sales or landscape use only. 

Similar arrangements exist in other EU countries. For exporting bulbs to ‘third 

countries’ (i.e., outside the EU), plant health inspections and their criteria depend 

on the requirements of the importing country: for example, for export to the USA 

and Canada, the requirements also include strict freedom from soil on the bulbs, 

that bulbs are grown on certified PCN-free land, and that they undergo field 

inspections for freedom from other pests and diseases as well as pre-export inspections. 

When narcissus bulbs are bought in from a different climate, they may take 

some time to become synchronised with local stocks. Abbiss and Craze (1948) 

showed that Dutch stocks took at least three years to synchronise with local stocks 

in south-west England. 

Most ordinary narcissus stocks have widespread virus infection (Stone, 1973; 

Brunt, 1980), and VT propagation schemes offer the best opportunity for improving 

stocks (Hollings and Stone, 1979). Limited quantities of ‘virus-free’ narcissus 

stocks, more properly called virus-tested (VT) or virus-indexed stocks, are available 

from nuclear stocks produced from meristem-tip culture or from indexed plants 

(Brunt, 1985; Asjes, 1990; Lawson, 1990). VT stocks of narcissus should be grown 

in 50 m isolation from ordinary stocks (Broadbent et al., 1962) or in sterilised substrate 

in vector-proof fine mesh tunnels (Moore et al., 1979). VT stocks offer the 

general advantages of improved vigour. VT bulbs may have lower critical weights 

for flowering, and more, larger and brighter flowers (Stone, 1973). VT stocks may 

show increased bulb yields of 10–20%, compared with ordinary stocks, as well as 

earlier growth (Sutton et al., 1986, 1988). In some trials there was a consistent yield 

advantage for VT stocks over ordinary stocks in Cornwall, but not in Lincolnshire, 

suggesting that VT stocks were able to take advantage of the different conditions in 

Cornwall (Hanks, 1992a). Much greater advantages of VT stocks were reported in 

New Zealand by Allen and McIntosh (1994). When growing VT stocks, cultural 

practices may have to be adjusted to take account of their greater vigour, e.g., using 

lower planting densities. VT stocks of some cultivars are re-infected only slowly by 

viruses if grown in isolation (Hanks, 1997). 

In the UK, there are three grades of VT certification for narcissus under the 

voluntary Plant Health Propagation Scheme: (1) Virus-Tested Mother Stocks are 

the highest grade, strictly maintained in vector-proof conditions with zero tolerance 

for pests, diseases and off-types; (2) Foundation, field-grown stocks grown 

in isolation with low tolerance for pests and diseases (e.g., 0.05% for severe virus 

symptoms and 0.5% for mild virus symptoms); and (3) Élite, for which isolation 

and tolerances are less exacting (e.g., 2% for severe virus symptoms) (MAFF 

Plant Health Division, 1999). Methods for selecting, producing and certifying

64 G.R. Hanks 

pathogen-tested nuclear stock of narcissus have been described (Anon., 1993). 

Commercial ELISA testing services for narcissus are becoming available (e.g., 

Monro and Johnstone, 1992). However, ‘virus-freedom’ may be difficult to define, 

given the advances in virus identification and detection (e.g., Mowat et al., 1988a,b, 

1989; Boonekamp et al., 1990; Langeveld et al., 1997; Sochacki et al., 1997). 

Narcissus bulbs from glasshouse forcing are commonly reclaimed and can be 

‘plant passported’ for sale, but such ‘ex-forced’ bulbs must be labelled as such. 

Although they may be somewhat small and desiccated, if properly hot-water 

treated and grown-on for two or three years, good growth should be obtained for 

an economical starting price. 

Bulb size and shape 

Yields are mainly controlled by the grade of bulbs and their planting density. 

Narcissus bulbs are graded by circumference, with bulbs of the middle sizes 

(12–16 cm) being in demand for dry bulb sales and bulb forcing. Small bulbs 

(10–12 cm grade) are considered good planting material, since their percentage 

weight increase can be high. Large bulbs (>16 cm) are often re-planted in a stock 

because they are said to ‘increase vigour’, and they have been shown to emerge 

earlier and have larger flowers than small bulbs (Strojny, 1975). The presence of 

many small, round bulbs in a stock may indicate declining vigour, and it has been 

suggested that bulbs


although the growing season is delayed, growth then takes place under higher 

light levels and longer days. Rees (1972) compared the growth of identical stocks 

of narcissus ‘Fortune’ grown in south-west and eastern England. In the southwest, 

emergence, anthesis and senescence occurred 2–4 weeks earlier than in the 

east. After planting, the rate of dry weight loss was faster in the south-west and 

minimum plant weight was reached sooner, presumably because of the higher soil 

temperatures. Although bulb weight also increased sooner in the south-west, the 

eventual bulb yields were higher in the east. Annual temperatures and rainfall for 

important bulb-growing areas are given in Figure 4.1. 

There are large differences between years in narcissus yield, quality and timing, 

much of which may be attributable to weather conditions (Hanks, 1996a). 

Szlachetka and Romanowska (1990) carried out long-term trials, and reported 

that weather conditions, especially in December, had an effect on bulb yield 

greater than the effect of the grade of planting material used. Where the mean 

monthly air temperature was above 0 °C or slightly below (–0.8 °C) and the soil was 

not frozen, bulb yield was high; if the temperature was below –3.6 °C and the soil 

was frozen, yield was decreased by 14–48%. 

Narcissus bulbs are generally regarded as suitable for naturalising in US 

climatic zones 4 (annual minimum temperature about –30 °C) to 8 (about –10 °C) 

(De Hertogh, 1989; De Hertogh et al., 1993). Information on the hardiness and 

performance of a range of cultivars is available from trials such as those described 

by Klingaman and Eaton (1983) and Nelson (1988). In the Netherlands and 

similar climates, narcissus crops are covered (e.g., with straw) to protect them from 

frost or other injury. In the UK, crop covers are not used, as soils seldom freeze to 

sufficient depths to damage bulbs of most cultivars. Tazetta narcissus cultivars are 

hardy in the Isles of Scilly, whereas cultivars with Tazetta parentage (e.g., ‘Têteà-Tête’) 

may be damaged by low temperatures in eastern England. Data are 

available on the cold tolerance of narcissus cultivars, although there are some discrepancies 

between studies, perhaps because of the different experimental techniques 

used (van der Valk, 1971; Sakai and Yoshie, 1984). 

An annual rainfall of about 100cm is considered ideal for narcissus growing. 

Evenly distributed rainfall favours crop growth, especially in April to June when 

the bulb is growing rapidly, and also facilitates bulb planting and lifting. In 

appropriate climates, the availability of irrigation water should be considered. 

Waterlogging reduces crop growth, although narcissus are not specially prone to 

damage and can tolerate waterlogging if the water is well aerated (Gibson, 1935). 

However, wet soils assist the spread of stem nematode in the soil water and in 

waterlogged furrows. Growing in light sandy soil with a controlled water table is 

the ideal bulb-growing environment, except that the spread of nematodes is favoured. 

In windy situations crops may be flattened, reducing yields. MacKerron and 

Waister (1975) investigated the effects of shelter on narcissus performance, and 

concluded that reducing the wind run from 164 to 118 km/day was probably not 

economic for increasing yields. 

Topography 

Large flat fields are the most economic to farm, while small, sloping fields present 

a number of practical problems (Figure 4.2). Warm sites, such as south-facing

Figure 4.1 Mean monthly weather data for Falmouth (south-west England), Kirton 

(eastern England), Den Helder (the Netherlands) and Mt Vernon (Washington 

State). Data are long-term means, for at least 30 years to 1999, from 

Washington Post historical weather data and Horticulture Research 

International.


slopes, may favour the development of bulb rots, while nearby shelter (hedges, 

etc.) may favour the survival of large narcissus fly. Nearby bulbs in gardens, field 

margins, etc., may also act as a source of infection of large narcissus fly. Sloping 

sites can potentially give rise to problems with surface run-off of pesticides. 

At the present time, pesticide applications are an essential part of narcissus 

husbandry, and topography and soil type may have an effect on pesticide run-off. 

Field operations associated with growing narcissus, potatoes, etc., often result in 

the movement of turbid water, and soil-rich run-off from ill timed operations 

Figure 4.2 Typical narcissus fields of south Lincolnshire (top) and old daffodil beds 

on St. Marys, Isles of Scilly (bottom) (Photographs: Horticulture Research 

International and Andrew Tompsett, respectively).

68 G.R. Hanks 

(as well as natural erosion) on sloping sites can move pesticide residues into watercourses. 

Concentration of pesticides has been recorded in colluvium and in the 

sediment of surface pools where the watertable was high (Harrod et al., 1991; 

Harrod and Rickson, 1994). 

Soil 

Silts or very fine sandy loams are ideal for narcissus growing. In the UK, bulbs are 

grown successfully on a variety of silt, silt loam, brick-earth and peat soils. In the 

Netherlands, narcissus were traditionally grown in sandy soil and it was once 

considered that bulb quality could not be maintained on heavy silt clay soils 

(de Vroomen and de Groot, 1991). However, they are now grown on the heavier 

polders, factors involved including the pressure on prime agricultural land and 

the need, on sandy soils with a high water content, to practice regular soil sterilisation 

to control nematodes. There have been few comparisons of growth in different 

soil types, although Szlachetka (1976) reported better narcissus growth in light 

black-earth soils than in heavy alluvial soils. It has been reported that narcissus 

bulbs were larger and more susceptible to base rot when grown on peat, rather 

than mineral, soils (ADAS, 1982a). 

According to ADAS (1985a), bulb soils should be deep, fertile, well drained 

and moisture retentive (water holding capacity >40 mm per 300 mm depth), 

while the level of organic matter should be >3% and the pH value should generally 

be between 6.0 and 7.5. In the bulb growing areas of eastern England, 

typical mineral soils have 2.5–3.0% organic matter, while peat soils may have up 

to 25% (J.B. Briggs, personal communication). Weed control is difficult in highly 

organic soils. Narcissus bulbs are susceptible to poor growth due to compaction 

(De Haan and van der Valk, 1971). Although the observed root spread of narcissus 

was similar in different soil types, it was low in comparison with tulips, so a 

good soil structure is needed (Rees, 1972). Heavy soils, stones and clods can 

damage bulbs and make lifting difficult, requiring the use of stone and clod 

removers. In ‘sticky’ soils it may be difficult to produce soil-free bulbs unless 

bulbs are washed, a process which spreads diseases and creates disposal problems 

(ADAS, 1987). 

Crop rotation and site history 

Crop rotation should aim to maintain a good soil structure and reduce pest, 

disease and weed problems. Leys, peas, barley or early potatoes often form part 

of the rotation with narcissus. Preceding crops leaving a high residue (e.g., cauliflowers) 

should be avoided, as high levels of soil nitrogen may encourage basal rot 

(Hanks et al., 1998a). Rotations should allow adequate time for soil preparation 

before bulb planting. Narcissus should not follow crops treated with sulphonylurea 

herbicides, which can cause morphological damage (Greenfield, 1987). With 

‘two-year-down’ growing, foliar fungal diseases generally become a problem in the 

second year, so first- and second-year narcissus crops should not be planted adjacent 

to each other (Melville, 1980). 

At least four years should elapse between growing narcissus in a rotation, or 

six years where stem nematode has been found. In sandy soils, deep ploughing


can be used to bring up fresh soil and allow a shorter rotation. Where tulips are 

grown in the same rotation, they should follow narcissus, since the tulip race of 

stem nematode attacks narcissus bulbs but the narcissus race does not attack 

tulips. Other hosts of the stem nematode, which should be avoided, include (1) 

crops such as onion, maize, peas, beans, sugar beet, mangold, carrot, turnip, red 

clover and strawberries, (2) bulbs such as bluebells and hyacinth, and (3) weeds 

such as speedwell, scarlet pimpernel, chickweed, cleavers and black bindweed 

(Jones and Jones, 1984). For the export of bulbs to certain countries, they must 

be grown on soil certified free of potato cyst nematode (PCN): although PCN 

does not attack narcissus, the cysts can be carried on soil attached to dry bulbs. 

Spores of the base rot fungus are widespread, and have been found even in soils 

not previously known to have grown narcissus (Price, 1975a,b). Base rot spores 

may remain viable for up to 10 years, at least under artificial conditions (C.A. 

Linfield, personal communication). Diagnostic methods for the base rot fungus 

have been investigated (Linfield, 1993). Field populations of Fusarium oxysporum 

f.sp. narcissi are being characterised using molecular tools with the ultimate goal 

of developing sensitive and specific detection systems for this pathogen in bulbs 

and soils. These in turn may lead to diagnostics capable of predicting the disease 

potential of fields selected for narcissus production (J.H. Carder, personal 

communication). 

Bulbs left behind in the ground after harvesting (‘volunteers’ or ‘groundkeepers’) 

are difficult to eliminate, but the removal of these bulbs is important in controlling 

pests and diseases. Contact herbicides (such as paraquat or glyphosate), 

cultivation and picking by hand should be used (Tompsett, 1974). 

Economic factors 

The site chosen for narcissus bulb production should take account of the local 

availability of experienced labour, if required for labour-intensive operations such 

as flower cropping or bulb cleaning. The location of markets, local infrastructure 

and availability of logistics services should also be considered. 

Pre-planting operations in the field 

Cultivation 

Whenever soil is compacted, or suspected of being so, it should be sub-soiled 

(deep-ploughed) prior to growing narcissus. Good agricultural practices must be 

used, for example, carrying out sub-soiling when the soil is dry enough to burst 

the compacted layer but not wet enough to cause smearing. Perennial weeds 

should be removed by cultivation or by using a translocated herbicide. The land 

should be ploughed well in advance of planting, and when the soil is dry enough 

for structure to be maintained. Either the whole field should be ploughed and 

roadways taken out later, or permanent headlands should be established. After 

ploughing, the land may need further cultivation, to 25 cm depth in clay or loam, 

or 35 cm in sand. If the bulbs are not ready to plant and conditions are dry, the 

prepared land may be rolled to conserve moisture. In appropriate soils, the use of 

stone and clod separators before planting is becoming more usual. To avoid soil

70 G.R. Hanks 

compaction and its effects on clod formation and reduced crop growth, bed 

formers are used to produce an even soil particle density prior to planting. All 

cultivations should take account of the need to reduce soil compaction by reducing 

machinery movements and the area affected by wheelings. 

Soil disinfection 

In the UK, where bulbs are grown on heavier soil, it is not usual to use soil disinfection 

on a field scale. However, nematodes move more freely in sandy soils, and in 

the Netherlands soil disinfection is routinely used in bulb growing. Prior to assigning 

a field for narcissus production, soil samples should be sent to a diagnostics 

laboratory for identification and counts of nematodes present, including stem 

nematode, virus vector nematodes and PCN, as appropriate. Sterilants used in the 

Netherlands include metam-sodium and dichloropropene, which should be used 

at sufficient depth to control the target organisms (up to 40 cm for Pratylenchus). 

Soil disinfection may also be used to control soil-borne fungi such as white root rot 

(Mantell and Wheeler, 1973). 

Fertilisers 

Fertilisation for narcissus should aim to maintain adequate levels of phosphate 

(P 2 O 5 ) and potash (K 2 O) in the soil and to supply adequate (but not excessive) 

nitrogen (N). Nutritional experiments with narcissus have proved difficult because 

of the reserves within the bulbs (Bould, 1939; Hewitt and Miles, 1954). Long-term 

experiments showed that there is little short-term response to N or P 2 O 5 , that 

adequate K 2 O is needed, and that fertilisers do affect growth in subsequent years 

(NAAS, 1961; Wallis 1966, 1967b, 1968). K is beneficial in the second year of 

growth (Fodor and Sólymos, 1975). 

Fertiliser recommendations for general bulb growing under UK conditions are 

given in MAFF (1994). Soil samples should be taken well in advance of planting 

for the determination of P 2 O 5 , K 2 O and magnesium (Mg) levels, while N requirements 

are usually based on the previous cropping of the land, the highest rates 

being applied following demanding crops such as cereals. The maximum rates 

suggested, where the index has fallen to zero for a particular nutrient, were (in kg/ha) 

100 N, 125 P 2 O 5 , 250 K 2 O and 150 Mg, after deducting the amount of nutrients 

applied as organic manures. If necessary to apply magnesium, kieserite or calcined 

magnesite is used, unless liming is also necessary, in which case magnesian 

limestone is used. P 2 O 5 , K 2 O and Mg are usually applied after ploughing and 

worked in before planting, but it is recommended that N is applied later as a topdressing, 

shortly before crop emergence (to avoid scorching the foliage); this 

reduces leaching by winter rainfall. In practice, growers may prefer to apply a 

compound fertiliser before planting. Manganese deficiency may occur in narcissus 

grown on soil where the pH is high, and manganese sprays should be applied in 

advance of deficiency becoming evident. 

There may be advantages to applying fertilisers as split-dose applications (preplanting 

and as a spring top-dressing) (Parker, 1935) and to counter the effect of 

rain (Lyakh, 1988), but there was no advantage of using repeated high rates of N 

(Lees, 1960). No advantage was demonstrated in trials of applying fertiliser into


the ridges at planting, instead of broadcast (ADAS, 1970). No fertilisers are 

applied in the second year of the crop, unless growth is obviously poor in the first, 

in which case a low rate of N (50 kg/ha) is top-dressed before crop emergence. 

Chilvers and Daft (1980, 1981) reported on the widespread occurrence of endomycorrhizal 

fungi (Glomus spp.) in narcissus in Scotland. Mycorrhizal infections 

assist in the uptake of phosphates and possibly other nutrients, and attempts had 

been made to introduce more active mycorrhizal species into field situations. Iqbal 

and Firdaus (1986a,b) described mycorrhizal infections in N. poeticus. In pot experiments 

with N. tazetta, inoculation with Azospirillum spp. improved productivity in 

sandy soils even when nitrogen fertilisers were applied (Naggar and Mahmoud, 

1994). 

From hydroponics experiments, Ruamrungsri et al. (1996a,b, 1997) reported 

decreased bulb dry weight only where nitrogen (N), calcium (Ca) or magnesium 

(Mg) were omitted from the nutrient solution, although flower quality was not 

affected. Omitting N resulted in stunted shoot growth, with small, thin yellow 

leaves, while omitting Ca depressed root and shoot growth. Omitting Mg gave 

severe interveinal chlorosis near the leaf tips, omitting iron (Fe) gave chlorosis 

near the base of leaves, and omitting boron (B) resulted in water-soaked areas in 

the basal part of leaves. The only other report concerning trace element effects 

was from a study that found few differences in yields between different fertiliser 

treatments, but that flower yield was increased when B was applied (Emsweller 

et al., 1938). The importance of Ca for yields was also shown by Hewitt and Miles 

(1954). Sun et al. (1991) reported that bulb treatment with paclobutrazol increased 

Fe uptake and reduced zinc (Zn) uptake in hydroponically grown plants, but the 

effects on growth were not reported. 

Dutch investigations indicated that excessive amounts of fertiliser are often used 

on bulb crops, and that narcissus have lower N requirements than other bulb 

crops (van Berkum, 1987; van Berkum and Braam, 1991). A level of 125 kg N/ha 

was recommended, and it was suggested that soil mineral nitrogen should be analysed 

and the amount of N applied should be reduced by this amount. In the UK, 

a scale of N applications has been recommended, depending on soil type, from a 

maximum of 120 kg/ha in sandy or shallow soils, decreasing to zero in peaty soils 

(C.R. Rahn, personal communication). Excessive N levels are reported to lead to 

more bulb rots, splitting and bruising in narcissus (Biekart, 1930; McClellan and 

Stuart, 1947; Rikhter, 1976; Hanks et al., 1998a); for example, Rikhter (1976) 

found highest yields with N applied at 120 kg/ha, with more fungal disease at 

180 kg/ha. The lowest rates of base rot occurred when potassium was added 

(McClellan and Stuart, 1947). The excessive use of phosphate fertiliser remains an 

important consideration in Dutch bulb growing (Pasterkamp et al., 1999). 

Organic fertilisers may be used, but high rates should not be applied shortly 

before planting. Allen (1938) reported that ammonium sulphate (0.5 t/ha), 5:10:5 

(N:P:K) fertiliser (1.5 t/ha) and rotted manure (4 t/ha) each about doubled bulb 

yield. Well rotted farm-yard manure applied to the previous crop at up to 75 t/ha 

may be useful in soils where organic matter is low, but trials showed no advantages 

of using particular types of bulky organic fertilisers (Lees, 1961). The use of sewage 

sludge on bulbs is not well known, but its use is not recommended where bulbs are 

being grown for overseas sales because of the possibility of contamination with 

PCN. Properly composted waste from bulb growing (including waste containing

72 G.R. Hanks 

bulbs with base rot) can be safely spread on bulb fields, according to Dutch trials 

(van Dijk, 1990). Narcissus growing produces 3–5m3 /ha of waste from the field or 

1 t/ha from bulb forcing (Bouma, 1990). Care should be exercised in applying 

plant wastes from processing to cropping land, because of the possible effects of 

residual solvents present. 

Pre-planting bulb treatments 

Following the receipt of bulbs, the main treatments to be considered before planting 

are storage and hot-water treatment (HWT). In all bulb handling operations, a 

reasonable standard of hygiene should be maintained, particularly of containers, 

bulb stores and equipment, as soil and other debris can be a major source of stem 

nematode, bulb scale mite, and fungal propagules. Nematode ‘wool’ (the dehydrated 

fourth stage juvenile stage of the stem nematode) can survive for 25 years 

in its dry state. For use at room temperature, phenols (including cresylic acid) and 

iodine/phosphoric acid disinfectants were most effective in killing stem nematode 

(Lole, 1990). Alternatively, bins, etc., can be treated in HWT tanks for 10 minutes 

at 50 °C. Stored bulbs can be fumigated by specialist contractors with methyl 

bromide to control bulb scale mite, without phytotoxicity (Gurney and Gandy, 

1974; Murdoch, 1975; Powell, 1977). 

Bulb storage – general 

Ideally, bulbs should be stored after receipt at 17–18 °C in a controlled-temperature 

store with good air movement, some exchange of fresh air, and a relative humidity 

below 75%. Temperatures below 17 °C slow shoot development, and high temperatures 

will encourage base rot. In practice, bulbs are often stored outdoors (in 

which case they should be protected from sun and rain) or in sheds at about ambient 

temperatures. If stored in bulb trays, air circulation may be adequate; if stored 

in loose bulk or in bulk bins, it will be necessary to maintain some air movement 

through the bulbs, through ducts or fans, to prevent dampness which could lead 

to premature rooting and fungal growth. Roots of narcissus bulbs form over a 

short time and emerge simultaneously, so if they grow in storage and are broken 

at planting, this damage can be serious (Rees, 1972). When narcissus roots were 

repeatedly excised (up to four times), new roots developed from the base plate, 

but their number was progressively reduced and, increasingly, plants failed to 

flower (Yasuda and Fuji, 1963). 

Hot-water treatment (HWT) 

HWT is the key aspect of growing narcissus bulbs, and its practice is described in 

Gratwick and Southey (1986) and ADAS (1985b). It is essential to apply HWT 

(colloquially called ‘sterilising’ in the UK) to all narcissus planting stocks to control 

stem nematode, and other pests are also killed by HWT. It is important to use 

HWT even if bulb stocks have no history of stem nematode, as very few nematodes 

are needed in a bulb to cause its destruction (Hesling, 1971), while the treatment 

of stem nematodes at sub-lethal temperatures can result in only temporary inactivation 

(Hastings et al., 1952). Early experiments on the HWT of narcissus were


carried out by Hewitt (1914), but the treatments used (1–6 hours at 48.9 °C) 

resulted in the death of the bulbs. The technique was further developed by Ramsbottom 

(1918, 1919), Staniland (1933) and Staniland and Barber (1937), resulting 

in a standard recommendation of 3 hours at 43.5 °C. The early studies have been 

fully described by several authors (Slootweg, 1962; Turquand, 1966; Tompsett, 

1982; Lane, 1984; Gratwick and Southey, 1986). Treatments were later increased 

to 3 hours at 44.5 °C or 4 hours at 43.5 °C, to improve control (e.g., van Slogteren, 

1931; Chitwood and Blanton, 1941; Woodville and Morgan, 1961). Lees (1963) 

gave the critical treatments for crop damage. Growth was not impaired by extending 

a 3 hour treatment to 46.1 °C, nor by extending a 43.5 °C treatment to 6½ 

hours, but a 3 hour treatment at 47.2 °C impaired vigor and at 54.4 °C a 45 minute 

treatment was lethal. For N. tazetta var. chinensis, Lin et al. (1987) reported that 

bulbs grew normally after HWT for 45 minutes at 50 °C or 25 minutes at 55 °C, 

although complete control of the target nematodes (in this case Aphelenchoides spp.) 

was not achieved. The chemicals applied do not penetrate beneath the bulb scales 

at ambient temperatures unless a vacuum treatment is applied (Newton et al., 

1933). ‘Vapour heat treatment’ was also investigated, but the regime necessary to 

control stem nematodes – 8 hours at 47.7 °C – had adverse effects on the crop 

(Chitwood and Blanton, 1941). Other methods of treating stem nematode (such as 

nematicidal dips or field applications) are either ineffective, or suitable chemicals 

are not now available (Hesling, 1971; Damadzadeh and Hague, 1979; Windrich, 

1986). 

With two-year-down growing, effective HWT becomes even more important, 

and this is reflected in the UK recommendation to treat all planting stocks for a 

full 3-hour period at a temperature of 44.4 °C. The treatment period is usually 

taken as starting when the tank temperature regains 44.4 °C after the cooling effect 

of loading the bulbs. In the Netherlands, using one-year-down growing and particularly 

where soil sterilisation is used to control nematodes, it is usual to apply 

a milder treatment (2 hours at 43 °C), adequate to control mites and narcissus fly 

larvae. Where a nematode problem is suspected, or when using two-year-down 

growing, a longer, hotter regime (4 hours at 47 °C) is recommended, and a 4 hour 

treatment at 48 °C is used for ex-forced bulbs (Vreeburg et al., 1999). At the time 

of writing, anecdotal evidence is that growers in the UK and the Netherlands are 

treating bulbs at up to 49 °C. Warm-storage and pre-soaking are necessary prior to 

HWT in these cases (see below). 

A variety of designs of HWT tanks is used, the usual type being a front-loaded 

design where bulk bins of bulbs are loaded and unloaded using a fork-lift truck 

whilst the dip solution is temporarily pumped to a holding (or ‘slave’) tank (Figure 

4.3). Tank capacity varies depending on the scale of the operation, but may be 

from 0.5 to 10 t of bulbs. Since the temperature of HWT is critical for killing nematodes 

without causing unnecessary damage to the bulbs, good temperature control 

and tank circulation are key factors in HWT tank design. Many other factors 

are also important, such as rapid heating, good insulation, a suitable water:bulb 

ratio, temperature monitoring and recording, operator safety and convenience of 

use. Aspects of the design and use of HWT tanks have been described by Gratwick 

and Southey (1986). Although HWT may be used to control pests and diseases in a 

variety of other plant material, flower bulbs (in particular narcissus) are the main 

subjects for HWT, and an HWT facility is likely to be the major dedicated outlay

74 G.R. Hanks 

Figure 4.3 Modern front-loading hot-water treatment tanks. Two four-tonne units 

with overhead slave tank for holding dip when loading and unloading 

bulbs (Photograph: Lyn Secker, Secker Welding, Holbeach). 

for bulb producers. HWT tanks can also be utilised for sterilising bulb containers 

and other equipment, and other produce requiring dipping in pesticide may 

make use of them at ambient temperatures. HWT of narcissus bulbs may be carried 

out under contract, but in UK it is usual for bulb growers to treat their own bulbs. 

The correct timing of HWT involves the overall logistics of the farming operation. 

To minimize crop damage, narcissus bulbs are treated after all flower initials 

have been formed and are visible under a hand-lens on dissection of the bud, but 

before the root initials have developed too far (shown by the roots erupting from 

the base plate). The internal stage of development (ISD) of ‘complete flower differentiation’, 

called ‘Stage Pc’ by bulb growers from the paracorolla (trumpet or 

cup), the last formed part of the flower bud, is illustrated in Preece and Morrison 

(1963), Cremer et al. (1974) and ADAS (1990b). In practice, this means that in the 

UK, for example, bulbs should be treated from late-July onwards, aiming to complete 

HWT before the end of August, a window of some four weeks. HWT tank 

capacity should be calculated to suit this treatment window and the cultivars grown. 

Narcissus cultivars which produce fine or early roots (Poeticus, Cyclamineus and 

Jonquilla cultivars) should be given HWT first (Benczur, 1976), while for mainstream 

cultivars the accepted order is small-cup, large-cup, then trumpet cultivars, 

although earlier HWT will give better control of base rot (ADAS, 1974; Millar, 

1976; Price and Briggs, 1976). There should be a gap of at least a week between 

removal of offsets and HWT, as recent offset removal can increase infection, especially 

in certain cultivars (Kruyer, 1978). Bulbs re-claimed from forcing may be 

given HWT early, in May or June (Vreeburg and Korsuize, 1991). Where flower 

quality is unimportant, an earlier start to HWT may be acceptable, although the 

critical development stages or earliest dates for avoiding damage to leaf initials has


not been reported; late HWT is probably more damaging, because of the potential 

loss of leaf area and roots. In cases where a problem with nematodes is suspected 

or known in a stock, bulbs should be lifted early (June), cleaned, graded and given 

HWT promptly. Earlier recommendations (e.g., Hastings and Newton, 1934) and 

some Dutch recommendations state that HWT in this case should be done within 

three to four weeks of lifting, but current recommendations in the UK are to treat 

sooner than this (J.B. Briggs, personal communication). The increase in stem nematode 

numbers in bulbs during storage was shown by Winfield and Hesling (1966). 

Even when applied correctly, HWT can reduce crop vigor, but this disadvantage 

is outweighed by the control of stem nematode and, in any case, the loss of vigour 

is insignificant in two-year growing. If HWT is carried out too early or too late, for 

too long a time or at too high a temperature, or following the storage of bulbs at 

low temperatures, a variety of damage occurs. This varies from mild damage to the 

flowers (unimportant where crops are being grown for processing), through damage 

to the leaves (from mottling of the leaf tips to severe distortion or stunting 

resulting in bulb yield loss) or roots (resulting in severe yield loss), to the death of 

the bulbs. Excessive HWT temperatures may result in the abnormal production of 

additional bulblets (Edwards, 1965; H.Y. Alkema, personal communication). 

Hot-water treatment – the use of chemicals 

While stem nematodes in the bulbs are killed by the high temperatures of HWT 

alone, chemicals are added to HWT tanks to control pests and diseases better. The 

basic material added is the disinfectant formaldehyde, used as ‘commercial formalin’ 

(containing 38–40% formaldehyde) (Hawker, 1944). For California, Qiu et al. 

(1993) reported that the standard HWT regime was 4 hours at 44 °C, but showed 

that such a treatment was sufficient to control stem nematode without added 

formaldehyde; using formaldehyde, 150 minutes was sufficient and caused no 

crop damage. However, the time for the centres of bulbs to attain the target temperature 

after placing in hot water should be added to these basic times: this was 

calculated as equal (in minutes) to –15 + 3.4x, where x is bulb circumference in 

cm (Qiu et al., 1993). Higher temperatures could be used for shorter times: without 

formaldehyde, stem nematode was controlled by 60 or 15 minute treatments 

at 46 or 48 °C, respectively, without crop damage; with formaldehyde, control was 

achieved by 90, 45 and 30 minute treatments at 46, 48 and 50 °C, respectively, but 

this caused crop damage. HWT for 4 hours at 44 °C reduced the number of fungal 

colonies (Penicillium sp., Fusarium oxysporum f.sp. narcissi and Mucor plumbeus) 

recovered from bulbs, but the effects of formaldehyde, glutaraldehyde and 

sodium hypochlorite in controlling these fungi were variable. 

Despite these possibilities for dispensing with a disinfectant, it is generally 

considered essential to add formaldehyde, which is effective at HWT temperatures 

in killing free-swimming stem nematodes (nematodes that escape from the bulbs 

into the tank dip and which are more resilient to high temperatures), as well as the 

spores of the base rot fungus. Formaldehyde is usually added as 5 litres commercial 

formalin per 1000 litres (0.2% a.i.). Increasing the concentration of formaldehyde 

gives little extra benefit in fungicidal activity, but can result in crop toxicity, 

with fewer and deformed flowers (Price and Briggs, 1976; Linfield, 1991), so any 

recommendations to use higher rates (e.g., Higgins, 1999) should be treated with

76 G.R. Hanks 

caution. Higher rates of formalin can also damage the base plates of bulbs, producing 

corky areas, especially when HWT is being carried out early or soon after 

lifting (Briggs, 1988), and in dwarf cultivars and others which form early, fine 

roots and which are susceptible to formaldehyde damage (Vreeburg, 1984b; van 

der Weijden, 1989). However, formaldehyde continues to be the material of 

choice for most growers because of its effectiveness, cheapness and availability. 

The use of formaldehyde is now coming under scrutiny due to health issues (Zell, 

1984), and it may be unavailable in some localities. Linfield (1991) evaluated 

alternative disinfectants for killing the chlamydospores of the base rot fungus 

under HWT conditions: commercial preparations of glutaraldehyde, hydrogen 

peroxide–peracetic acid and thiabendazole were highly effective and non-phytotoxic. 

A disinfectant based on peroxyacetic acid (peracetic acid) was effective in killing 

stem nematodes (both free-swimming and in the ‘wool’ stage) and chlamydospores 

of the base rot pathogen at HWT temperatures (Hanks and Linfield, 1997, 1999). 

In this study, peroxyacetic acid killed the chlamydospores of the base rot fungus 

within 1 hour, whereas total kill was not achieved with formaldehyde even after 4 

hours. Used in HWT, the peroxyacetic acid-based disinfectant was not phytotoxic 

to bulbs. Glutaraldehyde was shown to be effective against stem nematodes at HWT 

temperatures (M.J. Lole, personal communication). Alternatives such as bleach 

and chlorine dioxide are being evaluated by Chastagner (1999). 

Fungicides and insecticides may be added to HWT tanks along with formaldehyde. 

In the UK, thiabendazole is the material of choice for controlling the base rot 

fungus and other pathogens such as Penicillium. Thiabendazole, although used in 

agriculture and horticulture as a fungicide, was originally introduced to farming as 

an anthelminthic, and it appears to have useful activity against stem nematode in 

HWT (Hanks and Linfield, 1999). Because thiabendazole is more soluble at very 

acidic pH values, and, like many pesticides, is broken down by alkaline hydrolysis, 

for bulb dips it is used in an acidic formulation (‘Storite Clear Liquid’ in the UK). 

To further reduce the pH value of the dip an acidifier, sodium hydrogen sulphate 

(sodium bisulphate), is sometimes added to HWT tanks by growers on the basis of 

anecdotal evidence, and this is now being investigated experimentally (Hanks, 

1999). For stocks severely affected by base rot, thiabendazole is used in HWT even 

when it has been applied post-lifting, this double treatment providing the greatest 

initial reduction in the number of diseased bulbs (Hanks, 1996b). Prochloraz-based 

fungicides are also used, while a number of other fungicides have been found to 

be effective, including thiram, benomyl and carbendazim (ADAS, 1974, 1976; 

de Rooy, 1975). Fungicides used in the Netherlands include zineb/maneb, captan, 

benomyl, carbendazim and Topsin M, sometimes used in a mixture, especially for 

dwarf cultivars (Anon., 1987). Recent recommendations are for a combination of 

formaldehyde, captan, prochloraz and carbendazim (van der Weijden, 2000). Lower 

concentrations are used for ex-forced bulbs, because of the extra uptake by the 

drier bulbs. 

The insecticide chlorpyrifos may be added to HWT tanks to prevent subsequent 

infection of bulbs by the larvae of the large narcissus fly, although this gives 

protection only in the first year (Tones and Tompsett, 1990; Hanks and Linfield, 

1997). Cold dips or application at planting were less effective. 

It is usual to add a non-ionic wetter to HWT tanks to enhance the effects of 

pesticides, and other types of wetters should be avoided because of the possibility


of phytotoxicity (Wallis, 1966, 1967a). In trials with carbendazim, the addition of 

compounds believed to increase fungicide uptake (dimethyl sulphoxide, indolylacetic 

acid and hydrochloric acid) showed no benefit to disease control (ADAS, 

1976). The generation of foam in HWT tanks, which could reduce pesticidal effectiveness, 

may be reduced by the addition of an anti-foam preparation, which has no 

adverse effect on the crop (Tompsett, 1977). Where problems in handling bulbs 

are likely due to the applied pesticides, an anti-dust ‘sticker’ may be added, as is 

practised in the Netherlands (Anon., 1987). 

Some pesticide labels state that bulb dip solutions should be freshly made up 

each time, but this is impractical. It is usual to top up bulb dip tanks and to re-use 

them for as long as practical, providing gross contamination with debris (bulb 

skins, soil, etc.) can be avoided. After dipping, tanks are topped up with water and 

the appropriate amounts of disinfectant, pesticide, etc., are added so that the 

top-up is given at the original strength. Dutch recommendations to top-up formaldehyde 

at twice the original strength have now been modified, and 0.75% commercial 

formalin is now recommended (van der Weijden, 2000). For pesticides, 

manufacturers’ recommendations should be consulted, although these are often 

not specific. The thiabendazole fungicide ‘Storite Clear Liquid’ is an exception, as 

definite top-up procedures are given. As active ingredients can be lost in use, it is 

advisable to have the concentration of formaldehyde and pesticides determined by 

specialist laboratories for sets of samples, at least until experience of their behaviour 

is obtained, and particularly as little published guidance exists on the stability 

of pesticides in bulb dips. Recent Dutch recommendations suggest using slightly 

higher top-up rates for prochloraz, but using other pesticides at the original 

strength (van der Weijden, 2000). 

Treatments to reduce damage due to HWT 

HWT damage can be reduced by warm-storing bulbs for 1 week at 30 °C before 

HWT, a procedure used in south-west England to reduce damage to flowers in the 

year after HWT (Wood, 1944; Slootweg, 1962; Tompsett, 1975). Warm-storage is 

always recommended when treating sensitive narcissus such as Poeticus cultivars, 

and is beneficial in cultivar ‘Tête-à-Tête’ (ADAS, 1988b). The exact treatment is 

not critical, 3 to 8 days at 30 to 35 °C having been used (Rees and Turquand, 1967; 

Turquand and Rees, 1968). Such warm-storage, however, by partly desiccating 

the bulbs, induces stem nematode to migrate to the outside of the bulbs, often 

near the base plate, where they form ‘nematode wool’ which can escape into the 

bulb dip during HWT. Warm-storage is, therefore, used in conjunction with presoaking 

bulbs (for 3 or 4 hours or, preferably overnight, at ambient temperatures 

with formaldehyde) immediately before HWT, in order to hydrate the ‘wool’. 

With this regime a higher HWT temperature, 46 or 47 °C, is necessary to kill stem 

nematodes. It is important not to omit pre-soaking when pre-warming is used. 

Warm-storage in itself may decrease crop vigor (Wallis, 1965, 1967a). Warm 

storage appears to induce dormancy in the shoot initials, making them less sensitive 

to damage by high temperatures, and thus extends the period over which 

HWT can be safely used (Wallis, 1967c). The lower limit of the warm-storage effect 

is about 18 °C. If bulbs are stored at 18 °C for 2 weeks before HWT (where storage 

may otherwise have been at slightly lower ambient temperatures), HWT damage

78 G.R. Hanks 

can also be prevented, but without the need to pre-soak bulbs or to use a treatment 

temperature higher than 44.4 °C. By this means (sometimes called ‘partial 

pre-warming’), the window for the safe HWT of narcissus bulbs in eastern England 

can be extended to late-September (Hanks, 1995a). An equivalent treatment of 

1 week at 20 °C is recommended in some Dutch advisory material. 

There is a difference in Dutch and UK advice on the warm-storage of earlylifted 

bulbs known to be infected with stem nematode. The former suggests a warmstorage 

of at least one week at 30 °C after lifting, while in the UK it is stated that 

pre-warming should be avoided because of the danger of inducing the formation 

of ‘wool’ if the bulbs are allowed to dry out. 

Bulb treatment after HWT 

The increasing practice in UK is to plant bulbs soon after HWT, which means they 

can be planted while still damp. This avoids the need for re-drying and storage 

and reduces handling, but it means that the bulbs are planted into relatively warm 

soil in August, possibly encouraging base rot. When removed from the HWT tank, 

the bulbs should be allowed to drain and then should be cooled, ventilated and 

surface dried using fans in a drying wall or similar arrangement. Although there is 

no evidence from trials to show a distinct benefit from rapid cooling to ambient 

temperatures after HWT (Tompsett, 1973), this seems sensible. From the viewpoint 

of operator safety, the ventilation of bulb handling areas is critical, especially 

when bulbs are being removed from the HWT tanks and moved to the drying 

facility. As an alternative to immediate planting, bulbs can be dried after HWT, 

followed by appropriate storage and replanting in September, when soil temperatures 

have fallen sufficiently to slow the development of base rot. In this case, 

rapid drying is important in controlling base rot (Hawker, 1935, 1940). 

Other pre-planting pesticide applications 

The application of insecticides, other than in HWT, has been evaluated for the 

control of large narcissus fly. Separate pre-planting dips of chlorpyrifos were less 

effective than using the material in HWT (Tones and Tompsett, 1990). One-hour 

dips, either immediately after lifting or before planting, were evaluated by Bogatko 

(1988) and Bogatko and Mynett (1990): several insecticides were found to be 

effective in controlling larvae (including isofenphos and carbofuran). 

Bulb storage treatments to manipulate growth 

In addition to the considerations relating to bulb storage generally, storage 

temperatures may be altered, in the period before planting, to manipulate the 

subsequent growth of the crop. This was investigated by Rasmussen (1976b), who 

compared the growth of narcissus bulbs after storage at ambient outdoor temperatures 

and planting bulbs in September, with storage at 20 or 23 °C and planting in 

October. Emergence, but not anthesis or senescence, was delayed by controlledtemperature 

storage and later planting. Better yields of bulbs or flowers resulted 

from using the higher temperatures, such that best results were obtained from the 

ambient regime in warm winters and from 23 °C storage in cool winters. More


usually, however, interest has centred on the cool storage of bulbs before planting 

as a means of advancing (or ‘forwarding’) field-grown cut-flowers. In south-west 

England, ‘pre-cooling’ narcissus bulbs for 2–6 weeks at 9 °C before planting gives 

flowers earlier and over a longer period, although the flowers may be of poorer 

quality and subsequent bulb yield is reduced (Rees and Wallis, 1970; Rees, 1972; 

ADAS, 1982d). The optimum treatment for advancing flowering was 6 weeks at 

9 °C, with no advantage of using longer or colder treatments or of augmenting the 

treatment by covering the growing crops with polythene film (ADAS, 1982c; Flint, 

1983). Flowering dates 8–28 days earlier than untreated controls were reported in 

these studies. Cocozza (1972) investigated pre-cooling before planting bulbs 

outside in Italy, successfully using a treatment at 3 °C for up to 6 weeks. In France, 

Le Nard (1975) reported that flowers were obtained 30–45 days earlier than from 

untreated bulbs, by warm storage (7 days at 34 °C) followed by cooling (8 weeks at 

9 °C) and covering rows with a narrow polythene tunnel. To produce late outdoor 

flowers, bulb storage at 20–26 °C from August to October has been investigated, 

but this delayed flowering by less than a week (ADAS, 1972). Tazetta narcissus 

can be warm-stored over winter for planting in spring (see under Production of 

Tazetta Narcissus). 

While most interest in these procedures relates to optimising crop growth and 

producing earlier flowers, they also offer the possibility of shifting the growth 

pattern to one more suited to particular needs, for example to fit a more convenient 

or programed production schedule. The effect of temperature on narcissus 

growth and development is being studied with a view to developing predictive 

models (Hanks et al., 1998b,c). As narcissus growth and yield show marked yearto-year 

differences (Rees, 1972; Hanks, 1996a), such crop models would be useful 

management tools. 

Bulb planting 

The main factor to consider is the type of growing system – whether to grow the 

bulbs in ridges or beds, and whether to lift bulbs annually (one-year-down growing) 

or every two years. Because it is considered more economic, a ‘two-year-down’ 

growing system in ridges has generally been adopted. Other factors to be considered 

include planting date, rate, depth and arrangement; the use of pesticides 

at planting, crop covers, cover crops and growing in nets also need to be considered. 

Planting and growing in beds or ridges 

Bulbs (including narcissus) were traditionally grown in flat beds and were planted 

and lifted annually by hand. Bulb planting and lifting are now mechanised. In the 

UK, bulb crops are now grown in ridges (as potatoes), and ridges are also used in 

the Netherlands where narcissus are grown on heavier soils. Growing bulbs in flat 

beds allows better utilisation of the available area, but large volumes of soil have 

to be moved in planting and lifting bulbs. Growing in ridges has been adopted 

largely on practical grounds, and is useful as it allows sharing of equipment with 

potato growing. 

Bulbs are planted in ridges using specialised planting machines that typically 

feed bulbs from a hopper, planting two rows at a time (Figure 4.4). The planting

80 G.R. Hanks 

Figure 4.4 A typical bulb planting machine feeding bulbs from a hopper into two 

ridges (Photograph: Horticulture Research International). 

rate should be calibrated by adjusting the hopper aperture or the speed of 

the delivery belt. High planting rates may be difficult to achieve with standard 

machinery, and blockages often occur, so that constant attention from the operator 

will be needed. When planting in flat beds, small or uniform bulbs are 

planted using a row planter, planting four rows at a time. Larger bulbs are planted 

using bed digging and filling machines that lift the soil and transfer it to the adjacent 

planting bed. 

‘One-’ or ‘two-year-down’ growing 

Rees et al. (1973) compared one-, two- and three-year-down narcissus growing in 

south-west England, and concluded that costs were too high for a one-year-down 

system to be economic. UK narcissus growers now use this two-year-down growing 

system, half of the stock being lifted and re-planted in alternate years. This 

‘biennial’ growing involves some loss of bulb yield, because an ideal planting 

density obviously cannot be achieved in both years. Too low a planting density is 

inefficient in the first year, while too high a density results in subsequent overcrowding. 

At a planting density of 10 t/ha, half the bulb weight increase occurred 

in each year in two-year growing, but at high densities all the increase was in the 

first year (Rees et al., 1973). Some data (ADAS, 1993) are illustrated in Figure 4.5. 

In ornamentals production, however, when most bulb growth takes place in the 

first year, this has the advantage that higher flower yields are obtained in year two, 

and there are high yields of relatively small bulbs which are advantageous for 

retail bulb sales because they produce more flowers per tonne. The flowers produced 

in the second year are unaffected by any adverse effects of HWT two years 

earlier, so quality is improved. There is also an element of ‘compensation’ in yield,

Disposable yield (t/ha) 

40 

30 

20 

10 

0 

20 25 30 35 15 20 25 30 

Planting density (t/ha) 

First 1-year cycle Second 1-year cycle 2-year cycle 


Figure 4.5 Narcissus bulb yields for one- and two-year-down growing (data from 

ADAS, 1993). 

if poor growth occurs in one of the two years of the growing cycle, for example 

due to early senescence or pesticide damage. Two-year-down growing has enabled 

UK narcissus growers to become very efficient by reducing the annual demand for 

labor, land, storage and other resources (ADAS, 1986b). In some cases, growers 

may leave narcissus crops down for a third year for economic reasons, perhaps 

because bulb prices are low or the flower crop is more important, but this is not 

a practice that is recommended as good husbandry. Other reports of trials 

comparing one- and two-year-down growing have been reported from Denmark 

(Rasmussen, 1976a) and Poland (Sochacki and Mynett, 1996), confirming the 

advantages of the two-year-down system. 

The main disadvantage of two-year-down growing is that HWT and other 

useful procedures (such as post-lifting fungicide treatment or high-temperature 

drying) can be carried out only in alternate years, so there is no opportunity every 

year to control stem nematode and other harmful organisms. Fungal pathogens in 

the soil and stem nematodes in bulbs can build to serious levels over a two-year 

period, while foliar fungal diseases (such as smoulder) are serious problems only 

in the second-year of narcissus crops. In the summer between the two growth 

periods, bulbs remain in the soil when temperatures are high and liable to encourage 

bulb rots, and the benefits of early bulb lifting (the avoidance of narcissus fly 

and aphid-borne virus infection late in the growing season) are lost. When 

narcissus are left in the ground, they begin growing earlier, and earlier rooting 

may allow more effective infection by soil-borne pathogens while soil temperatures 

are still high. Second-year narcissus shoot earlier, and may therefore be more 

damaged by frosts. Where base rot is a major problem, a temporary return to oneyear-down 

growing should be considered (Tompsett, 1984). 

When growing narcissus for processing, rather than as ornamentals, different 

considerations may apply. If high yields and the avoidance of pests and diseases are 

important, one-year-down growing may be more appropriate. Where the most economical 

growing system is required, two-year-down growing will be appropriate, 

but there is unlikely to be any advantage of growing cycles longer than two years.

82 G.R. Hanks 

Planting date 

The practice of planting narcissus bulbs soon after HWT has been mentioned 

above, and, since the date of HWT is dependent on the stage of the crop, this also 

governs the date of planting. Planting immediately after HWT, in August or early- 

September, means that the bulbs are planted into relatively warm soil, increasing 

the likelihood of infection by the base rot fungus when the roots are erupting from 

the base of the bulb (Gregory, 1932; Hawker, 1935). Dutch recommendations, and 

earlier recommendations for the UK, are to plant narcissus bulbs in (late-)September. 

It is usual to plant cultivars in the same order in which they received HWT. In 

one-year-down growing, there is a steady decline in bulb yield when bulbs are 

planted later than September. One study showed yields of 120% from September 

planting and only 63% from December planting (Wallace and Horton, 1935), 

although in another trial (Allen, 1938) significant yield loss did not occur until 

planting was even later. In two-year-down growing, however, poor growth in the 

first year would be compensated by better growth in the second. 

Planting density 

The rate of planting will depend on the vigor of the cultivar, bulb price, the 

percentage bulb weight increase and size of bulbs required, and the growing 

system adopted. In the UK, narcissus bulbs for two-year-down growing are often 

planted at densities between 12.5 and 17.5 t/ha. A planting density in the lower 

range would be used where vigorous bulbs were being planted, where a high rate 

of increase was required (for example, where bulking an expensive cultivar), or 

where large bulbs were required; a high density could be used, for example, when 

growing a cheap cultivar where the requirements were a reasonable rate of 

increase coupled with reducing land and labor requirements, or where a good 

yield of smaller grade bulbs was needed. Higher planting rates are more economic 

of land and labor: the preferred planting rate is the one which gives the best 

financial returns, rather than the greatest yield (Rees, 1972, 1975; ADAS, 1976). 

The effects of planting density and planting grade in ridges were investigated by 

Rees (1972) in eastern England. For smaller planting grades, the optimum densities 

were above the highest used in the trial. For larger grades, there were large differences 

in optimum densities and financial returns between years, and over three 

years the mean optimum densities were 42 and 27 t/ha for 10/12 and 12/14 cm 

grade bulbs, respectively, considerably higher than the planting rates used commercially. 

Figure 4.6 shows the profitability for the one-, two- and three-year 

growing of small offsets and large DN bulbs grown in ridges in south-west 

England at a range of planting densities. Experimental work in Lincolnshire and 

Cornwall indicated that planting rates up to 22.5 t/ha could be profitable where a 

high production of saleable (middle grade) bulbs was required. Higher planting 

densities lead to longer stems and increase crop support on windy sites, so are useful 

where the flower crop is important. For growing narcissus on a one-year-down 

system, Dutch planting rates vary from 14 t/ha for vigorous, small offset planting 

material, to 35 t/ha for large ‘mother bulbs’ where splitting to more saleable grades 

of bulbs is desired. Dwarf narcissus such as Cyclamineus cultivars are usually 

planted at about half the density of standard cultivars.


Figure 4.6 Effect of planting density, bulb grade (offset or double-nosed (DN) bulb) 

and crop duration (1, 2 or 3 years) on profitability. Data for narcissus 

‘Fortune’ grown in ridges, from Rees et al. (1973), with permission from 

The Journal of Horticultural Science and Biotechnology. 

High planting densities may accentuate problems with pests and diseases, create 

handling problems at planting and lifting, and require changes in irrigation or 

fertiliser practices. Planting density has a large effect on the spread of base rot. Linfield 

(1987) placed healthy bulbs 0, 10 or 20 cm laterally from ‘inoculator’ bulbs, 

and after one growing season 60, 27 and 6% of the healthy bulbs were affected, 

respectively. The base rot fungus can infect bulbs 30 cm away (Price, 1975c). 

For all bulbs to be at risk of infection requires 10% affected bulbs at a density of 

10 t/ha, but only 5% at 20 t/ha, on ridges 76 cm apart; lower percentages would be 

required using 90 cm ridges (Tompsett, 1980a). 

Planting depth 

Narcissus bulbs are usually planted about 13 cm deep (from the base of the bulb to 

the top of the ridge). Taking care to adjust machinery to ensure an even planting 

depth, especially on undulating sites, produces a uniform environment and aids 

bulb lifting. Deeper planting may produce better growth, but bulbs are harder to 

lift, while shallower planting may lead to damage from cultivation and herbicides, 

and the bulbs are in warmer soil. Narcissus bulbs have contractile roots, and planting 

bulbs 20 cm deep results in the typical bulb shape, whereas when planted at 5 

or 10 cm deep bulbs become elongated as a result of root action (Tompsett, 1977;

84 G.R. Hanks 

Hanks and Jones, 1986; cf. Allen, 1938). With increasing planting depth (8–23 cm), 

Wallis (1964) reported that emergence and flowering were progressively later, 

while bulb yields were greatest at intermediate depths. Narcissus bulbs can probably 

be planted much deeper than this, although this would be suitable only for 

garden and landscape use, where it would avoid damage from surface cultivation. 

Hagiladi et al. (1992) planted Tazetta bulbs at depths up to 90 cm. Deeper planting 

delayed emergence, and planting deeper than 60 cm resulted in fewer leaves and 

a net loss of bulb yield, although some shoots emerged even from bulbs planted 

90 cm deep. 

Planting arrangement 

When bulbs were planted by hand in beds, they were placed evenly, 1½–2 bulb 

diameters apart and upright in the rows (Rees, 1972). Machine planting tumbles 

bulbs into the planting furrow, so they are more-or-less randomly orientated 

within the ridges, presumably resulting in less uniform growth and shape. The 

effect of bulb orientation was investigated when planting machines were introduced 

(NAAS, 1961; Wallis, 1964). Although random planting produced bulbs 

with bent necks, orientation (vertical, inverted, diagonal or horizontal) did not 

affect bulb yields, although vertical planting gave earlier crops. In the UK, ridges 

are often arranged at distances of 90 cm, centre-to-centre, but 76 cm ridges are 

also used. Planting machinery is often set to plant bulbs in a 20–25 cm wide band 

within the ridge, so the actual planting density within the planted area (the planting 

band) may be 3–4 times the overall (field) area. In trials, the width of the 

planting band (20–35 cm) had little effect on percentage bulb weight increase or 

bulb grade-out with planting densities from 20 to 30 t/ha (Millar, 1978; ADAS, 

1983). In trials with bed-grown narcissus in the Netherlands, changes in planting 

arrangements, from 95 cm planting bands in 140 cm-wide beds, to 105 cm planting 

bands in 150 cm-wide beds, had practical advantages as well as allowing 2% 

more bulbs to be planted in the same area with little impact on labor requirements 

(van Dam and Schaap, 1987). 

Rees et al. (1968) investigated the effects of planting density, rectangularity and 

row orientation in bed-grown narcissus in south-west England. Density did not 

affect anthesis date, but high densities increased stem length by over 20%. Flower 

numbers increased with density, but the number of flowers per bulb fell at the 

highest density. Lifted bulb weight increased with density, but did not peak within 

the range of densities used in this experiment (up to 150 bulbs/m 2 ), although 

yields of larger bulbs reached a maximum above 100 bulbs/m 2 . Bulb yields were 

considered more than adequate, even at the highest planting density. Bulb yield 

declined with increasing rectangularity in east-west rows, but was unaffected by 

rectangularity in north-south rows, probably an effect of wind via shelter or light 

interception. Plants in north-south rows had longer stems. 

In trials comparing ridge- and bed-growing of offsets and double-nosed bulbs at 

54–216 bulbs/m 2 for 1–3 years, also in south-west England, planting at 20–30 t/ha 

gave the highest combined financial returns for bulbs and flowers (Wallis, 1968; Rees, 

1972; Rees et al., 1973). Flower yield in the first year was directly related to density, 

thereafter declining at the higher densities and with larger bulbs. Ridges were 

out-yielded by beds by 26–29%, except at the lowest density where the difference


was 15%. Two- and three-year-down growing was more efficient than one-year 

growing, especially at medium densities. Similar studies in eastern England 

showed that the response to density varied considerably between years, so it was 

difficult to draw general conclusions. 

Pesticide application at planting 

Bulb planting provides an opportunity to apply pesticides directly around the 

bulbs. The main use is for insecticides that prevent the larvae of the large narcissus 

fly entering bulbs, but this method is not currently used because no suitable pesticides 

are available. Until its withdrawal, the persistant insecticide aldrin was used 

successfully in this way in south-west England (Tompsett, 1973). Chastagner 

(1997) reported that most narcissus growers in the Pacific North-West applied a 

nematicide, usually fenamiphos, in-furrow at planting. 

Crop covers and cover crops 

In the UK, covers or cover crops are not used with narcissus, although they have 

been tested on an experimental scale. Despite the possibility of damage by frost, 

sensitive cultivars like ‘Tête-à-Tête’ are now being grown successfully, without covers, 

in England. In the Netherlands, however, it is common to protect narcissus 

and other bulbs by covering the land with straw or reeds, or by sowing a cereal 

crop, after bulb planting. For example, bulbs planted in beds may be covered with 

straw (10 t/ha, or 15–20 t/ha for frost-sensitive cultivars). The straw is removed in 

February, either mechanically or by burning, although lower rates (up to 10 t/ha) 

may be left in place. Alternatively, a lower rate of straw may be used, in combination 

with sowing rye at 250 kg/ha, and the cereal is killed by spraying a contact 

herbicide before the bulb shoots emerge. Where narcissus are grown in ridges, rye 

or barley (250kg/ha) is sown before planting (if sown after the ridges have been 

formed much of the seed falls into the furrows). Rust from plant material used as a 

crop cover may sometimes infect narcissus foliage (Boerema, 1962). 

Trials have been conducted in several countries to evaluate different crop covers 

and cover crops. In the Netherlands, plastic film and a covering of reeds were 

found to be much more effective than straw or rye in insulating bulbs from frost 

penetration (Meijers, 1979). In Danish trials, a number of covering materials was 

shown to increase soil temperatures by 1–2 °C, slightly hastening shoot emergence 

and significantly increasing bulb yields: the materials tested were chopped straw 

(10 t/ha), converted household refuse (100 t/ha), sphagnum peat (130 m 3 /ha), bark 

(70 t/ha) and sawdust (70 t/ha) (Rasmussen, 1976c). In the colder areas of Poland, 

peat, straw, chaff and cow manure have been reported as mulches on bulb crops 

(Dargiewicz, 1971). 

As well as increasing soil temperatures in winter, crop covers reduce soil 

temperatures in summer. Leaving a straw cover in place may reduce base rot by 

decreasing soil temperature in susceptible cultivars like ‘Golden Harvest’ and 

‘Carlton’ (Tompsett, 1986). McClellan (1952) used late-season mulches (straw, foil, 

etc.) in an unsuccessful attempt to reduce soil temperature and losses due to base 

rot, probably because a calculated temperature reduction of 5 °C was needed but 

only a 2 °C drop was attained.

86 G.R. Hanks 

On sloping sites, ground cover crops (cereals, grass or oilseed rape) successfully 

prevented soil erosion if sown immediately after bulb planting and killed before 

crop emergence, provided the growing season was long enough (ADAS, 1987). 

Planting in nets 

The technique of planting bulbs in netting was developed as an aid to bulb recovery 

in heavier soils (Bijl, 1990). In small-scale trials in the UK, the use of netting 

did not reduce bulb yields in ‘Tête-à-Tête’, compared with growing bulbs loose 

(ADAS, 1988b). 

Operations in the field 

This section covers the control of weeds, diseases and pests as well as flower cropping 

or de-heading, irrigation, roguing and inspection, and (for two-year-down 

growing) maintenance of the crop between the two growing seasons. 

Weed control 

Bulb growers generally aim for good weed control in order to prevent competition, 

clean up crops between the two growing seasons, and to assist harvesting 

(weeds can clog lifting machinery). The use of crop covers (such as straw) may be a 

way of reducing reliance on herbicides. 

The effects of weed competition on narcissus yield were examined by Lawson 

(1971, 1976) and Lawson and Wiseman (1972, 1976, 1978). Even when weed 

cover was substantial, weeds had little effect on early spring growth and first-year 

flowering. However, when they resulted in shading during the period of rapid 

bulb growth, leaves and stems grew longer at the expense of bulb yield; shading 

from late-June had no such effect. Narcissus foliage senesced quicker on weedy 

plots, reducing bulb yields, but if weeds were killed late in the season the narcissus 

foliage lodged. Over-wintering weeds that grew up with the narcissus foliage were 

most damaging, producing smaller, less vigorous bulbs. Under weedy conditions 

bulb yield losses approached 20%, or 35% in very dry conditions. Poor weed control 

often relates to failure to control a relatively few resistant species (Wood and 

Howick, 1958; Lawson and Wiseman, 1972). It is possible that over-wintering 

weeds may help the crop by giving winter protection and conserving moisture 

(Lawson, 1971), although this aspect has not been researched. Weeds can also 

lower soil temperature at bulb depth by 4 °C, compared with a weed-free plot, 

which may affect the development of base rot (Tompsett, 1980a). 

Herbicides are used at four stages: (1) contact herbicides are used in autumn/ 

winter before crop emergence; (2) pre-crop-emergence residual herbicides are 

used as late as possible before crop emergence; (3) early-post-emergence residual 

herbicides are used, usually before shoots are about 10 cm tall; (4) a late-season 

herbicide may be used after flowering, although the materials available are 

restricted and application at this stage is difficult because the crop foliage has often 

flopped to shield the soil surface by this time. The post-flowering period is difficult 

for weed control, because the previous herbicide ‘seal’ on the soil surface may be 

broken by the feet of flower pickers, and also because the new flower initials are


being formed at this time and may be sensitive to damage by herbicides; trials have 

taken place on herbicides suitable for application immediately after the flower 

cropping stage, before the crop foliage has spread to cover the furrows completely 

(Briggs and Hanks, 1997). 

Suitable herbicides are given in the standard texts and elsewhere (Mével, 1979; 

ADAS, 1990a), and herbicide trials have been published in several countries (e.g., 

Turquand, 1968; BBLF, 1972; Briggs, 1972a,b; Lawson and Wiseman, 1976; 

Ryan and MacNaeidhe, 1978; Rupasava et al., 1981; Rusalenko et al., 1981; Smith 

and Treaster, 1982, 1984, 1989, 1990; Koster and Kruyer, 1983; Bing, 1985; 

Skroch et al., 1988, 1994; Howard et al., 1990; al Khatib, 1996). Several types of 

herbicide damage can occur, including leaf scorch and chlorosis, abnormalities 

such as distorted flowers, damage to the basal leaf meristem resulting in flaccidity, 

and reduced growth (Ivens, 1966). Cereal seed may be a problem where crops 

have been covered with straw (Koster and de Rooy, 1981; Koster, 1983; Koster 

and van der Meer, 1986). The control of cereal, potato and other ‘volunteers’ 

(plants left from previous crops) in narcissus crops may also prove difficult. 

Fungicide sprays 

It is usual to apply fungicide sprays to narcissus crops to control foliar diseases 

such as smoulder, leaf scorch, fire and white mould, and this is more important 

where bulbs are being grown on a two-year-down basis because of the build-up of 

disease in bulbs, debris or soil. Spray programs are not always successful in controlling 

these diseases (Melville, 1980), although they may help in the control of 

pathogens involved in bulb rots (Davies et al., 1998). A general effect of fungicides, 

especially if a programme of sprays is used, is to delay foliar senescence, probably 

by controlling fungi that degrade the leaf cuticle (Rees, 1972; Jones, 1978). Since 

bulbs are usually lifted before the leaves have died down, this has the disadvantage 

that crops are even greener at lifting. 

There is little specific information on the most effective fungicides or fungicide 

programs for controlling particular diseases, and in the UK it is usual to 

apply several fungicides with different modes of action, which also reduces the 

likelihood of the development of resistance to fungicides. Typical fungicides 

used in the UK and the Netherlands include chlorothalonil, iprodione, vinclozolin, 

mancozeb, zineb/maneb, benomyl, carbendazim, thiophanate-methyl and 

procymidone, sometimes involving tank-mixes. Spraying often begins soon after 

shoot emergence, continuing at 7- to 10-day intervals until flowering, with one 

or two further sprays after flowering to control infections resulting from the 

damage of flower cropping (O’Neill and Mansfield, 1982; O’Neill et al., 1982). In 

practice, it may be difficult to apply fungicides at target dates because of unsuitable 

weather, a particular problem in wet and windy areas where suitable ‘spraying 

days’ may be relatively few. In the year of lifting, the spray programme is 

often curtailed to encourage the foliage to die down. At present, no detailed recommendations 

are available about the critical times for spraying to take place 

(Hanks and Briggs, 1999), but key times are thought to be when frost-damage 

occurs, after flowering or cropping (because of the damage caused by cropping 

or the presence of decaying flowers if not cropped), as well as in periods of damp 

weather.

88 G.R. Hanks 

Insecticide and nematicide applications 

In climates where the large narcissus fly is a problem, it is advisable to apply 

appropriate insecticides if available. Regular sprays against adult flies should be 

applied, preferably making use of pest forecasting models to target applications 

accurately (Finch et al., 1990; Collier and Finch, 1992). Suitable insecticides 

include omethoate (Conijn, 1990; Conijn and Koster, 1990), sprayed just before or 

during the oviposition period. The application of granular and liquid insecticides 

to the ridges during the growing season, to target the newly hatched larvae, has 

also been used. Many insecticides have been tested, but their effects are not always 

consistent each year (Bogatko, 1988; Bogatko and Mynett, 1990; Tones et al., 

1990; Ben-Yarkir et al., 1997). 

Foliar applications of oxamyl were evaluated for the control of stem nematode 

by Westerdahl et al. (1991) as an alternative to HWT with formaldehyde, preplanting 

soil sterilisation with 1,3-dichloropropene, or applying phorate at planting. 

Several rates and timings of oxamyl application reduced nematode levels in 

bulbs and leaves without any phytotoxicity. Earlier, Bergeson (1955) had applied 

three, weekly applications of Systox or other systemic phosphates to narcissus in 

pot trials, and reported that nematode numbers in the leaves and bulb were 

reduced by these treatments, without toxicity at lower rates. 

In the case of high-health status stocks or VT stocks, the regular application of 

aphicides should be considered in warmer weather when populations are high. 

While aphids only rarely colonize narcissus, several common species spread 

viruses during exploratory probings. Frequent applications of anti-feedant insecticides 

(pyrethroids) can be used, although Broadbent et al. (1957) showed that 

systemic insecticides can increase virus spread, possibly by increasing the irritability 

and probing of aphids before death. Alternatively, mineral oil sprays, which 

disrupt normal transmission of virus particles, may be applied, although they are 

not always effective (Mowat et al., 1984). Mineral oil sprays can, however, reduce 

narcissus yield, in one trial by 50% when sprays were applied weekly (ADAS, 

1982c). Some dwarf cultivars, such as ‘Tête-à-Tête’ and ‘Hawera’, appear to be 

more sensitive to mineral oil sprays (Vreeburg and Korsuize, 1987). 

Other chemical treatments 

The application of ammonium nitrate sprays can delay leaf senescence and 

increase bulb yields, perhaps by replacing the failing uptake from senescent roots 

(Rees, 1972). No beneficial effects on bulb yield have been reported from UK trials 

in which a range of plant growth regulators (PGR) were applied in the field (ADAS, 

1984). However, for N. tazetta, El Sallami (1997) reported a range of effects when 

PGR were used as bulb soaks or foliar sprays, including increased bulb production 

with ethephon. 

Flower cropping and de-heading 

Depending on local practices, flowers from narcissus crops are either cropped 

routinely, only when market prices make this worthwhile, or not at all if bulb 

production is paramount. When grown for processing, it is unlikely that flowers


will be cropped because of the physical damage this causes, the likely spread of 

disease, the loss of photosynthetic area and the high labor requirements, and 

perhaps also because of contractual arrangements. Flower cropping may increase 

the incidence of smoulder because of the opportunity for the fungus to invade 

damaged surfaces (Gray and Shiel, 1975, 1987; Dixon, 1985, 1986). 

On the other hand, de-heading crops may increase bulb production by removing 

a sink to nutrients, and by eliminating decaying flowers that may encourage 

fungal diseases. However, the results of trials have been variable. Thus, Wallace 

and Horton (1935) reported instances of 40 and 103% greater yields of bulbs 

when the flowers were not cut, whereas Allen (1938) reported only minor effects 

of flower or flower stem removal on bulb yield. Grainger (1941) found that the 

stem contributed little to bulb growth, and suggested it should be removed. Kalin 

(1954, 1956) reported highest bulb yields after de-heading: yields were reduced by 

3% when flowers were left intact or cropped half-way up the stem, by 5% when the 

flowers were cropped at bud stage, and by 7% when cropped at full bloom. 

Removing the flower bud as soon as the stem had grown enough to allow it gave 

yields similar to those of de-heading. When these practices were repeated annually, 

the effects were cumulative. De Vlugt and Kruijer (1975) confirmed similar 

yield losses as a result of not de-heading (1%) or not picking flowers (5%). The 

differences between different trials may be due to cultural practices, cultivars, location 

or how carefully the cropping and de-heading treatments were carried out, 

although, in the case of de Vlugt and Kruijer’s (1975) study, the results were 

similar whether ‘careful’ or ‘commercial’ standards of removal were used. Some 

cultivars have brittle stems, and large amounts of damage would be expected from 

de-heading (Tompsett, 1976). Removing leaves at cropping, compared with cropping 

flowers alone, further reduced yields by 66 and 36% when only one or two 

leaves were left attached, respectively (Allen, 1938). Overall, de-heading is probably 

not economically worthwhile. The removal of non-cropped flower heads is 

recommended in the Netherlands to control fire, but, although de-heading 

machines have been tested, this is not usually practised (van Aartrijk, 1990). 

Whether flowers are cropped, de-headed or left intact, a fungicide spray programme 

is important for different reasons. 

Irrigation 

Trials have shown that bulb yields are best in soils near field capacity (Strojny, 

1975; Goniewicz et al., 1976). A rise in soil moisture over the range 40–95% of 

available water capacity had no effect on N, P or K levels in the bulbs, but levels of 

P and K in the roots rose with increasing moisture levels (Dabrowska, 1975). 

Water availability also altered root anatomy and stomatal numbers in narcissus 

(Goniewicz et al., 1976). 

In the Netherlands, irrigation of bulb crops in sandy soils is normal, often 

through the control of the water table. It is recommended that narcissus should be 

irrigated when it becomes difficult to squeeze the soil round the roots into a ball, 

and water should be applied in applications of 15–20 mm, as higher applications 

damage soil structure. Narcissus crops are not usually irrigated in the UK, 

although some trials have shown that irrigation increases the yield of larger bulbs, 

especially at higher planting rates (ADAS, 1985d). Moderate irrigation may improve

90 G.R. Hanks 

bulb growth, especially in April-May when rapid growth is taking place, and it also 

improves soil conditions for bulb lifting. Excessive irrigation late in the growing 

season (late-May onwards) may increase bulb weight but may delay the ‘ripening’ 

of the bulb, as well as causing split scales due to uneven growth (ADAS, 1970). 

Benefits of irrigation were demonstrated in trials in New Zealand (McIntosh and 

Allen, 1992). 

Roguing and selection 

When grown as ornamentals it is important to inspect crops and physically remove 

rogue cultivars and other off-types, but whatever narcissus are grown for they 

should be inspected for signs of stem nematode lesions (‘spickels’), disease ‘primaries’ 

(such as smoulder) and virus symptoms. Affected plants should be removed 

and destroyed. Roguing is a skilled and labor-intensive operation: traditionally, 

affected bulbs were dug out with a ‘roguing iron’ inserted into the ridge under the 

bulb. Various methods of roguing using herbicides (e.g., paraquat or glyphosate 

guns, gloves, sticks or aerosol sprays) have been tried (Millar, 1977, 1979; Ryan 

et al., 1979; Bijl, 1981). To reduce the spread of viruses, crops should be inspected 

regularly: although severe infestations may be seen early in the season, some 

symptoms become evident only later. Virus spread is proportional to the number 

of infected plants, so this must be kept at a low level by roguing. In a three-year 

period, 16, 46 and 90% of healthy plants became infected in plots with initial 

infector levels of 10, 20 and 50%, respectively (Haasis, 1939; Broadbent et al., 

1962). Improved stocks (called ‘greenstocks’) can be built-up through vigorous 

roguing and by selecting the largest bulbs (ADAS, 1978) or plants with the desired 

characteristics (Chen et al., 1988). 

Crop inspection 

Where narcissus bulbs are to be sold for growing-on commercially, they may need 

to be inspected and certified by the appropriate plant health authority. In the UK, 

growing season inspections are carried out by PHSI to ensure freedom from stem 

nematode. Checking for freedom from other pests and diseases is the responsibility 

of the grower. 

Operations between growing seasons 

During the summer between the two growing years, crops may be re-ridged to 

maintain good conditions around the bulbs, to remove dried bulb foliage and to 

seal the soil surface. It is important that all crop foliage is removed and the soil 

surface is cultivated to close cracks, so that contact or translocated herbicides 

subsequently applied do not reach and damage the bulbs (de Rooy and Koster, 

1978; ADAS, 1987). It is not clear whether there are implications of re-ridging for 

disease control: Millar (1978, 1979) reported that the effects of re-ridging on neck 

rot and smoulder were variable, and Melville (1980) reported that, although leaf 

debris is a prime source of infection by smoulder, burning debris did not control 

the disease. Waterlogged furrows could assist the spread of stem nematode, which 

can move up to 1 m in a year, so, where the furrows have been, or are liable to


become, waterlogged, it is useful to improve drainage by breaking up the soil with 

a single tine in early-August, when roots will not be damaged. 

Bulb lifting 

Lifting date 

Traditionally, bulb crops were lifted, once, most (95%) of the foliage had died 

down, in July in the UK. This maximizes yield and ensures that bulbs are ‘mature’, 

with well developed outer skins, when lifted. One disadvantage is that bulbs then 

remain in the ground when soil temperatures are increasing, encouraging diseases 

such as base rot: infection occurs late in the growing season, when moribund 

roots are present (Hawker, 1935, 1943). McClellan (1952) reported that infection 

with the base rot pathogen was related to soil temperature, occurring only above 

13 °C and reaching a maximum of 29 °C. Further, when bulbs are lifted at this 

time it may not be possible to dry, clean and grade bulbs and meet sales deadlines 

or produce bulbs for early forcing. Early lifting, overcomes these problems, and 

may be useful in pest avoidance: for example, the larvae of the large narcissus fly 

hatch and invade bulbs late in the growing season (June), while if foliage is 

allowed to senesce naturally, a three-fold increase in virus levels occurs (Mowat, 

1980a). For these reasons, but mainly to have bulbs ready for the export market, it 

is usual in the UK to lift bulbs from early-June onwards, before foliage senescence 

is well advanced, which requires the foliage to be removed prior to lifting. When 

growing bulbs for processing, other considerations may apply, but bulb lifting is 

possible over a window of at least two months. If lifting is delayed when the soil 

is moist, there is a danger of bulbs re-rooting before lifting. Very late bulb lifting 

may also mean that HWT, if necessary, is given well after the ideal date. 

When bulbs are lifted early, yields are reduced because photosynthesis and 

assimilation have been limited, and the choice of lifting date is a balance between 

an acceptable loss of yield and the advantages of early lifting described. After planting, 

total plant dry weight falls until March, and this is followed by a period of 

rapid weight gain. The curve of growth is sigmoidal, with the linear phase extending 

from late-April to early-June (in southern England) (Rees, 1972) (Figure 4.7). 

Yield losses are therefore severe if bulbs are harvested before June, as shown, for 

example, by the data of Allen (1938) and van der Weijden (1987). Very early lifting 

(in May) leads to reduced flower numbers (Allen, 1938; Rees and Hanks, 

1984). Reporting a trial on the date of defoliation carried out with a number of 

cultivars, Kingdom (1981) stated that removing leaves two weeks after flowering 

was very detrimental to yield, after four weeks was adverse but not destructive in 

all cultivars, and after six weeks gave results comparable with those of intact controls. 

In any case, bulbs should be lifted early if they are known to be infested with 

stem nematode, so that they can receive HWT early, and such bulbs should not be 

allowed to dry out before HWT because of the likely formation of nematode 

‘wool’. Early lifting and HWT also improves the control of base rot. 

Where it is desired to maximize yields, and pest and disease considerations are 

not significant, there may be advantages of delaying foliar senescence through the 

continued use of a fungicide sprays programme. The critical factors controlling 

senescence are not known, but it appears to be stimulated by high temperatures, 

even when cooler weather follows (Rees, 1972).

92 G.R. Hanks 

Figure 4.7 Changes in plant dry weight and leaf area index during the growing season 

Data for narcissus ‘Golden Harvest’ from Rees (1972), with permission of 

Academic Press Ltd. 

Foliage removal 

Where bulbs are lifted early, the green foliage is usually removed using acid or 

mechanically. Sulphuric acid (77%) can be applied as a crop spray by contractors, 

working to strict protocols. Crop foliage and weeds are desiccated quickly. There 

are no known harmful effects on subsequent crop performance. Suitable contact 

herbicides for desiccating narcissus, such as dinoseb, are no longer available. 

Mechanical methods of leaf removal (‘flailing’ or ‘top bashing’) remove the foliage 

at a greater or lesser distance above the bulb neck, cutting either above soil level or 

into the soil of the ridge tops (in which case less soil is lifted when harvesting), and 

depositing the excised foliage on the ground. Chain harrows may be used to 

remove foliage from the ridge tops, and this also help break clods before lifting. 

Linfield (1990) surveyed crop husbandry practices in relation to neck rot: the 

main correlation was that neck rot increased when foliage removal involved 

cutting into the ridge, close to the bulb neck, rather than cutting above the ridge, 

and when there was an interval of several days between flailing and lifting. Clearly, 

the presence of damaged shoot tissue contaminated with soil and debris might be 

expected to result in infection of the bulbs. It is common to remove foliage 

immediately in advance of the bulb lifter. 

Bulb lifting 

Bulbs may be harvested in two stages – in which case they are elevated to the surface 

and picked up manually later – or in one stage, using a ‘complete harvester’.


Two-stage lifting requires considerable labor for picking up bulbs, and is used on 

small farms or specialist operations and where it is usual to leave bulbs in rows on 

the soil surface to dry naturally (‘windrowing’), as is the traditional practice in 

south-west England. On small farms bulbs may be collected into trays, net bags, 

or bulk bins. One-stage lifters vary in complexity, from small tractor-mounted 

machines to large lifting machines which may be manned (for picking off clods by 

hand) or un-manned and which deposit bulbs in bulk bins or trailers (Figure 4.8). 

Specialised bulb lifters may be used, or machines designed for lifting potatoes or 

onions can be modified. Effective separation of bulbs and clods is the key element 

in bulb lifters. The machinery should be designed to minimize mechanical damage. 

Specialist machinery is available for lifting bulbs grown in beds on sandy soil. 

Bulb handling for sale or re-planting 

This phase includes any post-lifting fungicide treatment, drying, storage, cleaning, 

grading and inspection, as well as, for re-planting stocks, HWT and associated 

treatment. HWT was described above, under ‘Pre-planting Bulb Treatments’. All 

these operations are key to the control of base rot in susceptible stocks. In trials 

with highly infested stocks, even when no fungicide was applied (at post-lifting or 

in HWT), the introduction of consistent optimum bulb drying and storage 

regimes reduced the level of base rot markedly (Hanks, 1992b, 1996b). 

Ideally, diseased and damaged bulbs should be removed at all stages of bulb 

handling, although this is labor-intensive and impractical in most stages of 

handling in bulk. Diseased bulbs could include those with obvious rots as well 

as dry rotted bulbs (mummified bulbs and ‘puffers’). There are no automated 

methods of detecting and removing diseased bulbs. However, ‘floater-sinker’ 

Figure 4.8 Large unmanned bulb lifter discharging into bulk trailer (Photograph: 

Horticulture Research International).

94 G.R. Hanks 

methods have been tested, in which lighter, infected bulbs generally tend to float, 

albeit with some apparently healthy bulbs (Anon., 1980; Tompsett, 1976); because 

such a procedure could itself spread disease, if used HWT should follow promptly. 

Bulb drying in the field 

Bulbs may be elevated to the surface and left there to dry for several days. In suitable 

situations (in a dry, windy climate) the method is widely used and is successful, 

although there are several disadvantages. Drying is clearly dependent on 

weather, bulbs can be attacked by moulds or can re-root if conditions are damp, 

they are liable to sun scorch, and a reliable supply of labor is needed to pick up 

bulbs. When harvesting bulbs in this way, a post-lifting spray cannot conveniently 

be applied, although some growers are known to spray fungicide over the bulbs as 

they are lifted and deposited on the surface. 

Post-lifting cleaning and fungicide application 

Narcissus bulbs should progress rapidly from the field to drying. When bulbs are 

being handled in bulk containers (½- or 1-tonne bins), rapid transfer to the drying 

area is easy, but it does not allow soil removal, breaking up clumps of bulbs or an 

on-line fungicide application, although bulbs in bulk bins may conveniently 

receive an immediate dip treatment (in formaldehyde) before drying. When bulbs 

are harvested in loose bulk in trailers, they can be unloaded and passed along a 

series of lines, which could involve vibrating riddles and a rotating barrel riddle, 

for separation, soil removal and fungicide spray application en route to the store. 

Mechanised lifting can result in a high clod and stone content, and fluidised bed 

separators may be used to separate bulbs from soil (Zaltzman et al., 1985). Some 

preliminary bulb grading may be needed at this stage in certain operations. 

Narcissus bulbs are not usually washed to remove soil, because of the danger of 

increasing bulb rots, but the method may be used where bulbs are lifted under wet 

conditions from ‘sticky’ soils. 

In the case of cultivars susceptible to base rot, a prompt dip or spray treatment 

is highly beneficial. The benefits of a post-lifting cold dip in formaldehyde have 

been well demonstrated (e.g., Hawker, 1935; ADAS, 1973; Millar, 1978, 1979). A 

fungicide may be added to the formaldehyde in bulb dips. Benzimidazole fungicides 

(thiabendazole, benomyl) were investigated as bulb dips by Gould and Miller 

(1970, 1971a,b), replacing the mercurial fungicide used earlier and which could 

be phytotoxic (Gould et al., 1961; Miller and Gould, 1967). A treatment of 

1000ppm thiabendazole for 30 minutes at 25 °C one day after lifting was found to 

be very effective. Current recommendations in the UK are for a 15 minute dip in 

thiabendazole and formaldehyde at ambient temperatures, within a day of lifting 

bulbs. Dip treatments impose an extra burden on bulb drying and require the 

disposal of spent dips. Spray treatments are more economical, and several fungicides 

are effective used in this way (Hanks, 1994b), but the number of fungicides 

approved for such use is limited; in the UK thiabendazole is used. Thiabendazole 

sprays are usually applied via simple arrangements of conventional spray nozzles 

at a convenient point in the line, but ultrasonic and electrostatic sprayers have 

also been used effectively to give a more even or targeted spray (G.R. Hanks,


Figure 4.9 Letter box drying wall for bulbs in bulk bins. Air is blown through the slots 

in the wall into the pallet bases of the one tonne bins which are stacked 

against them (Photograph: Horticulture Research International). 

unpublished data). While single treatments with thiabendazole can be highly 

effective, it is the cumulative effect of treatment over several years that most effectively 

reduces incidence of the disease, and if base rot is severe fungicide can be 

applied both post-lifting and in HWT (Hanks, 1992b, 1996b). 

Bulb drying 

Rapid and efficient bulb drying is essential for good bulb quality and especially for 

controlling base rot, since the pathogen does not spread effectively under dry conditions 

(Hawker, 1935, 1940). The method of bulb drying depends on the scale of 

the bulb-growing operation and on the temperature regime adopted for drying. 

On small farms, bulbs may be handled in small containers (e.g., bulb trays), in 

which case they may simply be stacked in a well ventilated or open shed, or even 

outdoors, allowing sufficient air spaces between stacks of containers; alternatively, 

to avoid problems due to unsuitable weather and to speed drying, the trays may 

be dried under ceiling fans in a shed or controlled temperature store. Loose bulbs 

can be elevated onto the floor of a bulk store fitted with air ducts for drying as 

used for grain or onions. Bulbs in bulk bins require a special drying facility, a 

‘letter box’ drying wall (Figure 4.9). The bins have solid sides and a slatted pallet 

base, of which the fork-lift slots form the air duct for drying and ventilating bulbs 

via the slatted base of the bin. The bins are placed against slots in the drying wall 

located at appropriate positions to match the pallet bases of the bins, building up a 

line of bins of length and height appropriate to the facility, and closing the pallet 

bases at the outer ends of the rows with a temporary closer, usually a thick piece of 

foam rubber, so that the air flow from the letter boxes is forced upwards through

96 G.R. Hanks 

the boxes, exiting at the top of the stack. Small numbers of bins can be dried using 

portable fans blowing into the pallet base or using ‘box tops’ fitted with fans 

blowing downwards, or by placing bins over flow ducts in a bulk drier, but in 

either case an arrangement of foam rubber closers or polythene film fixed around 

the bins is needed to direct the air flow through the bulbs. 

The forced air used to dry bulbs may be at ambient temperatures or it may be 

heated. A number of recommendations state that a lift of about 3 °C at temperatures 

of about 25 °C should be used (e.g., van Paridon, 1990), contrary to usual 

advice to growers in the UK (e.g., ADAS, 1988a). Price (1975a,b) showed that the 

incidence of rotting bulbs, and the numbers of propagules of the base rot pathogen 

isolated from the base plate of healthy bulbs, increased with increasing storage 

temperatures from 15 to 24 °C, then declining to 30 °C. In early studies of base rot, 

Gregory (1932) and Hawker (1935) showed that bulbs should be stored below 

25 °C, while Xu et al. (1987) reported that the incidence of base rot was greater with 

temperatures >19 °C and Moore et al. (1979) stated that storage at 18 °C is a reasonably 

acceptable and practical recommendation. ‘High temperature drying’ of 

narcissus bulbs at 35 °C has been developed in the UK, and, as well as the convenience 

of rapid surface drying (in two to three days), it produces cleaner bulbs as the 

outer skins and soil contamination are more easily removed, and there is no 

increase in base rot due to the higher temperature (Tompsett, 1977). However, the 

safety of drying narcissus bulbs at 35 °C has been questioned by Linfield (1986b) on 

the basis of culture experiments with the base rot pathogen on solid and liquid 

media. On solid media, the fungus grew rapidly at temperatures of 20 or 25 °C, but 

growth was slower outside this range and had ceased at 40 °C, confirming the findings 

of McClellan (1952) that the optimum temperature for growth was 24 °C and 

that there was little growth at 35 °C. In liquid media, however, Linfield (1986b) 

found that growth of the pathogen was rapid over the range 15–35 °C, and had not 

ceased entirely even at 45 °C, and she argued that in a freshly-lifted bulb, conditions 

would be more like those of liquid culture, so that warm air drying would, 

initially, favour pathogen growth: the rate of moisture removal from the tissues 

would be more important than the drying temperature itself. 

Bulb drying can be divided into first and second stages (Moore, 1980). Where 

high temperature drying at 35 °C is used, this is only for first-stage drying, and 

lower or ambient temperatures are used for second-stage drying. First-stage drying 

consists of the removal of surface water, and is essential for the control of 

surface moulds and other fungi. The rate of loss of surface water depends on the 

rate of air movement and its temperature, and high rates of air movement are 

necessary (425 m 3 /h/t for bulbs in loose bulk, and up to three-times this, for bulbs 

in bulk bins to allow for leakage). With lower air flows, bulbs in the base of the 

stack dry faster than those at the top, subsequently leading to variations in bulb 

performance. Relative humidity should not exceed 75%. Second-stage drying 

extends to the removal of internal water and ensures bulbs are thoroughly dry; 

high rates of air movement are not needed (170 m 3 /h/t for loose bulbs) and the 

humidity can be 80–85%. Higher ventilation and circulation rates are needed 

throughout storage for disease-prone cultivars such as ‘Tête-à-Tête’. Robertson 

et al. (1980) developed a computer simulation of drying times based on air flows, 

temperature and bed depth, which was in reasonable agreement with experience 

in practice. Bulbs may lose 20–25% of their lifted weight during drying, cleaning,


etc., and it is important that drying does not continue to the point of excessive 

weight loss or desiccation. 

Bulb storage 

Once dry, bulbs are often stored at ambient temperatures in sheds, but it is preferable 

to store them at 17–18 °C: lower temperatures can slow development and can 

render the bulbs susceptible to HWT damage, while higher temperatures favour 

the development of base rot. The ambient summer temperature of the region 

needs to be considered in deciding whether controlled temperature storage is 

likely to be cost-effective. Bulb storage is in effect an extension of second-stage 

drying, and free movement of air around the bulbs is essential in controlling 

moulds and re-rooting. When bulbs are in trays or have already been transferred 

to net bags, no forced ventilation may be needed, but where they are held in bulk 

bins, continued use of fans will be needed, if only for part of the day. During this 

phase, bulbs can be extracted for cleaning, grading, etc. 

Little information is available on the harmful effects of ethylene on narcissus 

bulbs. Hitchcock et al. (1932) reported that concentrations as low as 1.5 ppm 

(0.75 ppm for ‘Paper White’) retarded leaf and stem elongation, while 3 ppm or 

more caused a variety of leaf and bud distortions. Hydrogen fluoride can cause 

leaf scorch as an air pollutant (Spierings, 1969). 

If bulbs are kept at high humidities above 30 °C, soft rot due to Rhizopus species 

can reduce narcissus bulbs to a musty mass. This can occur in the transit of bulbs 

or in propagation during the incubation of ‘chips’ (see below). 

Cleaning, inspecting and grading 

Once dry, bulbs are usually passed along a line involving the removal of loose 

skins and soil by vibrating riddles, brushes and dust extractors, bulb splitting and 

removal of damaged and diseased bulbs by hand, grading over a series of riddles, 

and collection and packing of different grades of bulbs. At all stages of bulb handling 

mechanical damage, which could lead to bruising, infection or poor appearance, 

should be minimised by reducing drops or cushioning surfaces. Trials with 

drops of 25 cm have shown that narcissus bulbs are susceptible to damage in the 

first 10 days after lifting and later, in autumn, and damage was increased by subsequently 

storing bulbs at ambient temperatures rather than at 17 °C (Schipper, 

1971). Mechanical damage to the base plate can result in infection by the base rot 

fungus (Gregory, 1932; Hawker, 1935). Some studies, however, failed to show a 

correlation between simulated mechanical damage and the later development of 

bulb rots (Millar, 1978). 

When grading narcissus bulbs, it is usual to bump bulbs along a series of slotted 

riddles of progressively larger sizes. Because narcissus bulbs are often asymmetrical 

or flattened on one side, round riddles (as used with tulip bulbs, for example) 

are considered unsuitable. Although other grades may be used, narcissus bulbs are 

often graded in 2 cm-wide bands, bulbs of 12–14 and 14–16 cm grades being used 

for sales and smaller and larger grades being used as re-planting stock (some 

cultivars have relatively small bulbs and other grades may apply). Machines for 

counting and weighing bulbs are available. Graded narcissus bulbs for sale are

98 G.R. Hanks 

usually collected in 25 kg lots in nylon mesh bags. After grading, bulb storage 

should continue under the conditions as before. It is usually advantageous to 

organize bulb handling operations so that re-planting stocks move quickly to 

HWT and planting, and bulbs for sales are separated and despatched promptly. 

Where bulbs are being sold for growing on, inspection and certification may be 

needed from the appropriate plant health authority. For example, in the UK samples 

of bulbs are subjected to dry bulb inspection by the Plant Health and Seeds 

Inspectorate to ensure freedom from soil. 

In the case of narcissus bulbs being grown for processing, many of these steps – 

which are aimed at producing visually attractive, healthy flowering bulbs for the 

ornamentals trade – may not be applicable. 

Long-term bulb storage 

In commercial floriculture, long-term storage methods have been developed for 

narcissus bulbs to facilitate transport to the southern hemisphere or to produce 

very late flowers. Both warm and cool storage methods have been used (Beijer, 

1957). Where bulbs are being supplied for processing, longer-than-usual storage 

may be necessary to suit production schedules. 

In retarding narcissus bulbs by warm storage, Dutch bulbs were formerly stored 

at 28 °C and 70% relative humidity from lifting (in July) until shipping the following 

year and planting in South Africa in August (Hartsema and Blaauw, 1935). 

Long-term storage at extreme temperatures (–1.5 °C or 34 °C) was detrimental to 

the bulbs, but good results were obtained by storage at 25.5 to 31 °C followed by 

10 weeks at 17 °C (Hartsema and Blaauw, 1935). Beijer (1957) obtained best 

results by storage at 30 °C from lifting to mid-October, followed by –0.5 °C until 

late-December then 25.5 °C until shipping in February-March and planting in 

April. For longer storage, 25.5 °C was used from late-November (Beijer, 1938). 

Warm storage can be prolonged for a year, making all-year-round flowering 

possible. This has been investigated with several standard cultivars (ADAS, 1970, 

1989a; Tompsett, 1988). Bulbs were stored for several months from lifting at 26 °C 

and 70% relative humidity, then at 17 °C for 4 weeks before being planted and 

placed at 9 °C for 6 weeks; after this they were transferred to cool growing conditions. 

The normal cold requirement did not seem to apply, but the 9 °C period 

promoted root growth. The required flowering dates could be attained by ‘holding 

back’ the plants at 2–5 °C as necessary. 

Long-term storage of narcissus bulbs at low temperatures was investigated by 

Griffiths (1936). Bulbs were stored at 1 °C from November for up to 10 months. 

Vegetative growth was satisfactory, although the flower buds had died, indicating 

that prolonged cold storage for non-cropping purposes should be satisfactory. 

Beijer (1957) reported that bulbs could be shipped in September following storage 

at 20 °C, then planted in November at 5 °C for 1 month and then at 0 °C until as 

long as July, flowering satisfactorily. Alternatively, bulbs can be frozen at –1.5 °C 

over winter and spring, planted and then stored at 7 °C for 3 weeks followed by 

9 °C for 2 weeks and then 1 °C as required, flowering thereafter taking place in 

4–6 weeks (ADAS, 1989a). 

Transient exposure to sub-zero temperatures, during growth or after harvest, 

damages many flower-bulbs. Cohen et al. (1997) investigated the hardening of bulbs


of Narcissus tazetta ‘Ziva’ to freezing stress. Acclimatisation of bulbs by hardening at 

2 °C was unsuccessful, but a single soil drench with paclobutrazol or uniconazole 

produced daughter bulbs that were not injured by freezing at –2 °C for 12 hours. 

Transport 

For shipping bulbs, well-ventilated refrigerated containers (reefers) should be 

used, preventing losses due to ‘heating in transit’ observed in earlier years. On 

receipt, bulbs should be stored in well-ventilated, ethylene-free stores at 13–17 °C 

(De Hertogh, 1989). De Hertogh et al. (1978) investigated hypobaric storage as an 

aid to shipping. Bulbs were stored for 2 weeks at 76 mm Hg and 17 °C prior to 

cooling: low pressure storage retarded bud growth, but eventual flowering was not 

affected, compared with control bulbs stored under ambient conditions. 

SPECIALIST TYPES OF NARCISSUS BULB PRODUCTION 

As well as the ‘standard’ narcissus and daffodil cultivars, other types of narcissus 

may be required, needing different growing techniques. These include Tazetta 

narcissus, dwarf and small-bulbed cultivars and Narcissus species. Integrated crop 

management and organic systems of growing are also considered. 

Production of Tazetta Narcissus 

Tazetta narcissus require a frost-free climate for natural-season growing. The 

main producing country for Tazetta bulbs is Israel (Yahel and Sandler, 1986; van 

der Weijden, 1988). Here, bulbs are planted in October, lifted in June and stored 

at ambient temperatures (25–30 °C), which retard the bulbs naturally until temperatures 

fall low enough to allow growth. Floral initiation takes place after lifting, 

in July/August, and anthesis occurs before winter if other conditions are favourable. 

The bulbs can be retarded by storage at 30 °C, producing late flowers in April or 

May (Yahel and Sandler, 1986). In northern Europe, a suitable climate exists in 

the Isles of Scilly, where the Tazetta cultivar ‘Grand Soleil d’Or’ (‘Sols’) is an 

important crop, although bulb production here is secondary to the production of 

the early, fragrant flowers (Veldt, 1988; Schaap, 1989). Cultural methods are 

usually adapted from standard practices for the region (ADAS, 1970), but are 

largely influenced by the local requirement to extend the flower cropping season 

and to maximize flower, rather than bulb, yields. These procedures include leaving 

crops down for several years (Tompsett, 1980b), early lifting and heat treatment 

(Rees and Goodway, 1970), burning-over using a tractor-mounted propane 

burner, and covering the crop with polythene film (Tompsett, 1980b, 1985). 

Similar responses have been reported for other Tazetta cultivars, including 

‘Paperwhites’ (Imanishi, 1983; Tompsett, 1985). Although bulb production from 

Israeli bulbs in the Isles of Scilly is satisfactory in the first year, growth in the 

second year is poor, because higher temperatures are needed (Vreeburg and 

Korsuize, 1989). In unsuitable climates, or to enhance growth, Tazetta narcissus 

can be grown under protection (Kim and Lee, 1982). As well as being unusual in 

not having a cold requirement, narcissus of the Tazetta group characteristically

100 G.R. Hanks 

respond to ethylene or smoke treatments with faster or better flowering (Imanishi, 

1983; Imanishi and Ohbiki, 1986). 

Tazetta bulbs can be converted to summer crops to provide a means of bulb 

production where they are not naturally hardy (Tompsett, 1988; ADAS, 1989a). 

This involves warm-storage techniques for retarding flowering: bulbs were stored 

at 30 °C over winter and then for 4 weeks at 25 °C before planting outdoors in 

March, producing satisfactory yields in south-west England when bulbs were lifted 

in late-October. The technique was also used successfully in eastern England (G.R. 

Hanks, unpublished data). In the Netherlands, bulbs imported from Israel were 

stored at 30 °C from receipt until planting in spring. The best bulb yields were 

obtained when bulbs were planted in April to May and harvested in late-October 

(van der Weijden, 1988; Vreeburg and Korsuize, 1989; Vreeburg and Dop, 1990). 

Bulbs could be stored at 2 °C instead of 30 °C, but prolonged cold storage resulted 

in damage to the leaves. 

While Tazetta narcissus are resistant to base rot, serious losses have been 

reported in Israel due to the nematode Aphelenchoides subtenuis (Mor and Spiegel, 

1993). The nematode infects the roots and secondary infections of fungi such as 

Fusarium cause bulb rotting, giving the syndrome the name ‘basal plate disease’. In 

the Isles of Scilly the nematode Pratylenchus penetrans can attack narcissus bulbs, 

leading to a ‘root rot’ in conjunction with the fungus Nectria radicicola. 

Production of dwarf and small-bulbed cultivars and Narcissus species 

The importance of dwarf and small-bulbed narcissus, such as Cyclamineus, Jonquilla 

and Triandrus types, was referred to in the section on production statistics. Many 

of these types have small bulbs, requiring the use of sandy soils to facilitate bulb 

lifting, and requiring more labor-intensive bulb handling generally. It may be 

appropriate to modify equipment designed for handling small bulbs like freesias 

or onion sets. Many are relatively ‘delicate’ or, like ‘Tête-à-Tête’, are prone to 

diseases such as Penicillium rots, skin diseases or smoulder (van der Weijden, 

1989). The production of these types therefore requires extra care in pesticide use 

and in drying and storing bulbs, and examples have already been cited in the 

section on the production of standard narcissus bulbs. 

There is very little commercial production of Narcissus species and it is limited to 

specialist nurseries, but bulbs of many species have been exported from Mediterranean 

countries, especially Portugal (Oldfield, 1989). Several Narcissus species are 

considered to be under threat as a result of over-collecting or loss of habitats 

(Koopowitz and Kaye, 1990). Commercial bulb companies are now very aware of 

the environmental implications of trading wild-collected bulbs, and, because of 

consumer interest in these attractive species, there would be scope for commercial 

production if appropriate, sustainable farming methods and stocks were available 

(Hanks and Mathew, 1997). 

Integrated crop management and organic production 

At the present time the demand for more ‘environmentally friendly’ or ‘organic’ 

production of ornamental crops is in its infancy, but is probably inevitable that it 

will increase, following trends in food crops, particularly as multiple retailers


promote more ‘sympathetic’ production protocols. This might also apply to plants 

being grown for processing for the production of pharmaceuticals, where it may 

be desirable to exclude pesticides for various reasons. As well as environmental 

reasons for encouraging less reliance on pesticides, there is the practical situation 

that relatively few pesticides are approved for use on horticultural crops: sales of 

pesticides, other than for use on major crops, are unlikely to justify the development 

and registration costs, and the chemical armoury of the grower of horticultural 

crops has decreased in recent years and may decrease further. In the 

Netherlands, the major bulb-growing country, the intensity of horticultural 

production, high use of pesticides and fertilisers to reduce losses due to pests and 

diseases and to increase yields, and the vulnerability of the water table and water 

courses have led to major restrictions on the use of agrochemicals in the bulbs 

industry. Raven and Stokkers (1992) and Stokkers (1992) summarised this situation, 

reporting that 10% of the use of pesticides in the Netherlands was used in 

the production of flower-bulbs (a 12-fold higher input than the average input per 

ha), and listed the objectives of the Dutch ‘Multi-year Crop Protection Plan’ 

(Anon., 1990). There is a need to develop integrated crop management (ICM) for 

narcissus crops. The setting up of experimental farms to test prototype ICM systems 

for bulbs was described by Raven and Stokkers (1992) and De Vroomen and 

Stokkers (1997). De Ruijter and Jansma (1994) described a model which optimizes 

production as regards environmental goals (nitrogen residues and pesticide inputs) 

and financial goals (income) for crops including narcissus, while Rossing et al. 

(1997) explored the options for environmentally friendly flower-bulb production 

systems and the prospects for pest and disease control in bulb crops in a world less 

dependent on agrochemicals was reviewed by Van Aartrijk (1997). A recent review 

of three systems for growing narcissus in the Netherlands – integrated, experimental 

integrated and biological – suggested there were good prospects for the 

integrated system (Wondergem et al., 1999). 

Up to now, little research has been conducted specifically on more environmentally 

friendly narcissus bulb production, although several possibilities are evident 

from previous R&D (Hanks, 1995b). These include physical, cultural and biological 

methods. Thus, in handling narcissus bulbs, emphasis has been laid on rapid 

drying and correct storage to reduce the levels of base rot (Hanks, 1992b, 1996b). 

Cultural methods of base rot control would include early lifting and late planting 

to avoid high summer soil temperatures, and there is also scope for using mulches 

or controlled weed growth to reduce soil temperatures, conserve water and 

reduce the reliance on herbicides. Koster et al. (1997) reported trials on the development 

of low-dose herbicide treatments for bulbs, involving leaving straw 

mulches in place to prevent weed germination, covering the soil with intercrops 

between bulb crops, and optimising the use of mechanical weed control, which 

might involve changes to bulb planting systems. In trials to control the nematode 

Pratylenchus penetrans, flooding was an effective alternative to soil sterilisation (van 

Beers, 1990). On a small scale, solar sterilisation may be useful (Higgins, 1999). 

The biological control of the base rot pathogen by antagonistic fungi has been 

reported by Langerak (1977), Beale and Pitt (1990, 1995), Hiltunen et al. (1995) 

and Hanks and Linfield (1997). Non-pathogenic micro-organisms (Penicillium 

species, Trichoderma species, Minimedusa polyspora and a Streptomyces species) inhibited 

pathogen growth, reduced disease development, or improved the effects of


using thiabendazole alone. Nematode levels in soils can be reduced by growing 

Tagetes and other species. In experiments in soils inoculated with Pratylenchus penetrans 

or Trichodorid nematodes, the population of P. penetrans was reduced by 

planting Tagetes patula, and bulb yield in a subsequent narcissus crop was 

increased (Conijn, 1994). 

BREEDING AND PROPAGATION 

Breeding Narcissus cultivars 

Fernandes (1967) defined two sub-genera within Narcissus, Hermione with a base 

haploid number of chromosomes of 5 (or 10 or 11), and Narcissus with 7 (or 13), 

and crosses between the sub-genera result in a range of chromosome numbers. 

Brandham (1986, 1992; see also Kington, 1998, p.12) and Brandham and Kirton 

(1987) have made extensive studies of the cytogenetics of narcissus: diploid, triploid 

and tetraploid cultivars are common, with high ploidy levels in some species 

(e.g., N. bulbocodium is hexaploid). Brandham (1992) tabulated data for 731 cultivars. 

Most narcissus cultivars, interpreted as the optimum level of horticultural 

fitness, were tetraploids (2n = 28) (Brandham and West, 1993). Other cytogenetic 

studies of narcissus include those of Kalihaloo (1987), Kalihaloo and Koul (1989) 

and Gonzalez-Aguilera et al. (1988). 

The bulk of narcissus breeding has been carried out by enthusiasts with the 

show-bench in mind, for example, in Northern Ireland, the USA, New Zealand 

and Australia. Commercial bulb growers identify new cultivars that may have the 

right characteristics for commercial exploitation, such as high rates of bulb and 

flower production. De Hertogh and Kamp (1986) and De Hertogh (1990) listed 

the desirable characters for commercial cultivars, such as sturdy stems and leaves, 

reliable bud opening, long-lasting flowers, fragrance, tolerance or resistance to 

diseases, and critical weights for floral initiation such that a double-nosed bulb will 

reliably produce two flowers. In some cases, breeding programs may have more 

specific aims. In the UK, there have been breeding programs aimed at producing 

yellow trumpet and large-cup flowers similar to ‘Golden Harvest’ or ‘Carlton’ but 

with relative resistance to base rot derived from ‘St. Keverne’. One programme, 

concentrating on the production of early field-grown flowers, has already resulted 

in several cultivars being commercialised (Pollock, 1989). In another programme 

investigating the genetic basis of resistance to base rot (Bowes, 1992), new cultivars 

are presently under evaluation (Bowes et al., 1996). The latter programme exploits 

the absolute resistance to base rot found in species such as Narcissus jonquilla 

(Linfield, 1986a, 1990, 1992a,b), rather than the relative (or field) resistance of 

commercial cultivars such as ‘St. Keverne’ (Tompsett, 1986). Breeding for resistance 

to base rot is hampered by the difficulties in screening seedling bulbs because of 

the development of adult plant resistance (Linfield and Price, 1986). Breeding for 

resistance to base rot currently utilizes a lengthy screening method that requires 

large numbers of clonal two-year-old bulbs (Bowes et al., 1992). An in vitro assay 

using bulb scales is showing promise as a much faster alternative (J.H. Carder, 

personal communication). Narcissus pollen can be stored long-term in liquid nitrogen 

(Bowes, 1990).


Mutation breeding of narcissus was reported by Misra (1990). Two narcissus 

cultivars flowered early without leaves, after exposure to gamma radiation. Rahi 

et al. (1998) carried out experiments with N. tazetta ‘Paper White’ in which bulbs 

were exposed to gamma radiation and then planted in normal or alkaline soil. 

The performance of irradiated plants in the alkaline soil indicated possibilities for 

selecting salt-resistant strains. 

D.O. Sage (personal communication) is developing a transformation system 

for narcissus as a possible route to cultivar development. Transgenic callus of 

narcissus ‘Golden Harvest’ has been produced, carrying a selectable marker and 

reporter gene, and attempts are being made to regenerate plants from it. Work 

will then concentrate on ‘clean’ transformation technologies for producing transgenic 

narcissus ultimately without selectable marker and reporter genes. The technology 

should then be able to approach pest and disease control by introducing 

resistance genes to otherwise acceptable cultivars. 

Future uses of narcissus plants may require breeding for characteristics such as 

alkaloid or essential oil content, which have not apparently so far been attempted. 

Whatever the goals of narcissus breeding, there is a need to conserve genetic 

material for future use. Because of the huge numbers of commercial cultivars 

there is a danger that historical but valuable parent cultivars may be lost, while the 

loss of wild species (and potentially useful subspecific taxa) through over-collecting 

or habitat destruction has already begun (see Chapter 3, this volume). Historic 

cultivars and wild types need to be conserved. Koopowitz (1986) has discussed 

the wider implications of conserving amaryllids, including the need for a large 

number of each to represent the variation of the gene pool meaningfully, long 

generation times, specialised cultural requirements and the widespread occurrence 

of virus diseases. 

As well as improving narcissus cultivars, narcissus genes may be useful in the 

production of other transgenic plants. Booth (1957, 1963) studied carotenoids in 

narcissus, finding the coronas to be among the richest sources of carotene. Rice 

contains neither β-carotene (provitamin A) nor its precursors, and Burkhardt et al. 

(1997) transformed rice by microprojectile bombardment with a cDNA coding for 

phytoene synthesis from narcissus. 

Narcissus propagation 

A major problem with commercial narcissus breeding is the long time – 15–20 

years – needed to build up adequate stocks beginning with one bulb and using 

natural multiplication, about 1.6-fold per annum, by which it takes about 16 years 

to go from one to 1000 bulbs (Rees, 1969). Micropropagation is almost certain to 

be required, perhaps followed by a low-cost macro-propagation method, such as 

chipping combined with optimised field production, once reasonable numbers of 

bulbs have been produced (Hanks and Rees, 1979). Propagation from seed is also 

considered. 

Micropropagation 

Whereas macropropagation techniques such as chipping are simple and require 

minimal facilities, in many cases they will not produce the multiplication rates


required for the effective bulking of a stock in a reasonable period. Further, 

meristem-tip culture can be used to produce ‘virus-free’ plants (Stone, 1973; 

Phillips, 1990; Sochacki et al., 1997). 

Hussey (1975, 1980, 1982) reported the regeneration of plants using leaf, scale 

and stem explants and from callus derived from the ovary wall. Using scale tissue, 

usually double scale segments (‘mini-chips’), shoots were produced on the abaxial 

surface close to the base plate, and the multicellular origins of such growth suggested 

that the progeny should be genetically uniform. Sub-culturing at 16-week 

intervals, 500–2000 bulblets could be produced from one parent bulb in 18 

months. All shoots eventually became senescent and formed dormant bulblets. 

After a cold treatment of 8–10 weeks at 2–9 °C, bulblets sprouted and could be 

planted out, reaching flowering size in three or four years. Seabrook et al. (1976) 

and Seabrook and Cumming (1982) described shoot, root and callus induction on 

leaf base, stem and ovary segments. The most productive explants were the bases 

of young leaves from cold-treated bulbs, and from two leaf sections, 2620 shoots 

were obtained after five months and four sub-cultures (Seabrook et al., 1976). 

Several cultivars were tested and were found to differ in vigor, but multiplication 

rates of up to 27-fold per annum were achieved. However, the plantlets transferred 

to soil with difficulty. 

Squires and Langton (1990) carried out an evaluation of narcissus micropropagation 

on a commercial scale. Using an adaptation of Hussey’s (1982) methods, 

shoots were obtained from ‘mini-chips’ and sub-cultured before senescence was 

induced by transfer to a hormone-free medium; after a cold treatment, bulblets 

were planted out. There were large differences between cultivars, but, on average, 

7.6 shoots were obtained initially from each bulb, there was a multiplication rate of 

1.8 per transfer, a rate of conversion to bulblets of 1.4, and 60% success in transplanting. 

From one parent bulb, starting in August and using only the inner scales 

(to reduce contamination), using nine transfers and planting out after 18 months 

for three or four years to produce flowering size bulbs, 1200 bulbs were produced, 

with a labor requirement of 2.6 man-minutes per bulb. Although the 

multiplication rate was lower than that of Seabrook et al. (1976), planting out bulblets 

rather than rooted shoots was more convenient, and a high degree of genetic 

stability was expected because the shoots produced had an origin similar to that of 

natural increase. Squires et al. (1991) investigated the causes of poor transplanting, 

and adjustments to the culture method improved the success of transplantation to 

81% in some cultivars. The main constraints to better growth rates were the onset 

of leaf senescence both in culture and in the year after transplanting. Bulblet 

weights of 0.20 g were needed for reliable transplanting. 

The micropropagation of narcissus was also investigated by Selby and co-workers 

(for review, see Harvey and Selby, 1997). In narcissus micropropagation, a large 

amorphous mass of achlorophyllous tissue develops at the base of the shoot-clump 

cultures and new leaves arise from it (Chow, 1990). Chow et al. (1993) and Harvey 

et al. (1994) compared the structure of the basal tissue of the shoot-clump cultures 

with the base plate of bulbs, and showed they were similar. Thus the leaves of 

such cultures do not appear to derive from callus and therefore should be trueto-type. 

Non-dormant bulbils that transferred to soil well could be produced 

from shoot clump cultures by using high sucrose levels in culture (Chow et al., 

1992a). Multiplication rates in culture could be increased by trimming all green


leaf tissue at alternate transfers (Chow et al., 1992b). Staikidou et al. (1994) compared 

bulbil formation by single leaf explants and by shoot clump cultures. Bulbil 

formation on the former was slow and inefficient compared with using shoot 

clump cultures, but was responsive to growth regulators and so might be a useful, 

simple system for investigating the regulation of narcissus bulbil initiation and 

development. 

Other reports on the micropropagation of narcissus include those of Popov and 

Cherkasov (1984), Paek et al. (1987), Kozak (1991), Langens-Gerrits and Nashimoto 

(1997) and Sochacki et al. (1997), who used various explants including those 

consisting of scale bases with attached base plate tissue. Infection of cultures with 

Fusarium is a problem in the tissue culture of narcissus, and Hol and van der Linde 

(1989) demonstrated that infection could be controlled by storing bulbs at 30 °C 

from lifting and giving bulb HWT at 54 °C for 1 hour before setting up cultures. 

Riera et al. (1993) reported the beneficial effect of the polyamine diaminopropane 

on regeneration from twin-scales in culture. Langens-Gerrits and Nashimoto 

(1997) demonstrated that the presence of roots on bulblets was needed to obtain 

good subsequent growth when they were planted out. Joung et al. (1997) reported 

regeneration from different parts of the flower stem. Santos et al. (1998) reported 

tissue culture starting with twin-scale explants of N. bulbocodium, in which 90% of 

the bulbs obtained flowered during the first growing season. There are some 

advantages in using a shaken liquid culture over using solid agar media. Bergoñón 

et al. (1992) described shake cultures of N. papyraceus (see also Chapter 7, this 

volume). In other tissue culture investigations with narcissus, ovule culture produced 

viable seeds (Lu et al., 1988), as did excised placentae pollinated in vitro 

(Balatková et al., 1977). 

For Tazetta cultivars, successful induction of plantlets has also been reported 

using young stem explants (Hosoki and Asahira, 1980), stem and scale explants 

(Nagai, 1999a,b) and cultured twin-scale bases (Dachs et al., 1979; Steinitz and 

Yahel, 1982; Gu and Gao, 1987), and Li and Tang (1982) reported callus induction 

on twin-scale segments. In the case of cultivar ‘Grand Soleil d’Or’, under the 

most appropriate conditions 80–100 explants were obtained from each parent 

bulb, giving 200–300 bulblets in 6 months, and 80–90% of bulblets weighing over 

0.25 g with two leaves and abundant roots were transplanted successfully (Dachs 

et al., 1979; Steinitz and Yahel, 1982). Gu and Zhang (1991) compared production 

from micro-propagated Tazetta narcissus derived from callus cultures with naturally 

produced bulblets. Micropropagated plants flowered after 4 to 5 years, and 

there were no differences between them or naturally derived plants in the number 

of flowers or in flower morphology. 

Sage et al. (2000) produced somatic embryos from narcissus cultivars. Somatic 

embryos were obtained from leaf lamina, leaf base, bulb scale and, especially, stem 

explants. Leaf explants from shoot cultures produced somatic embryos and 

converted to plantlets efficiently on an appropriate medium following a cold treatment, 

the plantlets subsequently transferring to ex vitro conditions readily. 

Chipping, twin-scaling and related propagation techniques 

Several simple propagation methods suitable for on-farm use have been investigated 

for the multiplication of choice cultivars and VT stocks of narcissus. Scooping


and cross-cutting, standard techniques used with hyacinth bulbs, gave poor results 

with narcissus (Stone, 1973; Stone et al., 1975), and leaf cuttings were unsuccessful 

(Alkema, 1971b). Twin-scaling and chipping were, however, successfully adapted 

to narcissus from methods used with other Amaryllidaceae in which the bulb is cut 

into a number of segments and the segments divided into single or paired scale 

pieces each with a piece of the base plate attached (Luyten, 1935; Traub, 1935; 

Everett, 1954). Only a small percentage of single scale pieces, but over 80% of 

paired scales, produced bulblets (Alkema, 1970, 1971a, 1975; Broertjes and 

Alkema, 1971). These bulb division methods rely on the destruction of the apical 

dominance of the existing buds, resulting in adventitious bud formation on the 

proximal part of the bulb scale adjacent to the base plate (Hussey, 1975; Grootaarts 

et al., 1981). 

In ‘twin-scaling’, flowering-size bulbs are cut into a number of longitudinal 

segments (often 8 or 16), each with a wedge-shaped piece of base plate; these 

segments are further divided by cutting off the scales in pairs, each with a conjoining 

piece of base plate, and 60 to 100 twin-scales can be cut from one large bulb. 

These pieces are incubated in a moist medium (usually damp vermiculite) for 3 

months at 20 °C, when bulblets develop, usually one per ‘twin-scale’ (Alkema, 

1970, 1975; Alkema and van Leeuwen, 1977). Typically, 80–90% of twin-scales 

from bulblets, and a small percentage rots. When grown-on, the bulblets flower in 

their third or fourth year (Mowat, 1980b). This method was used to multiply VT 

narcissus stocks in Scotland (Sutton and Wilson, 1987). Testing a wide range of 

cultivars, Fry (1978) found that one bulb could be multiplied to between seven and 

41 flowering-size bulbs in 4 years, compared with six by natural increase. Practical 

accounts of twin-scaling include those of Flint and Hanks (1982) and Hanks and 

Phillips (1982). 

While twin-scaling is a simple technique, it is time-consuming and the 

propagules are small and delicate. More robust propagules can be obtained by 

only partly dividing the original bulb segments, giving pieces with a few scales 

each (Everett, 1954; Stone, 1973), or by leaving the original 8–16 segments intact, 

the method known as ‘chipping’ (Flint, 1982) (Figure 4.10). Several practical 

accounts of chipping are available (e.g., Flint, 1984; Vreeburg, 1984a, 1986; 

ADAS, 1985c; Hanks, 1989). As well as being used for on-farm multiplication of 

select stocks and cultivars, chipping produces attractive round bulbs ideal for sale 

in ‘pre-packs’ and with potential for improving predictability, uniformity and 

mechanised handling (Vreeburg and van der Weijden, 1987a; Hanks, 1989). The 

method has been used extensively with the popular dwarf cultivar ‘Tête-à-Tête’ 

(Vreeburg and van der Weijden, 1987a), where 14 to 71% of bulblets flowered in 

their second year, and it is also successful with several Narcissus species (Hanks, 

1987). Although earlier projections of multiplication rates now appear unduly 

optimistic, multiplication rates of 3- or 4-fold per annum (6.5 or 5 years from one 

to 1000 bulbs) seem realistic (Hanks and Rees, 1979; Hanks, 1993). 

The factors affecting the productivity of twin-scaling and chipping have been 

reviewed by Hanks and Rees (1979) and Hanks (1993). In the following account, 

only the more important effects will be mentioned. It is generally recommended 

that bulbs for chipping should receive HWT about a week before cutting, which 

improves results and controls bulb scale mites (Vreeburg and van der Weijden, 

1987a), but HWT immediately before chipping is harmful. Although twin-scales


Figure 4.10 Narcissus ‘chips’ with bulblets at the end of incubation. (Photograph: 

Horticulture Research International). 

may form bulbils when cut at any time of the year, subsequent growth is best when 

propagation takes place using ‘dormant’ bulbs in July or August (Alkema and van 

Leeuwen, 1977; Hanks and Rees, 1978; Vreeburg and van der Weijden, 1987a), 

and this also gives propagules that are ready for planting-out at a reasonably convenient 

time of year. In earlier work, emphasis was on cutting many small twinscales 

or chips per bulb in order to maximize multiplication, for example aiming 

for twin-scales weighing 0.5–0.8 g (Mowat and Chambers, 1975) or cutting 16 

chips per bulb (ADAS, 1985c). However, small propagules take four or more years 

to produce bulbs of flowering size, and recent recommendations have been more 

pragmatic, sacrificing numbers for bulbil size and a quicker production of flowering-size 

bulbs: bulbs of 10–12 cm circumference (weighing about 40 g) are cut into 

eight segments only (Vreeburg and van der Weijden, 1987a; Hanks, 1989). 

Cutting rates can be adjusted to initial bulb size to achieve target bulb weights after 

a year, after which bulb rate increase is independent of cutting method (Fenlon 

et al., 1990). While cutting bulbs by hand is the only possibility in twin-scaling, 

chipping machines are available which can increase throughput to about 0.5 t/day 

(Flint et al., 1984; Zandbergen, 1984). Chipping machines are based on either a 

star-shaped blade operated by a pneumatic plunger or on arrangements of circular 

saw blades fed with bulbs or bulb halves on a conveyor. To avoid spreading 

pests and disease between bulbs, and to minimize other contamination, sensible 

hygiene should be observed when twin-scaling or chipping bulbs, including disinfecting 

blades (Vreeburg and van der Weijden, 1987a). Immediately following 

cutting, twin-scales or chips are treated by soaking in a fungicide, effective materials 

including products based on captan and benomyl, often used in combination 

(Vreeburg and van der Weijden, 1987a; Hanks, 1989; Linfield and Price, 1990).


In practice, moulds associated with bulbs may be difficult to control during 

incubation of the propagules, perhaps because of the presence of fungicide-resistant 

strains (Lyon, 1978) or due to using grossly infected bulb stocks (Hanks, 1989). 

Bulbil production on twin-scales is inherently variable because of the different 

properties of bulb scales from different parts of the bulb (Hanks, 1985). 

Chips or twin-scales are usually incubated after cutting, usually by mixing with 

damp vermiculite and holding in trays at 20 °C for 12 weeks (Vreeburg and van 

der Weijden, 1987b; Hanks, 1989). These conditions are ideal for fungal growth 

as well as bulbil production. Attempts to scale up the treatment of machine-cut 

chips by using deep crates, or by omitting the medium and controlling humidity, 

gave promising results (G.R. Hanks, unpublished data). In the period immediately 

after cutting, it is important to control chip temperatures which increase as a 

result of wound respiration and the production of heat by the damped vermiculite 

(Hanks, 1989), although the exact temperature and duration of incubation is not 

critical (Hanks, 1986). By the end of incubation, the bulb scales should be largely 

depleted of reserves: bulblets over 10 mm in length grow well, but smaller ones 

often remain dormant (Hanks and Rees, 1979). The propagules are planted in the 

field following the gentle removal of the vermiculite, and in some cases after a preplanting 

fungicide dip (Vreeburg and van der Weijden, 1987c). In the case of 

twin-scales, better growth has been reported following planting in a frost-free 

glasshouse than in an unheated gauze house (Mowat and Chambers, 1977). As an 

alternative to incubating chips, they may be planted in the field directly after cutting 

and fungicide treatment, but chipping should take place in July so that bulbils 

are produced before soil temperatures become sub-optimal, or a polythene mulch 

may be used to raise soil temperatures from planting to emergence (ADAS, 1987; 

Vreeburg and van der Weijden, 1987b). 

In the field, plant growth should be maximised through careful husbandry, for 

example controlling weeds by using non-damaging herbicides or a straw mulch, 

and having a prolonged fungicide spray programme, and by using a low planting 

density (Vreeburg and van der Weijden, 1987c; Hanks, 1989). Planting densities 

quoted include 0.5 million chips per ha or 1–5 t original bulb weight/ha. 

These methods would be useful in bulking supplies of bulbs for extraction, 

perhaps following an initial phase of micropropagation. Chipping is likely to be 

more practical than twin-scaling, as chips are robust, may be cut by machine, and 

are more uniform. 

Optimised production in the field 

Standard bulb production methods are designed to give the most cost-effective 

production of bulbs and (or) flowers, but in the case of valuable bulb stocks it may 

be more important to maximize bulb yields. This can be achieved by greatly reducing 

planting density and by adopting one-year-down growing. Planting density 

should be reduced so that inter-plant competition is minimised, provided there is 

no wind damage. In a trial of optimised bulb production, low planting densities 

and annual planting and lifting were effective, but there was little additional benefit 

of using combinations of foliar feeding, top-dressing, de-heading, irrigation or 

fungicidal sprays (ADAS, 1982c). However, all these methods might be considered 

beneficial in specific circumstances where it is desired to maximize bulb yields.


Production from seed 

Production from seed is, of course, the method used by breeders, and is also 

important for species that form few offsets. Narcissus breeders may sow seed soon 

after collection, either outdoors or in a cold-frame, and germination occurs 

somewhat unevenly in spring. Caldwell and Wallace (1955) reported that, under 

natural conditions, seed of Narcissus pseudonarcissus germinated naturally in 

November or December, while Wells (1989) stated that, when seed is sown outdoors 

in early May soon after collection, N. bulbocodium types start to germinate in 

late-August and germination continues through the winter, with N. pseudonarcissus 

offspring perhaps not germinating until early spring. Under natural conditions, 

summer drought may induce dormancy in N. pseudonarcissus (Barkham, 1980). 

Rees (1972) stated that narcissus seeds have a cold requirement of many weeks, 

and Linfield and Price (1986) germinated seed of commercial cultivars using a 

cold treatment of 12 weeks at 12 °C. Thompson (1977) reported that, in N. bulbocodium, 

conditioning imbibed seed for 7 weeks at 26 °C (but not at 6 or 16 °C) gave 

rapid germination when subsequently moved to 5–16 °C. This suggested that seed 

should be sown in a warm glasshouse and moved to cooler conditions after 2 

months. Hanks and Mathew (1997) reported that synchronous germination could 

be obtained in N. cyclamineus, N. bulbocodium var. citrinus and N. pseudonarcissus by 

keeping imbibed seed at 25–30 °C for 8–12 weeks then transferring to 15 °C. 

There is little information on other aspects of the seed physiology of narcissus, 

although Caldwell and Wallace (1955) reported that seeds of N. pseudonarcissus 

were not light sensitive. Hanks and Mathew (1997) reported that controlling seedborne 

fungi on N. cyclamineus by disinfection, HWT, etc., was difficult. Under 

optimum conditions, it takes 3 years for seed-raised plants of species such as 

N. bulbocodium and N. triandrus to reach flowering size, 4–5 years in other species 

and 7–8 years in some cultivars (Koopowitz, 1986; Oldfield, 1989). This long 

period needed to produce saleable bulbs is a strong disincentive to commercial 

production. Germination and growing systems for the commercial production 

of Narcissus species were investigated by Hanks and Mathew (1997), who showed 

that seedlings could be successful raised in cellular trays for sale as ‘plug plants’ 

or for growing on, a method useful for species like N. bulbocodium that reach 

flowering size quickly. 

CONCLUSIONS 

This chapter demonstrates that the methods for growing and handling narcissus 

bulbs are well documented, but that there remains considerable scope for changes 

in husbandry and other practices that might make the crop more cost-effectively 

produced when growing for processing, or which would enable it to be grown in 

an more environmentally friendly way. Some major research needs are evident, 

such as the need for cultivars resistant to base rot and virus diseases (or their 

vectors), for safer and (or) more effective chemicals (e.g., for bulb disinfection and 

the control of large narcissus fly), and for the development of mechanical or 

robotic bulb handling and sorting to reduce labour inputs. The transformation of 

narcissus bulb production seen in the UK in recent decades has demonstrated


that, with enterprise, much can be achieved, even in the face of sometimes adverse 

economic circumstances. 

The future needs for growing narcissus bulbs for pharmaceutical and other 

processing purposes are uncertain. In 1995, predictions of a vast expansion of the 

UK narcissus area in order to produce bulbs for galanthamine extraction caused a 

stir in the industry (Anon., 1995; Long, 1996). At present there may be options 

both for the extraction of galanthamine from bulbs, and for utilising new developments 

in the synthesis of the compound. But the array of alkaloids present in 

narcissus, and the wide range of potential uses which they have, suggests there will 

be a need for growing narcissus bulbs as an industrial crop for many years to come. 


I would like to thank the many colleagues, cited as ‘personal communications’, 

who helpfully provided information on particular aspects of narcissus production. 

I am grateful to Jim Briggs for his constructive comments on the text. Much of the 

UK research cited was funded by the Ministry of Agriculture, Fisheries and Food. 

I also thank the Horticultural Development Council for permission to quote from 

a number of their projects. 

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Wood, J. and Howick, S.J. (1958) Experiments with herbicides on beds of narcissus and 

tulip 1955–1958. In Proceedings 4th British Weed Control Conference, pp. 112–115. 

Woodville, H.C. and Morgan, H.G. (1961) Lethal times and temperatures for bulb eelworm 

(Ditylenchus dipsaci). Experimental Horticulture, 5, 19–23. 

Xu, T., Cao, R.B., He, G. and Hu, Y.M. (1987) Study on basal rot of narcissus in Zhoushan 

Islands, Zhejiang Province. Acta Phytophylactica Sinica, 14, 93–98 (in Chinese). 

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Yasuda, I. and Fuji, H. (1963) Re-rooting after cutting out new roots on some bulbs. 2. 

In the cases of iris and daffodil. Scientific Reports of the Faculty of Agriculture, Okayama, 21, 

41–47 (in Japanese). 

Zaltzman, A., Schmilovitch, Z. and Mizrach, A. (1985) Separating flower bulbs from clods 

and stones in a fluidized bed. Canadian Agricultural Engineering, 27, 63–67. 

Zandbergen, J. (1984) Vraag naar ronde bollen leidt tot constructie van machine voor 

parteren van narcissen. Bloembollencultuur, 95 (4), 90. 

Zell, R. (1984) Campaign against formaldehyde grows. New Scientist, 103 (1420), 6.

5 Economics of Narcissus bulb production 

James B. Briggs 

INTRODUCTION 

A variety of influences determines the crops produced on any farm, and physical 

factors such as poor soil or difficult drainage exclude growing Narcissus on many 

farms. However, where soils and rotational requirements are favourable, the potential 

financial returns from narcissus bulb production can be evaluated to see 

whether it could be a feasible and economic enterprise. It is necessary to examine 

the profitability of bulb growing relative to other typical enterprises on similar 

land types. A farmer is only likely to enter into a contract for growing a particular 

crop where there is a financial incentive, or if there are perceived to be other 

advantages such as benefit to the rotation or spread of workload. 

The expression of crop profitability in terms of ‘gross margin per hectare’ is 

widespread, and is effectively an indication of the output less the cost of variable 

inputs. As a rough guide, fixed costs on a typical UK arable farm split to one-third 

labour, one-third machinery and one-third other costs. The allocation of fixed costs 

to specific enterprises is more difficult, and is not widely adopted. If done at all, it 

will normally cover the two major elements of labour and machinery costs. Where 

specialist equipment is required, as for bulbs, it is not likely that this crop will be 

included in the rotation, as it will increase overhead costs significantly. If the equipment 

can be utilised for other crops, or if contractors can be used to carry out the 

work, or if equipment can be leased, then the bulb crop may be a realistic option. 

TYPICAL GROSS MARGINS FOR NARCISSUS BULB PRODUCTION 

Gross margins for narcissus growing are shown in Table 5.1, based on 1996 and 

1999 costings for a typical ‘two-year-down’ narcissus enterprise in England (for 

background information see, for example, ADAS (1985a)). These examples refer 

to growing bulbs for the ornamentals industry and, in growing bulbs for processing, 

some circumstances may be different. This evaluation gives gross margins of 

£1922 and £1554 per hectare per annum for 1996 and 1999, respectively (in this 

narrative, the years quoted refer to the years crops were lifted). The following 

assumptions were made in these calculations: 

1 15 t planting stock (bulbs of grade 16 cm) per hectare at £400/ 

tonne. Output = 13.5 t/ha saleable bulb stock at £450/tonne. Initial stock

132 J.B. Briggs 

Table 5.1 Typical gross margin for growing Narcissus bulbs ‘two-year-down’ in 

the UK 

1996 

(£/ha) 

1999 

(£/ha) 

Output (year 2 only) 

Saleable and planting stock 

Variable costs 

Year 1 

12 750 12 075 

Bulbs 6000 6000 

Hot-water treatment 1088 1200 

Fertilisers 116 116 

Sprays (field) 380 374 

Year 2 

7584 7690 


Sprays (field) 401 357 

Post-lifting spray 600 600 

Drying 300 300 

1321 8905 1277 8967 

3845 3108 

Gross margin per hectare per annum 1922 1554 

prices would need to be used in the gross margin for the first cycle. Bulb prices 

rose sharply in the mid-1990s and saleable stock was realising £525–£550/ 

tonne. There has, however, been a levelling out of bulb prices in the mid-1990s 

at £400–£450/tonne. 

2 For the purposes of this exercise, it has been assumed that there were no flower 

sales. It is estimated that, in seven years out of ten, flower sales in the second 

year of 30 000 bunches/ha (ten stems per bunch) at 10 p/bunch (net of labour 

and marketing costs) could be achieved. On an average basis, this would add an 

additional £2100 to the two-year gross margin. This must be weighed against 

the potential loss of yield of saleable bulbs, which may be as much as 10–20%. 

3 In the first year of planting, growers would probably have to buy planting 

stock at open market prices of £500/tonne, which would reduce the gross 

margin on the first crop by £1500. Once the system is established and planting 

stock is available from the harvested yield, the value of the stock is not important, 

as it is comprised of smaller bulbs that would not normally be marketed. 

4 It has been assumed that bulbs will be harvested in July after natural senescence, 

so that chemical desiccation (using sulphuric acid) is not required. 

Further details of the variable costs used in this exercise are given in Table 5.2. 

Bulb grading, if required, would add approximately £1000/ha to these costs. 

Marketing or packing costs, if applicable, would add £40 per tonne sold or £600/ha. 

Most narcissus crops are grown on a ‘two-year-down’ basis in the UK (see also 

Chapter 4, this volume). The results of trials in Lincolnshire and Cornwall in the 

1980s, comparing one- and two-year-down systems, showed that in the UK 

one-year-down growing was not economic. The introduction of a one-year-down

Economics of bulb production 133 

Table 5.2 Variable costs in growing Narcissus (see Table 5.1). All calculations are based 

on 1 ha of crop at 1999 values 

1 Bulbs planted at 15 t/ha @ £400/tonne 

2 Hot-water treatment for 15 t bulbs @ £30.00/tonne + £50.00/tonne 

thiabendazole fungicide for control of basal rot 

3 Fertilisers: year 1 

100 kg nitrogen @ £0.40/kg 40.00 

100 kg phosphate @ £0.43/kg 32.00 

200 kg potash @ £0.22/kg 44.00 

116.00 

Fertilisers: year 2 

50 kg nitrogen @ £0.40/kg 20.00 

4 Sprays: year 1 

Iprodione fungicide (two sprays) 6 litres @ £16.40/litre 98.40 

Mancozeb fungicide 6 kg @ £3.30/kg 19.80 

Chlorothalonil fungicide 3 litres @ £5.50/litre 16.50 

Paraquat + diquat herbicide 8.5 litres @ £5.60/litre 47.60 

Chlorpropham + linuron herbicide 11.2 litres @ £8.80/litre 98.56 

Glyphosate herbicide 5 litres @ £3.52/litre 17.60 

Cyanazine herbicide 5.2 litres @ £14.60/litre 75.92 

374.38 

Sprays: year 2 

Paraquat + diquat herbicide 8.5 litres @ £5.60/litre 47.60 

Chlorpropham + linuron herbicide 11.2 litres @ £8.80/litre 98.56 

Cyanazine herbicide 5.2 litres @ £14.60/litre 75.92 

Iprodione fungicide (two sprays) 6 litres @ £16.40/litre 98.40 

Chlorothalonil fungicide 3 litres @ £5.50/litre 16.50 

Mancozeb fungicide 6 kg @ £3.30/kg 19.80 

356.78 

5 Post-lifting spray of thiabendazole fungicide, 

30 t bulbs @ £21/tonne, for control of basal rot 

6 Bulb drying in store will be required at a 

fuel cost of approximately £10/tonne 

system should be considered only if rigorous disease control is necessary. To achieve 

gross margins from two, one-year-down crops in excess of the gross margin for a 

two-year-down system (with a 150% bulb weight increase), annual bulb weight 

increases of between 70 and 100% have to be obtained from a bulb planting rate of 

25 t/ha (ADAS, 1984, 1985b, 1987a,b). This can be achieved commercially, but, 

given the need for consistently high yields, it would be difficult to do so. Further, 

in the one-year-down system all bulbs are lifted and re-planted each year, and the 

volume of bulbs handled each year is nearly doubled, creating logistical problems 

for most businesses. The use of the two-year-down growing system, therefore, 

became firmly established in the UK. 

ACTUAL GROSS MARGINS 

The figures given above represent ‘typical’ gross margins. Examples of actual gross 

margins recorded by two bulb growers in Lincolnshire, UK, are given in Table 5.3.


Table 5.3 Actual gross margins from two Narcissus growers 

Grower 1 

(£/ha) 

Notes 

a Grower 1, no figure given for hot-water treatment, so a typical figure for 

treatment without thiabendazole has been used for comparability. 

b Grower 2, no figure given for bulb drying, so a typical figure has been used. 

For Grower 1, the figures used are based on 1993, and, for Grower 2, 1995. The 

bulb yields for Grower 2 are particularly low, but 1995 was a dry year when bulb 

yield was severely reduced where irrigation was not available. It is obvious that both 

of these growers are operating under a lower input of pesticides than the one previously 

described, particularly in terms of using fungicide in hot-water treatment. 

However, more intensive systems to guarantee crop quality, particularly for control 

of basal rot, are becoming necessary, and the typical gross margin figures given 

above are based on such a system. The actual gross margins achieved in these 

examples were £2422 and £2195 per ha per annum, respectively. 

THE RELATIVE PROFITABILITY OF NARCISSUS GROWING 

Arable farming, either of combinable crops or mixed cropping, is currently 

carried out in most of the geographic areas suited to narcissus growing. The 1999 

gross margins of the most common crops are given below, expressed as £/ha: 

Winter wheat (feed)* 530 ± 100 

Winter wheat (milling)* 600 ± 100 

Winter oilseed rape* 450 ± 100 

Winter barley (feed)* 480 ± 100 

Spring barley (malting)* 460 ± 100 

Winter beans* 450 ± 100 

Linseed* 400 ± 100 

Maincrop potatoes 2000 ± 1000 

Sugar beet 900 ± 500 

Bulb onions (spring-sown sets) 2090 ± 1500 

Summer/autumn cauliflower 1800 ± 1300 

Grower 2 

(£/ha) 

Output 

Bulbs sold 5910 3770 

Bulbs re-planted 3087 4848 

8997 8618 

Bulbs 3087 3158 

Hot-water treatment 371 a 

440 


Crop sprays 408 361 

Drying 173 190 b 

4153 4228 

Gross margin (£/ha) 4844 4390 

Gross margin per year (£/ha) 2422 2195


These figures give some idea of the high variability in returns for some crops, particularly 

field vegetables and roots. Climatic factors, particularly rainfall, have a 

significant effect on market prices, notably for potatoes. 

Crops marked with an asterisk in the above list are within the Integrated 

Administration and Control System (IACS) of agricultural support. IACS is the 

system for administering farm support payments within the EU. Included in this 

regime is the requirement for an area of arable land to be set-aside from food 

production, in order that support payments can be claimed on both that land and 

on land cropped with combinable crops such as wheat and oilseed rape. These 

support payments are known as Arable Area Payments, or simply area payments, 

and are one of the main pillars of support under the Common Agricultural Policy 

(CAP). The current level of gross margin is heavily reliant on these subsidies which, 

for the 1999 harvest (including an element of agromonetary compensation), were 

as follows (£/ha): 

Cereals 235 

Proteins 340 

Oilseeds 254 

Linseed 456 

Obviously, changes in the level of support will alter the relative profitability of 

such crops. In order to be eligible for these payments, one of the requirements of 

the scheme is that, an area of land must be ‘set-aside’ and not used for food 

production, in return for which a subsidy of around £298/ha is payable. 

FIXED COSTS 

As mentioned previously, the allocation of fixed costs is not often carried out, yet 

different farming systems carry with them different fixed cost structures, particularly 

in the area of labour and machinery costings. Within each type of enterprise 

there will still be considerable variation due to differences in business circumstances 

and efficiency. The ‘top 25% figures’ are frequently presented to give an 

indication of the performance levels of the most efficient businesses. Table 5.4 

(University of Nottingham Rural Business Research Unit, 1998) shows the actual 

results for 1996–1997 and 1997–1998 for costed farms, grouped by cropping type 

and farm size. 

As a long-term trend, root and vegetable farms generate higher gross margins 

associated with high-value crops such as potatoes and sugar beet. However, exceptional 

grain prices, at the same time as a high area payment, gave high levels of 

profitability in 1996. In contrast, by the 1998 harvest, the gross margin for a cereal 

farm was nearer £200/ha, with many farms generating a negative net farm income. 

This trend has continued in 1999. 

Machinery costs are usually higher on mixed cropping farms, with depreciation 

reflecting a higher value of capital investment in specialist equipment. The use of 

agricultural contractors used to be more common on mixed cropping farms, but is 

now an increasing trend on all types of farm, as machinery becomes more expensive. 

The point at which it is cheaper to use contractors differs with cropping type


Table 5.4 Actual costs of ‘medium’ and ‘small’ cereal a and root/vegetable b farms c 

Farm type 1996–1997 (£/ha) 1997–1998 (£/ha) 

Medium Small Medium Small 

cereals root/ 

vegetable 

cereals root/ 

vegetable 

Notes 

a Cereal farms: >50% area in arable crops and 50% in arable crops and >5% in root/vegetable crops. 

c Source: University of Nottingham Rural Business Research Unit (1998). 

d Labour figures exclude farmer and spouse labour. 

and farm size. Rents are usually higher on land capable of growing higher value 

crops. Labour characteristics are not very clear from the examples given, but total 

labour costs are usually higher for mixed cropping. 

COSTS OF BULB HUSBANDRY AND FIELD OPERATIONS 

The field operations required during the growing of narcissus bulbs are similar to 

those for onions or potatoes. A typical sequence for a two-year crop, with standard 

farmers’ costings (£/ha), would be: 

Ploughing 37 

Power harrowing 25 

Fertiliser application (3 × 9) 27 

Planting 100 

Spraying (12 × 9) 108 

Topping (defoliating) 22 

Harvesting 215 

Transporting to store 84 

618 

cereals root/ 

vegetable 

cereals root/ 

vegetable 

Average size (ha) 311 300 69 65 392 369 69 64 

Output 1022 1121 1050 1162 881 987 929 939 

Gross margin 771 804 738 895 613 680 600 673 

Contract 23 25 28 32 20 23 31 53 

Casual labour 6 7 5 9 6 7 16 10 

Regular labour d 87 157 79 49 126 137 69 54 

Equipment 

depreciation 

107 131 80 98 112 119 108 108 

Equipment repairs 41 60 48 79 46 57 60 59 

Fuel and oil 23 28 26 37 25 26 29 23 

Electricity 6 9 8 11 6 7 7 7 

Rent and rates 139 141 130 136 147 158 145 156 

General repairs 11 23 15 11 21 22 15 11 

Miscellaneous 55 61 76 78 56 65 85 94 

Total 498 642 508 540 564 621 565 574 

Net farm income 273 162 230 355 49 59 35 99


BENEFITS OF INTRODUCING BULBS INTO THE ROTATION 

Many farmers would welcome a new crop to grow, particularly on a contract, 

where, price is guaranteed for produce of acceptable quality. Potato and vegetable 

cropping are, however, becoming increasingly concentrated in the hands of 

specialists who are prepared to invest heavily in crop handling and storage 

equipment. The returns from these crops are also unpredictable, particularly 

if irrigation is not available. Further, in the UK, sugar beet production is 

effectively a closed shop, due to the requirement for quota from the British Sugar 

Corporation. 

Recent changes in the relative profitability of combinable crops and alterations 

to support payments under the CAP have generated additional interest in 

alternative crops. Reform of the CAP has been necessary due to the need to 

compete more effectively in world markets, to reduce the cost of agricultural 

support and to discourage accumulation of large food surpluses produced to the 

detriment of the environment. A major series of negotiations in early 1999 

known as ‘Agenda 2000’ brought agreement to revisions to the CAP which 

progressively reduced commodity support from the year 2000 onwards. As a 

result, EU farmers will be exposed to the volatility of world markets and the 

realities of global competition. 

Potential bulb producers can be grouped into two classes, those who already 

grow bulbs, onions, carrots or potatoes, and those who do not grow any of these 

crops. The reason for this division is the existing investment in equipment 

which, members of the first group are already likely to have undertaken. The 

equipment required for bulb handling, storage, drying and hot-water treatment 

is described below, and such investment would not be undertaken lightly by a 

new grower. Farmers in the first group are likely to possess, or have access to, 

the necessary equipment. They will, therefore, have the higher fixed cost 

structure associated with these crops and, in the case of potato growers, the 

narcissus crop would have the following considerable agronomic and financial 

advantages: 

1 An additional high-value crop to incorporate into the rotation (there should 

be a minimum of 5 years between potato crops). 

2 Labour and machinery requirements dovetail well with the potato enterprise: 

Harvesting of bulbs will be complete, except where early potatoes are grown. 

Irrigation will have some overlap, but is generally earlier and for less time for 

bulbs. 

Bulbs can be in and out of store before potatoes go in. 

3 No major pest or disease problems are common to both crops. 

BUDGET CAPITAL COSTS FOR NARCISSUS PRODUCTION 

The capital costs, at 1999 levels, for the establishment of a 10 ha narcissus bulb enterprise 

are presented in Table 5.5. It is assumed that total bulb yields of 350 tonne


Table 5.5 Capital costs for equipment needed in growing Narcissus bulbs a 

Item Size Cost 

1a Pre-HWT warm store (bulk system) 175 t 

Building 19.2 m × 19.2 m × 5.5 m £54 600 

Tunnel 19.2 m × 1.2 m × 1.5 m £6048 

Floor 19.2 m × 18 m £15 225 

Fans 8.27 m 3 /s £1890 

Heater 110 kW £2730 

Total £80 493 

1b Pre-HWT warm store (box system) 175 t 

Building 21 m × 12 m × 5.5 m £36 750 

Letterbox wall 21 m × 4 m £11 025 

Fans 8.27 m 3 /s £1890 


Total £52 395 

2 HWT system 1.25 t/hr 

Tanks 2 × 4 box (3 tonne) £24 150 

3 Post-HWT cooling 175 t 

Building 10 m × 12 m × 6.5 m £19 950 

Fan top system 2 × 4 box, 0.4 m 3 /s £2625 

Total £22 575 

4 Bed former £4037 

5 Stone clod separator £23 189 

6 Bulb planter 2.5 t/h £5250 

7 Haulm topper £4725 

8 Bulb lifter £38 850 

9 Handling equipment 2.5 t/h 

Hopper, soil remover, riddle, inspection table £23 100 

Sprayer £1890 

Conveyor/elevator £4998 

10a Drying system (bulk) 350 t 

Building 30 m × 22 m × 5.5 m £89 250 

Tunnel 30 m × 1.5 m × 1.2 m £9450 

Floor 30 m × 20.8 m £23 100 

Fans 18.6 m 3 /s £3728 


Total £129 938 

10b Drying system (box) 350 t 

Building 24 m × 20 m × 5.5 m £65 100 

Letter box wall × 2 24 m × 3m× 1 m £9660 

Fans 37.2 m 3 /s £5040 


Total £88 620 

11 Boxes 455 × ‘potato boxes’ £22 932 

Note 

aAll costs (at 1999 levels) are purely budgetary estimates and will vary widely with suppliers and manufacturers. 

will be achieved, with 175 tonne bulbs being retained for re-planting, each year. 

The capital items included are: 

1 A pre-hot-water treatment warm store has been specified, both for bulk 

(‘loose’) handling and for handling bulbs in boxes (bulk bins). This function


could well be performed by the main drying store, and hence this cost could 

be avoided. 

2 Well designed and operated hot-water treatment (HWT) equipment is essential. 

However, costs could be shared by the establishment of a central HWT system, 

used by several growers. 

3 Post-HWT cooling is also essential, as is a well laid out treatment site in order 

to prevent re-contamination of treated stock and to ensure that bulbs are 

cooled rapidly following HWT. The cost of a simple building is included, 

though the holding of treated stocks outdoors is practiced by some growers. 

Holding and cooling treated stocks indoors will require suitable ventilation to 

avoid damage to health from the formaldehyde fumes resulting from HWT. 

The cooling of stocks in drying stores after HWT is not recommended. 

4 Bed former. 

5 Stone and clod separator. 

6 Bulb planter. 

7 Haulm topper. 

8 Bulb lifter. 

9 Handling equipment: pre-storage cleaning and post-storage riddling is taken 

as being relatively simple, with no great degree of grading or any bagging being 

included. 

10 Drying equipment is included both for bulk (‘loose’) handling and for handling 

in boxes. 

11 Boxes for where bulbs are handled in bulk bins rather than in loose bulk. 

Whilst all items likely to be required have been identified and costed, a grower 

already growing root crops, potatoes or onions may well have sufficient equipment 

available for the handling of bulbs. Potato or onion harvesting and post-harvesting 

handling equipment and boxes can all be used, with minor modifications, for the 

bulbs. Onion driers/stores operate on very similar principles to bulb stores and 

could therefore be used for bulbs. Redundant potato stores could be adapted for 

bulb drying and storage. 


The author thanks Mrs. J.E. Hawley (ADAS Consulting Ltd) for the provision of 

business management data and help with the text. 

REFERENCES 

ADAS (1984) Narcissus, planting density for 1 and 2 year down crops. ADAS Research and 

Development Reports. Agriculture Service. Bulbs and Allied Flower Crops 1983. Reference Book 

232 (83). MAFF (Publications), Alnwick, pp. 1–2. 

ADAS (1985a) Narcissus Bulb Production. Booklet 2150. MAFF (Publications), Alnwick. 

ADAS (1985b) Narcissus, planting density for one and two year down crops. ADAS Research 

and Development Reports. Agriculture Service. Bulbs and Allied Flower Crops 1984. Reference 

Book 232 (84). MAFF Publications, Alnwick, pp. 2–4.


ADAS (1987a) Narcissus, a comparison of one year versus two year bulb production 

systems. ADAS Research and Development Summary Reports on Bulbs and Allied Flower Crops 

1987, item 5. 

ADAS (1987b) Narcissus, density of planting and crop duration. ADAS Research and Development 

Summary Reports on Bulbs and Allied Flower Crops 1987, item 6. 

University of Nottingham Rural Business Research Unit (1998) Farming in the East Midlands. 

Financial Results 1997–1998. 47th Annual Report, University of Nottingham.

6 Alkaloids of Narcissus 

Jaume Bastida and Francesc Viladomat 

INTRODUCTION 

Since the isolation of the first Amaryllidaceae alkaloid, lycorine (1), from Narcissus 

pseudonarcissus by Gerrad at the end of the last century, more than 200 species and 

varieties belonging to this plant family have been examined for alkaloids. 

Although this group of alkaloids has been of minor pharmaceutical importance 

until recently, there has been an increased interest due to the possible applications 

of galanthamine (69), an alkaloid isolated exclusively from species of this family. 

There are several reviews of Amaryllidaceae alkaloids (Ghosal et al., 1985b; Martin, 

1987) and, likewise, this topic is regularly reviewed by the journal Natural Products 

Reports of The Royal Society of Chemistry (Lewis, 1998, and previous years). This 

chapter covers phytochemical studies on Narcissus alkaloids up to March 1999. 

There has been considerable taxonomical controversy over which genera belong 

to the Amaryllidaceae. The revisions of Dahlgren’s group (Dahlgren, 1980; Dahlgren 

et al., 1985) have contributed to clarifying this aspect. On the other hand, one 

of the best tools for the classification of several genera and species of this family 

has been the type of alkaloids that are found exclusively in Amaryllidaceae. 

Furthermore, it is unusual to find other types of alkaloids in Amaryllidaceae, but if 

present, they are always accompanied by typical Amaryllidaceae alkaloids. Up to 

now, only three alkaloids isolated from this family do not belong to this specific 

type, but to the mesembrane (Sceletium) type generally found in the Aizoaceae 

family (Jeffs, 1981), and for this reason the Amaryllidaceae alkaloids have a high 

chemotaxonomical value. One of these compounds, mesembrenone (82), was 

isolated from Narcissus pallidulus (Bastida et al., 1989) (Figure 6.1). 

The general characteristics of the Amaryllidaceae alkaloids can be summarised 

as follows: 

1 A fundamental ring system composed of a C 6 –C 1 and an N–C 2 –C 6 building 

block, derived from L-phenylalanine and L-tyrosine, respectively. 

2 They are moderately weak bases (pKa values of 6–9). 

3 Each alkaloid contains only one nitrogen atom which is secondary, tertiary or 

even quaternary, and the carbon content varies from 16 to 20 atoms. 

Most of the Amaryllidaceae alkaloids may be classified into nine principal skeletally 

homogeneous subgroups, although there are several other alkaloids with structures 

derived from these main molecular frameworks. Representative alkaloids from

142 J. Bastida and F. Viladomat 

Figure 6.1 Sceletium type alkaloid mesembrenone (82) isolated from N. pallidulus. 

each of these classes include: norbelladine (83), lycorine (1), homolycorine (23), 

crinine (84), haemanthamine (49), narciclasine (63), tazettine (58), montanine (85) 

and galanthamine (69) (Figure 6.2). With the aim of unifying the numbering system 

of the different types, Ghosal’s proposal (Ghosal et al., 1985b) will be used in 

this chapter for the following reasons: 

1 Numbering in the aromatic ring A is always the same (irrespective of the alkaloid 

skeletal type). 

2 The benzylic position, α with respect to the heteroatom, is always 6. 

3 The vicinal position with respect to the nitrogen in the pyrrolidine ring and 

not involved in the ring fusion is always 12. 

4 Numbering in ring C is always clockwise except for homolycorine type alkaloids 

due to their biosynthetic process. 

NARCISSUS ALKALOIDS AND THEIR OCCURRENCE 

The alkaloids isolated from Narcissus species, classified in relation to the different 

ring types, are shown in Table 6.1(a-h). Table 6.2 lists the different Narcissus 

species and cultivars with their corresponding alkaloids. 

BIOSYNTHETIC PATHWAYS 

Work on the biosynthesis of Amaryllidaceae alkaloids reached a peak in the period 

1960–1976, with a great number of studies related to the subject. However, since then, 

little new work has been produced apart from the isolation of compounds predicted 

as biosynthetic intermediaries of a certain pathway or, more recently, a new biosynthetic 

proposal to obtain galanthamine (69) which differs from the initial one.

Figure 6.2 Narcissus alkaloid types.

Table 6.1 Narcissus alkaloid structures 

Table 6.1a Lycorine type 

RO 

3 

RO 

4 

RO 

1 

H 

Alkaloid name Structure 

R2 

H 

N 

R 1 R 2 R 3 R 4 

1 

2 

3 

lycorine 

poetaminine 

pseudolycorine 

H 

Ac 

H 

OH 

OH 

OH H 

CH2 CH2 Me 

4 1-O-acetylpseudolycorine Ac OH H Me 

5 2-O-acetylpseudolycorine H OAc H Me 

6 9-O-methylpseudolycorine H OH Me Me 

7 galanthine H OMe Me Me 

8 goleptine H OMe Me H 

9 

10 

11 

jonquilline 

caranine 

pluviine 

Ac 

H 

H 

O 

H 

H Me 

CH2 CH2 Me 

12 norpluviine H H Me H 

13 9-O-demethylpluviine H H H Me 

14 1-O-acetyl-9-O-demethylpluviine Ac H H Me 

15 1,9-O-diacetyl-9-O-demethylpluviine Ac H Ac Me 

MeO 

MeO 

HO 

OMe 

OH 

H H 

N 

O 

O 

AcO 

H H 

16 narcissidine 17 nartazine 

OAc 

N 

H

MeO 

MeO 

R1 

N 

R2 

MeO 

MeO 

18 assoanine R 1 = R 2 = H 20 vasconine R 1 = H 

19 oxoassoanine R 1 + R 2 = O 21 tortuosine R 1 = OMe 

MeO 

MeO 

22 roserine 

OMe 

Table 6.1b Homolycorine type 

RO 

1 

RO 

2 

RN 

6 

H 

R3 

H 

N 

+ 

H 

O 

R4 

R5 

R1 

N 

+

Table 6.1b Continued 


R1 R2 R3 R4 R5 R6 23 homolycorine Me Me O H Me 

24 8-O-demethylhomolycorine Me H O H Me 

25 8-O-demethyl-8-O-acetylhomolycorine Me Ac O H Me 

26 9-O-demethylhomolycorine H Me O H Me 

27 

28 

29 

masonine 

normasonine 

9-O-demethyl-2α-hydroxyhomolycorine H 

CH2 CH2 Me 

O 

O 

O 

H 

H 

OH 

Me 

H 

Me 

30 

31 

hippeastrine 

lycorenine Me 

CH2 Me 

O 

OH H 

OH 

H 

Me 

Me 

32 O-methyllycorenine Me Me OMe H H Me 

33 

34 

35 

oduline 

6-O-methyloduline 

2α-hydroxy-6-O-methyloduline 

CH2 CH2 CH2 OH 

OMe 

OMe 

H 

H 

H 

H 

H 

OH 

Me 

Me 

Me 

Me-CHOH-CH2-COO 

MeO 

HO 

MeO 

O 

N + 

O 

O 

MeN 

H 

36 dubiusine 

H 

O 

H 

O 

OAc 

MeN 

Me 

H H 

H 

O 

H 

37 8-O-demethylhomolycorine-N-oxide 38 poetinatine 

O 

H 

O 

H 

O 

H 

OCOEt

Table 6.1c Haemanthamine type 

RO 

3 

RO 

4 

R5 

H 


N 

R6 

R1 

R7 

R2 

R1 R2 R3 R4 R5 R6 R7 39 vittatine 

40 maritidine 

OH 

OH 

H 

H 

CH2 Me Me 

H 

H 

H 

H 

H 

H 

41 8-O-demethylmaritidine OH H Me H H H H 

42 9-O-demethylmaritidine OH H H Me H H H 

43 O-methylmaritidine OMe H Me Me H H H 

44 papyramine OMe H Me Me H OH H 

45 6-epipapyramine OMe H Me Me OH H H 

46 O-methyl-6-epipapyramine OMe H Me Me OMe H H 

47 6α-hydroxy-3-O-methylepimaritidine H OMe Me Me H OH H 

48 6β-hydroxy-3-O-methylepimaritidine H OMe Me Me OH H H 

49 haemanthamine 

50 11-O-acetylhaemanthamine 

51 haemanthidine 

52 6-epihaemanthidine 

53 crinamine 

54 narcidine 

OMe 

OMe 

OMe 

OMe 

H 

OMe 

H 

H 

H 

H 

OMe 

H 

CH2 CH2 CH2 CH2 CH2 Me H 

H 

H 

H 

OH 

H 

H 

H 

H 

OH 

H 

H 

H 

OH 

OAc 

OH 

OH 

OH 

OH 

RO 

1 

RO 

2 

R3 

R4 

O 

MeCOOCH2 

H H 

N 

O 

N 

55 cantabricine R1 = H, R2 = Me, 57 bujeine 

R3 = H, R4 = OAc, R5 = H 

56 narcimarkine R1 + R2 = CH2 , 

R3 = OMe, R4 = H, R5 = OCH2CH(OH)Et R5 

O 

OMe

Table 6.1d Tazettine type 

O 

O 

R1 

R2 

O 

H H 

NMe 

NMe 

OH 

58 tazettine R 1 = OMe, R 2 = H 

59 criwelline R 1 = H, R 2 = OMe 60 pretazettine 

61 3-epimacronine 62 obesine 

Table 6.1e Narciclasine and montanine types 

O 

O 

O 

O 

OH 

O 

OMe 

O 

OH 

H 

NH 

O 

H 

NMe 

H 

OH 

OH 

O 

O 

O 

O 

OH 

OMe 

63 narciclasine 64 narciprimine 

O 

O 

OH 

O 

O 

OH 

O 

NH 

H 

NH 

OH

O 

O 

O 

O 

N 

65 trisphaeridine 66 bicolorine 

OH 

NHMe 

67 ismine 68 pancracine 

Table 6.1f Galanthamine type 

MeO 

O 


N 

R1 R2 R3 69 galanthamine OH H Me 

70 epigalanthamine H OH Me 

71 O-acetylgalanthamine OAc H Me 

72 norgalanthamine OH H H 

73 epinorgalanthamine H OH H 

74 N-formylnorgalanthamine OH H CHO 

75 narcisine OH H Ac 

76 narwedine O Me 

R1 

R3 

R2 

O 

O 

O 

O 

N 

N 

+ 

H 

Me 

OH 

OH

Table 6.1f Continued 

MeO 


O 

R1 R2 R3 77 lycoramine OH H Me 

78 norlycoramine OH H H 

79 epinorlycoramine H OH H 

Table 6.1g Alkaloids without phenol oxidative coupling 

MeO 

HO 

HO 

80 O-methylnorbelladine 

Table 6.1h Miscellaneous 

O 

O 

HO 

OMe 

N 

N 

NH 

O 

R1 

R3 

R2 

H 

NMe 

O 

OMe 

N 

Me 

OMe 

H 

OMe 

81 pallidiflorine 82 mesembrenone 

(heterodimer) (Sceletium) 

O

Table 6.2 Occurrence of Narcissus alkaloids 

Species* Alkaloids References 

N. angustifolius Curtis ex Haw. 

= N. poeticus L. subsp. 

radiiflorus (Salisb.) Baker 

N. assoanus Léon-Duf. 18 assoanine 

19 oxoassoanine 

3 pseudolycorine 

4 1-O-acetylpseudolycorine 


N. asturiensis (Jordan) Pugsley 49 haemanthamine 


67 ismine 

61 3-epimacronine 

58 tazettine 

65 trisphaeridine 

N. aureus Loisel. = N. tazetta 

L. subsp. aureus (Loisel.) Baker 

N. bicolor L. 66 bicolorine 

26 9-O-demethylhomolycorine 

67 ismine 


60 pretazettine 

20 vasconine 

N. biflorus Curtis = N. × 

medioluteus Mill. 

N. bujei (Fdez. Casas) Fdez. Casas 57 bujeine 

53 crinamine 



23 homolycorine 


31 lycorenine 

32 O-methyllycorenine 

27 masonine 

34 6-O-methyloduline 

58 tazettine 

N. canaliculatus Guss. 49 haemanthamine 

63 narciclasine 

58 tazettine 

N. cantabricus DC. 55 cantabricine 

53 crinamine 

58 tazettine 

39 vittatine 

N. confusus Pugsley 69 galanthamine 

74 N-formylnorgalanthamine 





N. cyclamineus DC. 63 narciclasine 

76 narwedine 

N. cyclamineus DC. cv. Beryl 7 galanthine 

1 lycorine 

16 narcissidine 

69 galanthamine Cherkasov et al., 1988 

Llabrés et al., 1986a 

Llabrés et al., 1986b 

Viladomat et al., 1997 

1 lycorine Hung et al., 1962 


1 lycorine Hung et al., 1962 

Labraña et al., 1999 

Boit and Döpke, 1956 

Piozzi et al., 1969 

Bastida et al., 1995b 

Bastida et al., 1987a 


Bazhenova et al., 1971 


Boit et al., 1957b

Table 6.2 Continued 


N. cyclamineus DC. cv. Cairhays 7 galanthine 



11 pluviine 

N. cyclamineus DC. cv. 

February Gold 

N. cyclamineus DC. cv. 

Peeping Tom 


77 lycoramine 

31 lycorenine 

31 lycorenine 

11 pluviine 

58 tazettine 

N. dubius Gouan 36 dubiusine 

29 9-O-demethyl-2αhydroxyhomolycorine 


N. eugeniae Fdez. Casas 69 galanthamine 


31 lycorenine 

N. × gracilis Sabine 69 galanthamine 

1 lycorine 

58 tazettine 

N. × incomparabilis Mill. cv. 

Carabiniere 


Daisy Schäffer 


Deanna Durbin 


Flower Record 


Fortune 


Helios 


John Evelyn 

Boit et al., 1957b 




Bastida et al., 1992c 

Codina et al., 1988 

Codina et al., 1992a 


63 narciclasine Piozzi et al., 1969 

69 galanthamine 

7 galanthine 

11 pluviine 

7 galanthine 


1 lycorine 


11 pluviine 


7 galanthine 


1 lycorine 

11 pluviine 



30 hippeastrine 

33 oduline 


7 galanthine 



31 lycorenine 


11 pluviine 


7 galanthine 


1 lycorine 

11 pluviine 

Boit et al., 1957c 

Boit and Ehmke, 

1956 




Gorbunova et al., 1984 


Piozzi et al., 1968, 

1969 

Boit et al., 1957c




Marion Cran 


Mercato 


R.O. Backhouse 


Nova Scotia 


Oranje Bruid 


Pluvius 



7 galanthine 


1 lycorine 





7 galanthine 


1 lycorine 



7 galanthine 


1 lycorine 


11 pluviine 


1956 


N. × incomparabilis Mill. cv. 63 narciclasine Ceriotti et al., 1967 

Scarlet Elegance 


N. × incomparabilis Mill. cv. 7 galanthine 


Sempre Avanti 


Ceriotti et al., 1967 

1 lycorine 




11 pluviine 

1969 

N. × incomparabilis Mill. cv. Suda 69 galanthamine 

7 galanthine 


31 lycorenine 

11 pluviine 


N. × incomparabilis Mill. cv. 7 galanthine 


Toronto 


1 lycorine 

N. × incomparabilis Mill. cv. Tunis 64 narciclasine Ceriotti et al., 1967 


N. × incomparabilis Mill. cv. 64 narciclasine Ceriotti et al., 1967 

Walt Disney 


N. jacetanus Fdez. Casas 18 assoanine 

19 oxoassoanine 

1 lycorine 



N. jonquilla L 69 galanthamine 

77 lycoramine 


Vigneau et al., 1984 

N. jonquilla L. cv. Golden Sceptre 69 galanthamine 

7 galanthine 

8 goleptine 


Döpke, 1963b 

Döpke, 1963c



N. jonquilla L. cv. Golden Sceptre 

(contd.) 




9 jonquilline 

31 lycorenine 

1 lycorine 

27 masonine 

33 oduline 

58 tazettine 

N. jonquilla L. cv. Trevithian 69 galanthamine 

31 lycorenine 

1 lycorine 


58 tazettine 

N. leonensis Pugsley 72 norgalanthamine 

73 epinorgalanthamine 

79 epinorlycoramine 

1 lycorine 

N. lobularis Hort. 69 galanthamine 


N. muñozii-garmendiae Fdez. 

Casas 


31 lycorenine 


N. nivalis Graells 69 galanthamine 

72 norgalanthamine 

6 9-O-methylpseudolycorine 

N. obesus Salisb 66 bicolorine 



67 ismine 


62 obesine 


N. × odorus L. var. rugulosus 69 galanthamine 



1 lycorine 


33 oduline 

58 tazettine 

N. pallidiflorus Pugsley 49 haemanthamine 



67 ismine 

81 pallidiflorine 


N. pallidulus Graells 82 mesembrenone 

22 roserine 

N. panizzianus Parl. 7 galanthine 


Döpke, 1964 

Döpke and Dalmer, 

1965a 



Bastida et al., 1993 










Bastida et al., 1990a



44 papyramine 

45 6-epipapyramine 


N. papyraceus Ker. Gawl. 69 galanthamine 



37 8-O-demethylhomolycorine- 

N-oxide 

77 lycoramine 

1 lycorine 

40 maritidine 

43 O-methylmaritidine 

56 narcimarkine 

44 papyramine 


46 O-methyl-6-epipapyramine 


58 tazettine 

N. poeticus L. 69 galanthamine 

7 galanthine 


31 lycorenine 

1 lycorine 


17 nartazine 

68 pancracine 

2 poetaminine 

11 pluviine 


N. poeticus L. var. ornatus Hort. 69 galanthamine 

7 galanthine 



31 lycorenine 

1 lycorine 

2 poetaminine 

38 poetinatine 

58 tazettine 

N. poeticus L. cv. Actaea 69 galanthamine 

7 galanthine 

31 lycorenine 

1 lycorine 



N. poeticus L. cv. Daphne 69 galanthamine 


31 lycorenine 

1 lycorine 

N. poeticus L. cv. Sarchedon 69 galanthamine 

7 galanthine 

31 lycorenine 

1 lycorine 


Döpke and Sewerin, 

1981 

Hung et al., 1981 

Suau et al., 1990a 


Boit and Stender, 

1954 

Döpke and Sewerin, 

1981 

Harken et al., 1976 

Wildman and Brown, 

1968 

Willaman and Li, 

1970 

Boit, 1954 


Döpke, 1963a 

Döpke and Nguyen, 

1974 




Boit and Döpke, 1956



N. primigenius (Laínz) Fdez. 

Casas and Laínz 




41 8-O-demethylmaritidine 

N. pseudonarcissus L. 80 O-methylnorbelladine 



1 lycorine 

27 masonine 

N. pseudonarcissus L. cv. Carlton 69 galanthamine 

71 O-acetylgalanthamine 





77 lycoramine 

79 epinorlycoramine 

31 lycorenine 


27 masonine 

28 normasonine 


76 narwedine 

33 oduline 

34 6-O-methyloduline 

13 9-O-demethylpluviine 

14 1-O-acetyl- 

9-O-demethylpluviine 

15 1,9-O-diacetyl- 

9-O-demethylpluviine 

39 vittatine 

N. pseudonarcissus L. cv. 

Covent Garden 


Early Glory 


Flower Carpet 


Golden Harvest 


Grand Maître 



7 galanthine 


78 norlycoramine 

31 lycorenine 

1 lycorine 

11 pluviine 


7 galanthine 


1 lycorine 


Cook et al., 1954 

Gude et al., 1988 


1970 

Kreh and Matusch, 

1995 

Kreh et al., 1995a 

Kreh et al., 1995b 



1956 






7 galanthine 



Boit et al., 1957c




Imperator 


King Alfred 

1 lycorine 

58 tazettine 


7 galanthine 



31 lycorenine 

1 lycorine 

11 pluviine 

18 assoanine 



7 galanthine 






31 lycorenine 

1 lycorine 


54 narcidine 


33 oduline 

11 pluviine 


N. pseudonarcissus L. cv. Magnet 7 galanthine 


11 pluviine 


Magnificence 


Mount Hood 


Mrs. Ernst H. Krelage 


Music Hall 



7 galanthine 



1 lycorine 

11 pluviine 




77 lycoramine 




7 galanthine 


1 lycorine 


11 pluviine 

7 galanthine 


1 lycorine 



1956 


Fales et al., 1956 

Fuganti et al., 1974b 

Harken et al., 1976 


1969 

Tojo, 1991 



1956 


Bastos et al., 1996 


Moraes-Cerdeira 

et al., 1997b 




Boit et al., 1957c




Oliver Cromwell 


President Lebrun 


Queen of Bicolors 


Rembrandt 


Rockery Beauty 


7 galanthine 


11 pluviine 


63 narciclasine Ceriotti et al., 1967 


7 galanthine 


1 lycorine 

11 pluviine 





1 lycorine 



7 galanthine 




N. pseudonarcissus L. cv. Romaine 7 galanthine 


31 lycorenine 

11 pluviine 


Spring Glory 


Unsurpassable 


7 galanthine 



1 lycorine 

11 pluviine 



31 lycorenine 

11 pluviine 

N. pseudonarcissus L. cv. Victoria 72 norgalanthamine 

7 galanthine 



1 lycorine 

11 pluviine 

N. pseudonarcissus L. cv. Wrestler 69 galanthamine 

7 galanthine 


11 pluviine 

N. radinganorum Fdez. Casas 23 homolycorine 













N. serotinus Löfl. ex L. 63 narciclasine Piozzi et al., 1969



N. tazetta L. 59 criwelline 


7 galanthine 







1 lycorine 


27 masonine 


75 narcisine 


17 nartazine 

76 narwedine 



58 tazettine 

N. tazetta L. var. chinensis Roem 69 galanthamine 

70 epigalanthamine 




77 lycoramine 

31 lycorenine 

1 lycorine 

40 maritidine 


47 6α-hydroxy-3-O-methylepimaritidine 

48 6β-hydroxy-3-O- methylepimaritidine 

44 papyramine 


11 pluviine 



58 tazettine 

N. tazetta L. cv. Cragford 49 haemanthamine 


1 lycorine 

58 tazettine 

N. tazetta L. cv. Early Perfection 49 haemanthamine 



1 lycorine 

11 pluviine 

58 tazettine 

N. tazetta L. cv. Geranium 69 galanthamine 


Abdallah, 1993 

Abduazimov and 

Yunusov, 1967 

Abou-Donia et al., 

1989 

Bi et al., 1998 



Evidente, 1991 

Evidente et al., 1994 

Furusawa et al., 1976a 


1969 

Späth and Kahovec, 

1934 

Späth et al., 1936 

Tani et al., 1981 


Laing and Clark, 

1974 

Ma et al., 1986 




Ceriotti et al., 1967



N. tazetta L. cv. Geranium 

(contd.) 


1 lycorine 



58 tazettine 

N. tazetta L. cv. La Fiancée 7 galanthine 



1 lycorine 

58 tazettine 

N. tazetta L. cv. Laurens Koster 49 haemanthamine 


1 lycorine 

58 tazettine 

N. tazetta L. cv. L’Innocence 49 haemanthamine 


1 lycorine 


58 tazettine 

N. tazetta L. cv. Scarlet Gem 7 galanthine 



1 lycorine 

58 tazettine 

N. tazetta L. cv. St. Agnes 49 haemanthamine 


1 lycorine 

58 tazettine 

N. tortifolius Fdez. Casas 36 dubiusine 




29 9-O-demethyl-2αhydroxyhomolycorine 

N. tortuosus Haworth 1 lycorine 

21 tortuosine 

N. triandrus L. cv. Silver Chimes 49 haemanthamine 

1 lycorine 

58 tazettine 

N. triandrus L. cv. Thalia 49 haemanthamine 


31 lycorenine 

1 lycorine 


N. triandrus L. cv. Tresamble 69 galanthamine 


31 lycorenine 


N. vasconicus Fdez. Casas 23 homolycorine 

25 8-O-demethyl-8- 

O-acetylhomolycorine 



et al., 1997b 











1969 



1969 

Bastida et al., 1992a



Narcissus L. cv. Celebrity 

1 lycorine 

20 vasconine 


Narcissus L. cv. Cheerfulness 69 galanthamine 



et al., 1997b 


Narcissus L. cv. Clamor 63 narciclasine Ceriotti et al., 1967 


Nancissus L. cv. Folly 1 lycorine 


58 tazettine 

Yunusov, 1967 

Narcissus L. cv. Ice Follies 10 caranine 



et al., 1997a 




77 lycoramine 


1 lycorine 

et al., 1997b 

Narcissus L. cv. Inglescombe 69 galanthamine 

Bastos et al., 1996 




77 lycoramine 


31 lycorenine 

1 lycorine 

11 pluviine 


Narcissus L. cv. Insulinde 7 galanthine 


1 lycorine 

11 pluviine 


Narcissus L. cv. Irene Copeland 1 lycorine 

76 narwedine 


Narcissus L. cv Kristalli 69 galanthamine 


1 lycorine 

76 narwedine 

58 tazettine 

Yunusov, 1967 

Narcissus L. cv. Livia 10 caranine 

7 galanthine 


1 lycorine 

11 pluviine 


Narcissus L. cv. Salome 53 crinamine 



35 2α-hydroxy-6-Omethyloduline 


21 tortuosine 

20 vasconine 

Almanza et al., 1996 

Narcissus L. cv. Texas 69 galanthamine 

Barton and Kirby, 1962 


Boit et al., 1957c




Narcissus L. cv. Texas 

(contd.) 

Narcissus L. cv. Totus 

Albus Prior = N. papyraceus 

Ker. Gawl. 

1 lycorine 


76 narwedine 

11 pluviine 

12 norpluviine 

Narcissus L. cv. Twink 69 galanthamine 

7 galanthine 


1 lycorine 

11 pluviine 

Narcissus L. cv. Van Sion 69 galanthamine 


7 galanthine 




31 lycorenine 

1 lycorine 

11 pluviine 

58 tazettine 


Kirby and Michael, 

1973 

Kirby and Tiwari, 

1966 


63 narciclasine Ceriotti et al., 1967 



1956 



1956 


Narcissus L. cv. Verger 63 narciclasine Ceriotti et al., 1967 


Narcissus sp. 69 galanthamine 


64 narciprimine 

2 poetaminine 

Ceriotti, 1967 

Cherkasov et al., 1989 



1970 

Note 

*The taxonomical aspects are based on the works of Barra and López-González (1984a,b), Blanchard 

(1990), Dorda and Fernández-Casas (1984a,b, 1989, 1990, 1994), Dorda et al. (1991), Fernandes (1975, 

1991), Fernández-Casas (1983, 1984a,b, 1986) and Pugsley (1933) for wild Narcissus species and 

hybrids, and those of Boit and Döpke (1956), Boit and Ehmke (1956), Boit et al. (1957b,c), Jefferson- 

Brown (1991), Kington (1989) and Piozzi et al. (1969) for Narcissus cultivars. 

Although L-phenylalanine (L-phe) and L-tyrosine (L-tyr) are closely related in 

chemical structure, they are not interchangeable in plants. In the Amaryllidaceae 

alkaloids, L-phe serves as a primary precursor of the C 6 –C 1 fragment, corresponding 

to ring A and the benzylic position (C-6), and L-tyr is the precursor of ring C, 

the two-carbon side chain (C-11 and C-12) and nitrogen, C 6 –C 2 –N. The conversion 

of L-phe to the C 6 –C 1 unit requires the loss of two carbon atoms from the side chain 

as well as the introduction of at least two oxygenated substituents into the aromatic 

ring, which is performed via cinnamic acids. The presence of the enzyme phenylalanine 

ammonia lyase (PAL) has been demonstrated in Amaryllidaceae (Suhadolnik 

et al., 1963), and the elimination of ammonia mediated by this enzyme is known 

to occur in an antiperiplanar manner to give trans-cinnamic acid, with loss of the

Alkaloids of Narcissus 163 

β-pro-S hydrogen (Wightman et al., 1972). Thus, it may be expected that L-phe 

would be incorporated into Amaryllidaceae alkaloids with retention of the β-pro-R 

hydrogen. However, feeding experiments in Narcissus pseudonarcissus cv. King Alfred 

showed that tritium originally present at C-β of L-phe, whatever the configuration, 

was lost in the formation of several haemanthamine and homolycorine type alkaloids, 

which led to the conclusion that fragmentation of the cinnamic acids involves 

oxidation of C-β to ketone or acid level, the final product being protocatechuic 

aldehyde or its derivatives (Figure 6.3). On the other hand, L-tyr is degraded no 

further than tyramine before incorporation into the Amaryllidaceae alkaloids. 

Thus, tyramine and protocatechuic aldehyde or its derivatives are logical components 

for the biosynthesis of the precursor norbelladine (83). This reaction 

occupies a pivotal position since it represents the entry of primary metabolites into 

a secondary metabolic pathway. The junction of the amine and the aldehyde 

results in a Schiff’s base, two of which have been isolated up to now: craugsodine 

(Ghosal et al., 1986) and isocraugsodine (Ghosal et al., 1988a). The existence of 

Schiff’s bases in nature, as well as their easy conversion into the different ringsystems 

of the Amaryllidaceae alkaloids, allows the presumption that the initial 

postulate about this biosynthetic pathway was correct. 

Barton and Cohen (1957) proposed that norbelladine (83) or related compounds 

could undergo oxidative coupling of phenols in Amaryllidaceae plants, once ring 

A had been suitably protected by methylation, resulting in the different skeletons 

of the Amaryllidaceae alkaloids (Figure 6.4). 

Lycorine and homolycorine types 

The alkaloids of this group are derivatives of the pyrrolo[de]phenanthridine alkaloids 

(lycorine type) and the 2-benzopirano-[3,4-g]indole alkaloids (homolycorine 

type), and both types originate from an ortho-para′ phenol-oxidative coupling 

(Figure 6.5). 

The biological conversion of cinnamic acid via hydroxylated cinnamic acids into 

the C 6 –C 1 unit of norpluviine (12) has been used in a study of hydroxylation 

mechanisms in higher plants (Bowman et al., 1969). When [3- 3 H, β- 14 C]cinnamic 

acid was fed to Narcissus pseudonarcissus cv. Texas, a tritium retention in norpluviine 

(12) of 28% was observed. This is very near a predicted value of 25%, resulting from 

para-hydroxylation with hydrogen migration and retention, where half the tritium 

would be lost in the first hydroxylation and half the remainder in the second. 

In the conversion of O-methylnorbelladine (80) into lycorine (1), the labeling 

position [3- 3 H] on the aromatic ring of L-tyr afterwards appears at C-2 of norpluviine 

(12), which is formed as an intermediate, the configuration of the tritium 

apparently being β (Kirby and Tiwari, 1966). This tritium is retained in subsequently 

formed lycorine (1), which means that hydroxylation at C-2 proceeds with 

an inversion of configuration (Bruce and Kirby, 1968) by a mechanism involving 

an epoxide, with ring opening followed by allylic rearrangement of the resulting 

alcohol (Figure 6.6). Supporting evidence comes from the incorporation of [2β- 

3 H]caranine (10) into lycorine (1) in Zephyranthes candida (Wildman and Heimer, 

1967). However, a hydroxylation of caranine (10) in Clivia miniata occurring 

with retention of configuration was also observed (Fuganti and Mazza, 1972b). 

Further, [2α- 3 H; 11- 14 C]caranine (10) was incorporated into lycorine (1) with high


Figure 6.3 Biosynthetic pathway to norbelladine. 

retention of tritium at C-2, indicating that no 2-oxo-compound can be implicated 

as an intermediate. 

The conversion of the O-methoxyphenol to the methylenedioxy group may 

occur late in the biosynthetic pathway. Tritiated norpluviine (12) is converted to

Figure 6.4 Oxidative phenyl-phenyl coupling in Amaryllidaceae alkaloids. 

Figure 6.5 Alkaloids proceeding from an ortho-para′ coupling.


Figure 6.6 Biosynthesis of lycorine (1) with inversion of the configuration. 

Figure 6.7 Conversion of galanthine (7) to narcissidine (16) via epoxide (86). 

tritiated lycorine (1) by Narcissus × incomparabilis cv. Deanna Durbin, which not only 

demonstrates the previously mentioned conversion but also indicates that the C-2 

hydroxyl group of lycorine (1) is derived by allylic oxidation of either norpluviine 

(12) or caranine (10) (Battersby et al., 1964). 

Regarding the conversion of [2β- 3 H, 8-OMe- 14 C]pluviine (11) into galanthine 

(7), in Narcissus pseudonarcissus cv. King Alfred, the retention of 79% of the tritium 

label confirms that hydroxylation of C-2 may occur with inversion of configuration 

(Harken et al., 1976). 

It was considered (Fuganti et al., 1974a) that another analogous epoxide 86 

could give narcissidine (16) in the way shown by loss of the pro-S hydrogen from 

C-11, galanthine (7) being a suitable substrate for epoxidation. Labelled [α- 14 C, 

β- 3 H]-O-methylnorbelladine (80), when fed to Narcissus x incomparabilis cv. Sempre 

Avanti, afforded galanthine (7) (98% of tritium retention) and narcissidine (16) 

(46% tritium retention). Loss of hydrogen from C-11 of galanthine (7) was therefore 

stereospecific. In this decade, Kihara et al. (1992) have isolated a new alkaloid, 

incartine (86), from flowers of Lycoris incarnata, which could be considered to be 

the biosynthetic intermediate of this pathway (Figure 6.7). 

The biological conversion of protocatechuic aldehyde into lycorenine (31), 

which proceeds via O-methylnorbelladine (80) and norpluviine (12), first involves 

a reduction of the aldehyde carbonyl, and afterwards, in the generation of lycorenine 

(31), oxidation of this same carbon atom. The absolute stereochemistry of 

these processes has been elucidated in subsequent experiments (Fuganti and 

Mazza, 1973), and the results show that hydrogen addition and removal take place 

on the re-face of the molecules concerned (Hanson, 1966), the hydrogen initially 

introduced being the one later removed (Fuganti and Mazza, 1971a). It is noteworthy 

that norpluviine (12), unlike pluviine (11), is converted in Narcissus pseudonarcissus 

cv. King Alfred primarily to alkaloids of the homolycorine type. Benzylic oxidation


Figure 6.8 Conversion of norpluviine (12) to homolycorine type alkaloids. 

of position 6 would afford 87, followed by ring opening to form an amino aldehyde, 

and then a hemiacetal formation and methylation could provide lycorenine 

(31) (Harken et al., 1976), and, on subsequent oxidation, could give homolycorine 

(23) as can be seen in Figure 6.8. 

Crinine, haemanthamine, tazettine, narciclasine and montanine types 

This group includes the alkaloids derived from 5,10b-ethanophenanthridine (crinine 

and haemanthamine types), 2-benzopyrano[3,4-c]indole (tazettine type), phenanthridine 

(narciclasine type) and 5,11-methanomorphanthridine (montanine type), 

originating from a para-para′ oxidative phenolic coupling (Figure 6.9). 

Results of experiments with labelled crinine (84), and less conclusively with 

oxovittatine, indicate that the two naturally occurring enantiomeric series, represented 

in Figure 6.9 by crinine (84) and vittatine (39), are not interconvertible in 

Nerine bowdenii (Feinstein and Wildman, 1976). 

Incorporation of O-methylnorbelladine (80), labelled in the methoxy carbon 

and also in positions [3,5- 3 H], into the alkaloid haemanthamine (49), was without 

loss of tritium, half of which was sited at C-2 of (49). Consideration of the possible 

mechanisms involved in relation to tritium retention led to the suggestion that the 

tritium which is expected at C-4 of (49) might not be stereospecific (Fuganti, 1969). 

The conversion of O-methylnorbelladine (80) into haemanthamine (49) involves loss 

of the pro-R hydrogen from the C-β of the tyramine moiety, as well as a further 

entry of a hydroxyl group at this site (Battersby et al., 1971). The subsequent benzylic 

oxidation results in an epimeric mixture 51/52, which even High Performance 

Liquid Chromatography (HPLC) cannot separate. The epimeric forms were proposed 

to be interconvertible through 88a. The biosynthetic conversion of the 5,10b-ethanophenanthridine 

alkaloids to the 2-benzopyrano[3,4-c]indole was demonstrated 

by feeding tritium-labelled alkaloids to Sprekelia formosissima. It was shown that


Figure 6.9 Alkaloids proceeding from a para-para′ coupling. 

this plant converts haemanthamine (49) to haemanthidine/epihaemanthidine (51/52) 

and subsequently to pretazettine (60) in an essentially irreversible manner (Fales 

and Wildman, 1964). This transformation was considered to proceed through 88a 

or the related alkoxide anion, although this intermediate 88a and its rotational 

equivalent 88b have never been detected by spectral methods (Wildman and 

Bailey, 1969) (Figure 6.10). 

It has also been proved that the alkaloid narciclasine (63) proceeds from the 

pathway of the biosynthesis of crinine and haemanthamine type alkaloids and not 

through norpluviine (12) and lycorine (1) derivatives. In fact, in view of its structural 

affinity to both haemanthamine (49) and lycorine (1), narciclasine (63) could be 

derived by either pathway. When O-methylnorbelladine (80), labelled in the methoxy 

carbon and in both protons of position 3 and 5 of the tyramine aromatic ring, was 

administered to narcissus plants, all four alkaloids incorporated activity. The isotopic 

ratio [ 3 H: 14 C] for norpluviine (12) and lycorine (1) was, as expected, 50% that of the 

precursor, because of its ortho-para′ coupling. On the contrary, in haemanthamine 

(49) the ratio was unchanged. These results prove that the methoxy group of (80) 

is completely retained in the alkaloids mentioned, providing a satisfactory internal 

standard, and also the degree of tritium retention is a reliable guide to the direction 

of phenol coupling. Narciclasine (63) showed an isotopic ratio (75%) higher than 

that of lycorine or norpluviine (12) though lower than that of hemanthamine (49). 

However, the fact that more than 50% of tritium is retained suggests that O-methylnorbelladine 

(80) is incorporated into narciclasine (63) via para-para′ phenol 

oxidative coupling. 

O-methylnorbelladine (80) and vittatine (39) are implicated as intermediates in 

the biosynthesis of narciclasine (63) (Fuganti et al., 1971; Fuganti and Mazza,

Figure 6.10 Biosynthesis of pretazettine (60). 


1971b, 1972a), and the loss of the ethane bridge from the latter could occur by a 

retro-Prins reaction on 11-hydroxyvittatine (89). Strong support for this pathway 

was obtained by labeling studies. 11-Hydroxyvittatine (89) has also been proposed 

as an intermediate in the biosynthesis of haemanthamine (49) and montanine (85) 

(a 5,11-methanomorphanthridine alkaloid) following the observed specific incorporation 

of vittatine (39) into the two alkaloids in Rhodophiala bifida (Feinstein and 

Wildman, 1976) (Figure 6.11). 

Fuganti and Mazza (1971b, 1972a) concluded that in the late stages of narciclasine 

(63) biosynthesis, the two-carbon bridge is lost from the oxocrinine skeleton, passing 

through intermediates bearing a pseudoaxial hydroxy-group at C-3 position and 

further hydrogen removal from this position does not occur. Noroxomaritidine was 

also implicated in the biosynthesis of narciclasine (63), and further experiments 

(Fuganti, 1973) showed that it is also a precursor for ismine (67). 

The alkaloid ismine (67) has also been shown (Fuganti and Mazza, 1970) to be a 

transformation product of the crinine-haemanthamine series. The precursor, oxocrinine 

labelled with tritium in the positions 2 and 4, was administered to Sprekelia 

formosissima plants and the radioactive ismine (67) isolated was shown to be specifically 

labelled at the expected positions. 

Galanthamine type 

The alkaloids with a dibenzofuran nucleus (galanthamine type) are obtained from 

a para-ortho′ phenyl oxidative coupling. 

Although norbelladine (83) was shown not to be a precursor of galanthamine 

(69) in Narcissus pseudonarcissus cv. King Alfred, incorporation of this labelled 

compound has been obtained in Leucojum aestivum (Fuganti, 1969). 

The initial studies of this pathway suggested that the phenyl oxidative coupling 

does not proceed from O-methylnorbelladine (80) but that the order of methyla-


Figure 6.11 Proposed biosynthetic pathways to haemanthamine (49) and montanine (85). 

tion of the precursors should be norbelladine (83) --> N-methylnorbelladine --> 

N,O-dimethylnorbelladine (90) to give finally galanthamine (69) (Barton et al., 

1963), the conversion of 91 to narwedine (76) either being not reversible or, if so, 

enzymically controlled (Fuganti, 1969). The precursor N,O-dimethylnorbelladine 

(90) was first isolated in 1988 from Pancratium maritimum (Vázquez-Tato et al., 

1988), a species that also contains galanthamine (69) (Figure 6.12a). 

Chlidanthine (92), by analogy with the known conversion of codeine to morphine, 

might be expected to arise from galanthamine (69) by O-demethylation. This was 

shown to be true when both galanthamine (69) and narwedine (76), with tritium 

labels, were incorporated into chlidanthine (92) (Bhandarkar and Kirby, 1970). 

However, the most recent study seems to contradict the evidence set forth here. 

Experiments carried out with application of 13 C-labelled O-methylnorbelladine 

(80) to organs of field grown Leucojum aestivum have shown that the biosynthesis of 

galanthamine (69) involves the phenol oxidative coupling of O-methylnorbelladine 

(80) to a postulated dienone which undergoes spontaneous closure of the ether 

bridge to yield N-demethylnarwedine (93), giving norgalanthamine (72) after 

stereoselective reduction. Furthermore, it was shown that norgalanthamine (72) is


Figure 6.12a Biosynthesis of galanthamine (69) proposed by Barton et al. (1963). 

Figure 6.12b Biosynthesis of galanthamine (69) proposed by Eichhorn et al. (1998). 

N-methylated to galanthamine (69) in the final step of biosynthesis (Eichhorn et al., 

1998) (Figure 6.12b). In contrast with the literature, N,O-dimethylnorbelladine 

(90) was metabolised to a lesser extent in L. aestivum and incorporated into 

galanthamine (69) as well as norgalanthamine (72) at about one-third the rate of 

O-methylnorbelladine (80). 

According to Eichhorn et al. (1998), narwedine (76) is not the direct precursor of 

galanthamine (69), and could possibly exist in an equilibrium with galanthamine 

(69), a reaction catalysed by a hypothetically reversible oxido-reductase. 

SPECTROSCOPY 

Only Proton Nuclear Magnetic Resonance ( 1 H-NMR), Carbon 13 Nuclear Magnetic 

Resonance ( 13 C-NMR) and Mass Spectrometry (MS), the three most important 

spectroscopic methods for the Amaryllidaceae alkaloids, will be treated here. A list 

of the different narcissus alkaloids and their spectroscopic properties is given in 

Table 6.3. The literature with the most recent spectroscopic data for these alkaloids 

is given, even when they were isolated from species other than Narcissus. 

Table 6.3 Narcissus alkaloids – spectroscopic data 

Alkaloid* Formula MW Spectroscopic data References 

18 assoanine C17H17NO2 267 UV, IR, MS, 

1 13 

H NMR, C NMR 

19 oxoassoanine C17H15NO3 281 UV, IR, MS, 

1 13 

H NMR, C NMR 

66 bicolorine C15H12NO2 238 IR, MS, 1 H NMR, 

13 

C NMR 

57 bujeine C20H23NO6 373 IR, MS, 1 H NMR, 

13 

C NMR, CD 




Labraña et al., 1999



55 cantabricine C18H23NO4 317 IR, MS, 1 H NMR, 

13 

C NMR 


10 caranine C16H17NO3 271 UV, IR, MS, 

1 

H NMR 


CD 

Kuriyama et al., 1967 

53 crinamine C17H19NO4 301 UV, 1 H NMR Likhitwitayawuid et al., 

1993 

IR, MS, CD Viladomat et al., 1994 

13 

C NMR 


X-Ray 

Roques et al., 1977 

59 criwelline C18H21NO5 331 UV 

Hauth and Stauffacher, 

1964 

MS 

Duffield et al., 1965 

1 13 

H NMR, C NMR Razafimbello et al., 1996 

CD 

De Angelis and Wildman, 

1969a 

36 dubiusine C23H27NO8 445 UV, IR, MS, 

1 13 

H NMR, C NMR 


69 galanthamine C17H21NO3 287 UV, IR, MS, 

1 

H NMR 


13 

C NMR 

Abdallah et al., 1989 

CD 


1969a 

X-Ray 

Peeters et al., 1997 

71 O-acetylgalanthamine 

C19H23NO4 329 UV, MS, 1 H NMR, 

13 

C NMR 


70 epigalanthamine C17H21NO3 287 UV, MS 

IR, 1 H NMR 

Kametani et al., 1969 

Vlahov et al., 1989 

CD 


1969a 

72 norgalanthamine C16H19NO3 273 UV 

IR, MS, 1 H NMR, 

13 

C NMR 

Laiho and Fales, 1964 


CD 

Li et al., 1987 

X-Ray 

Roques and Lapasset, 

1976 

73 epinorgalanthamine C16H19NO3 273 IR, MS, 1 H NMR, 

13 

C NMR 


74 N-formylnorgalanthamine 

C17H19NO4 301 UV, IR, MS, 

1 13 

H NMR, C NMR 


7 galanthine C18H23NO4 317 UV 

MS, 1 H NMR, 

13 

C NMR 

Kobayashi et al., 1977 


8 goleptine C17H21NO4 303 IR Döpke, 1964 

49 haemanthamine C17H19NO4 301 UV, IR, MS 

1 

H NMR 


Pabuççuoglu et al., 1989 

13 

C NMR, CD Baudouin et al., 1994 

X-Ray 

Watson et al., 1984 


C19H21NO5 343 IR, MS, 1 H NMR, 

13 

C NMR, CD 

Labraña et al., 1999



51 haemanthidine C17H19NO5 317 UV 

IR, MS 

1 

H NMR 

13 

C NMR 

CD 

52 6-epihaemanthidine C17H19NO5 317 UV 

IR, MS 

1 

H NMR 

13 

C NMR 

CD 

30 hippeastrine C17H17NO5 315 UV 


13 

C NMR 

CD 

23 homolycorine C18H21NO4 315 UV, IR, MS, 

1 

H NMR 

13 

C NMR 

CD 


37 8-O-demethylhomolycorine- 

N-oxide 

25 8-O-demethyl-8-Oacetylhomolycorine 


29 9-O-demethyl- 

2α-hydroxyhomolycorine 

C 17H 19NO 4 301 UV, IR, 1 H NMR, 

13 C NMR 

MS 

CD 

X-Ray 

C 17 H 19 NO 5 317 UV, IR, MS, 

1 H NMR, 13 C NMR 

C 19H 21NO 5 343 IR, MS, 1 H NMR, 

13 C NMR 

C 17H 19NO 4 301 UV, IR, MS, 


C 17 H 19 NO 5 317 IR, MS, 1 H NMR, 

13 C NMR 

67 ismine C 15H 15NO 3 257 UV, IR, MS, 

1 H NMR 


X-Ray 

Irie et al., 1959 

Trimiño et al., 1989 


Antoun et al., 1993 

Wagner et al., 1996 










Jeffs et al., 1985 


Latvala et al., 1995b 



Latvala et al., 1995a 





Suau et al., 1990b 



9 jonquilline C18H17NO5 327 UV, IR Döpke and Dalmer, 1965a 

77 lycoramine C17H23NO3 289 IR, MS, 1 H NMR, 

13 

C NMR 

Li et al., 1987 

13 

C NMR 


13 

C NMR 

Youssef and Frahm, 1998 

78 norlycoramine C16H21NO3 275 IR, MS, 1 H NMR Kihara et al., 1987 

79 epinorlycoramine C16H21NO3 275 IR, MS, 1 H NMR, 

13 

C NMR 


31 lycorenine C18H23NO4 317 UV 

MS 

Kitagawa et al., 1955 

Ibuka et al., 1966 

1 13 

H NMR, C NMR Codina et al., 1992a 

X-Ray 

Clardy et al., 1972



32 O-methyllycorenine C19H25NO4 331 IR, MS, 1 H NMR, 

13 

C NMR 

X-Ray 

1 lycorine C16H17NO4 287 UV, IR, MS, 

1 13 

H NMR, C NMR 

13 

C NMR 

CD 

X-Ray 

61 3-epimacronine C18H19NO5 329 IR, MS, 13 C NMR 

1 

H NMR 

CD 

X-Ray 

40 maritidine C17H21NO3 287 UV 

IR 

MS, 1 H NMR 

1 13 

H NMR, C NMR 

CD 


Labraña et al., 1999 

Likhitwitayawuid et al., 

1993 

Spohn et al., 1994 


Gopalakrishna et al., 1976 


Kihara et al., 1987 


X-Ray 

Linden et al., 1998 


Tomioka et al., 1977 

Ghosal et al., 1985a 



1969b 

Zabel et al., 1979 


C16H19NO3 273 IR, MS 

1 

H NMR 



13 

C NMR 



C16H19NO3 273 IR, MS, 1 H NMR Bastida et al., 1988c 

43 O-methylmaritidine C18H23NO3 301 UV, IR, MS, 

1 13 

H NMR, C NMR 


CD 

Ma et al., 1986 

47 6α-hydroxy-3-Omethylepimaritidine 

C18H23NO4 317 UV, IR, MS, 

1 

H NMR, CD 

Ma et al., 1986 

48 6β-hydroxy-3-Omethylepimaritidine 

C18H23NO4 317 UV, IR, MS, 

1 

H NMR, CD 

Ma et al., 1986 

27 masonine C17H17NO4 299 UV, IR, MS, 

1 13 

H NMR, C NMR 

Kreh and Matusch, 1995 

CD 


28 normasonine C16H15NO4 285 UV, IR, MS, 

1 13 

H NMR, C NMR 


82 mesembrenone C17H21NO3 287 IR, MS, 1 H NMR 

13 

C NMR 



63 narciclasine C14H13NO7 307 UV, IR 

MS, 1 H NMR, 

13 

C NMR 



X-Ray 

Bi et al., 1998 

54 narcidine C17H21NO4 303 UV, IR, MS, 

1 

H NMR 

Tojo, 1991 

56 narcimarkine C21H27NO5 373 IR, MS Döpke and Sewerin, 

1981 

64 narciprimine C14H9NO5 271 UV, IR, 1 H NMR 

MS 


Spenglers and Trimiño, 

1989



75 narcisine C18H21NO4 315 UV, IR, MS, 

1 13 

H NMR, C NMR 

16 narcissidine C18H23NO5 333 UV 

IR, X-Ray 

MS 

1 

H NMR 

X-Ray 

Abdallah, 1993 

Fales and Wildman, 1958 

Clardy et al., 1970 

Kinstle et al., 1966 


Immirzi and Fuganti, 

1971 

17 nartazine C 20H 23NO 6 373 IR Boit and Döpke, 1956 

76 narwedine C 17H 19NO 3 285 UV 

IR, MS, 1 H NMR 

80 O-methylnorbelladine 

C 16H 19NO 3 273 IR, MS, 1 H NMR, 

13 C NMR 

62 obesine C16H17NO4 287 MS, 1 H NMR, 

13 

C NMR 

33 oduline C17H19NO4 301 UV, IR, MS, 

1 13 

H NMR, C NMR 

34 6-O-methyloduline C18H21NO4 315 UV, IR, MS, 

1 13 

H NMR, C NMR 

35 2α-hydroxy-6-Omethyloduline 

C 18H 21NO 5 331 IR, MS, 1 H NMR, 

13 C NMR 

81 pallidiflorine C34H40N2O7 588 IR, MS, 1 H NMR, 

13 

C NMR 

68 pancracine C16H17NO4 287 UV, MS, 

1 13 

H, C NMR, CD 

44 papyramine C18H23NO4 317 UV 


13 

C NMR 

45 6-epipapyramine C18H23NO4 317 UV 


13 

C NMR 

46 O-methyl-6epipapyramine 

C 19 H 25 NO 4 331 UV, IR, MS, 


11 pluviine C 17 H 21 NO 3 287 UV 

IR 

13 9-O-demethylpluviine 

14 1-O-acetyl-9-Odemethylpluviine 

15 1,9-O-diacetyl-9-Odemethylpluviine 

C 16 H 19 NO 3 273 UV, MS, 1 H NMR, 

13 C NMR 

C 18H 21NO 4 315 UV, MS, 1 H NMR, 

13 C NMR 

C 20H 23NO 5 357 UV, MS, 1 H NMR, 

13 C NMR 

12 norpluviine C 16H 19NO 3 273 UV 

IR 

Kametani et al., 1969 

Li et al., 1987 

Machocho et al., 1999 






Ali et al., 1984 






Kirby and Tiwari, 1966 

Boit et al., 1957a 




Kirby and Tiwari, 1966 

Sandberg and Michel, 

1968 

2 poetaminine C 18H 19NO 5 329 UV, IR Döpke and Dalmer, 

1965b




38 poetinatine C20H23NO6 373 IR, MS, 1 H NMR Döpke and Nguyen, 1974 

60 pretazettine C18H21NO5 331 UV, IR, MS, 

1 

H NMR 

Ghosal et al., 1984 

CD 


3 pseudolycorine C16H19NO4 289 UV, IR, MS, 

1 13 

H NMR, C NMR 





C 18H 21NO 5 331 UV, IR, MS, 

1 H NMR 

Note 

* Alkaloids are listed in alphabetical order with disruption of the derivatives that follow the principal 

alkaloid. 

Proton nuclear magnetic resonance 

C 18H 21NO 5 331 UV, IR, MS, 


C 17 H 21 NO 4 303 UV, MS, 1 H NMR 

IR 

22 roserine C18H22NO3 300 MS, 1 H NMR, 

13 

C NMR 

58 tazettine C18H21NO5 331 UV 

IR 

MS, 1 21 tortuosine 

H NMR, 

13 

C NMR 

CD 

X-Ray 

C18H18NO3 296 IR, MS, 1 H NMR, 

13 

C NMR 

65 trisphaeridine C14H9NO2 223 UV, IR, MS, 

1 

H NMR 

13 

C NMR 

20 vasconine C17H16NO2 266 IR, MS 

1 13 

H NMR, C NMR 

39 vittatine C 16H 17NO 3 271 UV, IR, MS, 


1 H NMR 

13 C NMR 

CD 




Fales et al., 1956 


Wildman and Kaufman, 

1954 


Ghosal et al., 1984 

Crain et al., 1971 


Ide et al., 1996 


Suau et al., 1990b 




Vázquez-Tato et al., 1988 


Frahm et al., 1985 


1 H-NMR spectroscopy gives important information about the different types 

of Amaryllidaceae alkaloids. Several early contributions about homolycorine and 

crinane-haemanthamine type alkaloids were made by Hawksworth et al. (1965) 

and Haugwitz et al. (1965). In the last decade, the routine use of 2D-NMR techniques 

(Correlated Spectroscopy (COSY), Nuclear Overhauser Enhancement and 

Exchange Spectroscopy (NOESY), Rotating Frame Nuclear Overhauser Effect 

Spectroscopy (ROESY), etc.) has facilitated the structural assignments and the 

settling of their stereochemistry.


Lycorine type 

This group has been subject to several 1 H-NMR studies, and lycorine (1) as well as 

its main derivatives have been completely assigned. The general characteristics of 

the 1 H-NMR spectra are: 

1 Two singlets for the para-oriented aromatic protons in the range δ 6.5–7.2 

ppm. 

2 A unique olefinic proton around 5.5 ppm. 

3 Two doublets as an AB system corresponding to the benzylic protons of C-6. 

4 The deshielding observed in the β protons of positions 6 and 12 in relation to 

their α-homologs is due to the effect of the cis-lone pair of the nitrogen atom. 

5 Like almost all other lycorine type examples, the alkaloids isolated from 

narcissus show a trans B/C ring junction, the coupling constant being 

J 4a,10b ~11Hz. 

In the plant, the alkaloid lycorine (1) is particularly vulnerable to the oxidation 

processes, giving several ring-C aromatised products. 

Homolycorine type 

This group includes lactone, hemiacetal or cyclic ether alkaloids. The general 

traits for these types of compounds could be summarised as follows: 

1 Two singlets for the para-oriented aromatic protons. In lactone alkaloids, 

differentiation of the H-7 and H-10 signals is readily made by virtue of the 

deshielding of H-7 by effect of the peri-carbonyl group. 

2 The hemiacetal alkaloids always show the substituent at C-6 in α-disposition, and 

the benzylic proton H-6β appears as a singlet between 5–6 ppm, depending on 

the substituent at C-6. 

3 The majority of compounds belong to a single enantiomeric series containing 

a cis B/C ring junction, which is made clear by the small size of the coupling 

constant J 1,10b . In the Narcissus genus no exception to this rule has been 

observed. 

4 The large coupling constant between H-4a and H-10b (J 4a,10b ~10Hz), is only 

consistent with a trans-diaxial relationship. 

5 In general, the C ring presents a vinylic proton around 5.5 ppm. 

6 The singlet corresponding to the N-methyl group is in the range δ 2.0–2.2 

ppm, its non-existence being very unusual. 

7 If position 2 is substituted by an OH, OMe or OAc group, it always displays an 

α-disposition. 

8 The H-12α is more deshielded than H-12β as a consequence of the cis-lone 

pair of the nitrogen atom. 

An interesting study of homolycorine type alkaloids with a saturated ring C 

has been made by Jeffs et al. (1988). They describe empirical correlations of 

N-methyl chemical shifts with stereochemical assignments of the B/C and C/D 

ring junction.


Crinine – haemanthamine types 

The absolute configuration of these alkaloids, which allows both series to be 

differentiated, is determined through the circular dichroism spectrum. The 

alkaloids of narcissus belong to the haemanthamine type, while in genera such as 

Brunsvigia, Boophane, etc., the crinine type alkaloids are predominant. It is also 

noteworthy that the alkaloids isolated from narcissus do not show additional 

substitutions in the aromatic ring apart from those of C-8 and C-9. On the 

contrary, in the genera where crinine type alkaloids predominate, the presence 

of compounds with a methoxy substituent at C-7 is quite common. Thus, taking 

into account the previous considerations, haemanthamine type alkaloids show 

the following characteristics: 

1 Two singlets for the para-oriented aromatic protons in the range δ 6.4–7.0 

ppm. 

2 Using CDCl 3 as the solvent, the magnitude of the coupling constants between 

each olefinic proton (H-1 and H-2) and H-3 gives information about the 

configuration of the C-3 substituent. Thus, in those alkaloids in which the twocarbon 

bridge (C-11 and C-12) was cis to the substituent at C-3, H-1 shows an 

allylic coupling with H-3 ( J 1,3 ~1–2 Hz) and H-2 a smaller coupling with H-3 

( J 2,3 ~0–1.5 Hz), as occurs in crinamine (53). On the contrary, in the corresponding 

C-3 epimeric series, e.g. haemanthamine (49), a larger coupling 

between H-2 and H-3 ( J 2,3 5 Hz) is shown, the coupling between H-1 and H-3 

not being detectable. 

3 It is frequently possible to observe an additional coupling of H-2 with the 

equatorial H-4β, in a W-mechanism. 

4 The proton H-4α shows a large coupling with H-4a ( J 4∝,4a ~ 13 Hz) due to their 

trans-diaxial position, characteristic of the haemanthamine series. 

5 Two doublets for an AB system, corresponding to the benzylic protons of 

position C-6. 

6 The pairs of alkaloids with a hydroxy substituent at C-6, like papyramine/ 

6-epipapyramine (44/45), haemanthidine/6-epihaemanthidine (51/52), etc., 

appear as a mixture of epimers not separable even by HPLC. 

7 Also in relation with position C-6, it is interesting to note that ismine (67), a 

catabolic product from the haemanthamine series, shows a restricted rotation 

around the biarylic bond, which makes the methylenic protons at the benzylic 

position magnetically non-equivalent. 

Tazettine type 

Although tazettine (58) is one of the most widely distributed alkaloids in the 

Amaryllidaceae family, it was found to be an extraction artefact of pretazettine (60) 

(Wildman and Bailey, 1967). 

The presence of an N-methyl group (2.4–2.5 ppm) in tazettine type alkaloids 

immediately distinguishes them from the haemanthamine type, from which they 

proceed biosynthetically. Moreover the 1 H-NMR spectrum always shows the signal 

corresponding to the methylenedioxy group.


We have also included the alkaloid obesine (62) in this group, although it exhibits 

some structural differences with the skeleton type. 


Among the Amaryllidaceae alkaloids, only the galanthamine type shows an orthocoupling 

constant between both aromatic protons of ring A. The general characteristics 

of their 1H-NMR spectra are: 

1 Two doublets for the two ortho-oriented aromatic protons with a coupling 

constant of J 7,8 ~8Hz. 

2 The assignment of the substituent stereochemistry at C-3 is made in relation 

with the coupling constants of the olefinic protons H-4 and H-4a. When 

coupling constant J 3,4 is about 5 Hz, the substituent is pseudoaxial, while if it is 

~0 Hz this indicates that the substituent at C-3 is pseudoequatorial. 

3 Two doublets as an AB system corresponding to the benzylic protons of C-6. 

4 The existence of the furan ring results in a deshielding effect in H-1. 

5 This type of alkaloid often shows an N-methyl group but occasionally N-formyl 

or N-acetyl derivatives have been reported. 

Carbon 13 nuclear magnetic resonance 

13 C-NMR spectroscopy has been extensively used for determining the carbon 

framework of Amaryllidaceae alkaloids, and the major contributions are due to 

Crain et al. (1971), Zetta et al. (1973) and Frahm et al. (1985). The assignments are 

made on the basis of chemical shifts and multiplicities of the signals (by Distorsionless 

Enhancement by Polarisation Transfer (DEPT) experiment). The use of 2D-NMR 

techniques such as Heteromolecular Multiple Quantum Correlation (HMQC) and 

Heteromolecular Multiple Bond Correlation (HMBC) allows the assignments to 

be corroborated. 

The 13 C-NMR spectra of narcissus alkaloids can be divided in two regions. The 

low-field region (>90 ppm) contains signals of the carbonyl group, the olefinic and 

aromatic carbons as well as that of the methylenedioxy group. The other signals, 

corresponding to the saturated carbon resonances, are found in the high-field 

region, the N-methyl being the only characteristic group, easily recognisable by a 

quartet signal between 40–46 ppm. 

The effect of the substituent (OH, OMe, OAc) on the carbon resonances is of 

considerable importance in localising the position of the functional groups. 

The analysis of the spectra allows conclusions to be drawn about the following 

aspects: 

1 The number of methine olefinic carbons. 

2 The presence and nature of the nitrogen substituent. 

3 The existence of a lactonic carbonyl group. 

4 The presence of a quaternary carbon signal assignable to C-10b in the chemical 

shift range of 42–50 ppm.


It is worth noting that the montanine type alkaloids are distinguished from the 

other ring type systems by the downfield signal at around 150 ppm, assignable to 

the quaternary olefinic C-11a. 

Mass Spectrometry 

Extensive studies on the Mass Spectrometry of Amaryllidaceae alkaloids by electron 

impact were reported in the 1960s and 1970s (Fales et al., 1969, 1970; Ibuka et al., 

1966; Onyiriuka and Jackson, 1978; Samuel, 1975). The fragmentation patterns 

in the Electronic Impact Mass Spectrometry (EIMS) of various skeletal types are 

fairly well documented and have considerable diagnostic value. 

Lycorine type 

The molecular ion appears as a quite intense peak, and generally suffers the loss of 

water, as well as C-1 and C-2 and their substituents by a retro Diels-Alder fragmentation 

(Figure 6.13). The loss of water is not present in the spectra of acetyl 

derivatives. 

The ease of the loss of water from the molecular ion was found to be greatly 

dependent on the stereochemistry of the C-2 hydroxyl group. Thus, in the mass 

spectrum of lycorine (1), the relative intensity is low, while in 2-epilycorine it is the 

base peak (Kinstle et al., 1966). 

Homolycorine type 

In this structure type, the cleavage of the labile bonds in ring C by a retro 

Diels-Alder reaction is dominant, generating two fragments: one, the more characteristic, 

represents the pyrrolidine ring (plus substituents in position 2), and the 

other (a less abundant fragment) encompasses the aromatic lactone or hemilactone 

moiety (Figure 6.14). 

Figure 6.13 Mass fragmentation pattern of lycorine (1).

Figure 6.14 Mass fragmentation pattern of homolycorine (23). 


A further general and noteworthy feature is the low abundance of the molecular 

ion in all compounds with a double bond ∆ 3,4 (Schnoes et al., 1968). 

Crinine-haemanthamine types 

Several general considerations should be taken into account for these types of 

alkaloids: 

1 The stability of the molecular ion, which is almost always the base peak. 

2 The important role played by the aromatic ring in the stabilisation of the ions, 

which is retained in all fragments of high mass while the nitrogen atom is often 

lost. 

3 The relatively large number of nitrogen-free ions. 

4 The fragmentation mechanisms are initiated by the rupture of a bond β to the 

nitrogen atom which implies the opening of the C-11/C-12 bridge (Longevialle 

et al., 1973a,b). 

(a) Compounds with a saturated ring C and no bridge substituent. 

The configuration of the substituent on ring C plays a minor role in the fragmentation 

process. 

(b) Compounds with a double bond (∆ 1,2 ) in ring C and no bridge substituent. 

The fragmentation pattern involves ruptures of C-4a/C-10b and C-3/C-4 

bonds. A characteristic feature is the loss of a nitrogen-containing moiety, 

C3H5N [M-55]. 

(c) Compounds with a double bond (∆1,2 ) in ring C and a hydroxyl substituent at 

C-11.


Figure 6.15 Mass fragmentation pattern of haemanthamine (49). 

The presence of a hydroxyl group on C-11 is responsible for dramatic changes 

in the fragmentation pattern (Figure 6.15), and it is profoundly influenced by 

the stereochemistry. There are three fundamental patterns of fragmentation: 

1 Loss of CH 3 OH: it is more favourable when the two-carbon bridge and the 

substituent on C-3 are on the same side of the molecule. 

2 Loss of C 2 H 6 N: the relative significance of the loss of this neutral nitrogen 

moiety is governed by the ease with which the methanol is eliminated. 

3 Loss of CHO: A peak at m/z [M-29] due to the loss of an aldehyde radical is 

present in all compounds of this type. 

Tazettine type 

Minor changes in stereochemistry are sufficient to cause appreciable differences in 

the stereoisomers in these kind of structures. Thus, in the MS of tazettine (58), 

with a β configuration of the methoxyl group at C-3, the dominant ion occurs at 

m/z [M-84], following a C-ring fragmentation by a retro Diels-Alder process. 

In contrast, the mass spectrum of criwelline (59), which differs only in the 

configuration of the mentioned methoxyl group, contains a peak of low abundance 

at m/z [M-84] (Figure 6.16). Ions occur in both steroisomers due to the 

successive loss of a methyl radical and water from the molecular ion (Duffield 

et al., 1965).


Figure 6.16 Mass fragmentation pattern of tazettine (58) and criwelline (59). 

Montanine type 

The mass spectral fragmentation patterns observed for alkaloids containing the 

5,11-methanomorphanthridine nucleus is very depending of the substituents at 

C-2 and C-3, the nature as well as their particular configuration have a very significant 

effect. Thus, all the compounds which possess a methoxyl group give rise to 

an M-31 ion. 

The configuration of the C-2 substituent has a considerable effect on the extent 

to which the retro Diels-Alder fragmentation ion is observed (Figure 6.17). There 

Figure 6.17 Mass fragmentation pattern of montanine (85).


is a definite enhancement of this fragmentation when the C-2 substituent has an α 

configuration (Wildman and Brown, 1968). 


In this type of structures, the intense molecular ion, as well as [M-1] peak, the 

breaking of ring C (losing a C4H6O fragment) and the elimination of elements of 

ring B (including the nitrogen atom) are characteristic (Figure 6.18). This behaviour 

is similar for the dihydro derivatives (Razakov et al., 1969). 

Figure 6.18 Mass fragmentation pattern of galanthamine (69). 

BIOLOGICAL AND PHARMACOLOGICAL ACTIVITIES 

Toxic and hallucinogenic effects 

Plants of this genus have been used throughout history as a stimulant to induce 

trance and hallucinations, and as an agent in suicide. It has been known for a long 

time that daffodil ingestion is very dangerous, resulting in toxic symptoms in both 

man and animals (Jaspersen-Schib et al., 1996; Wilson, 1924; Wu et al., 1965). After 

ingestion of N. pseudonarcissus or N. jonquilla (Vigneau et al., 1984), the first visible 

symptoms are salivation, acute abdominal pains, nausea, vomiting and diarrhoea, 

followed by neurological (trembling, convulsions, etc.) and cardiac sequel, and 

sometimes resulting in death if eaten in larger quantities. There are many cases of 

poisoning in which daffodil bulbs were cooked by mistake in the place of leeks or 

onions. The bulbs of N. poeticus are more dangerous than those of N. pseudonarcissus, 

being powerfully emetic and irritant. In turn, N. papyraceus is believed to be toxic 

for herbivorous mammals; in this case, the alkaloid content is five times higher in 

the aerial part than in the bulbs (Suau et al., 1990a). The good news is that the 

bulb tastes awful, making it highly unlikely that anyone could keep down even one 

bite. In cases of massive ingestion, activated charcoal, salts and laxatives are adminis-


tered. When symptoms are severe, atropine sulphate is given by intravenous 

injection ( Junko et al., 1994). 

Some Narcissus species can produce harmful effects without being swallowed. 

Thus, species like N. bulbocodium must not be placed in confined spaces because 

the scent of the flowers, when present in any quantity, can produce headaches and 

even vomiting. The association of alkaloids with essential oils is found in oils of 

patchouli, juniper, orange and jonquil absolute (Maurer, 1994). In turn, N. pseudonarcissus 

shows irritant and allergenic properties on contact with animals and men 

(Bruynzeel, 1997; Bruynzeel et al., 1993; De Jong et al., 1998; Gonçalo et al., 1987; 

Güneser et al., 1996). The compounds responsible for the irritation are not known, 

but alkaloids are thought to be involved (Gude et al., 1988). Additionally, extracts 

of the bulbs, when applied to open wounds, can produce staggering, numbness of 

the whole nervous system and paralysis of the heart. The mucilage secreted by 

bulbs can also produce harmful effects in plant species such as rose, rice and cabbage, 

inhibiting seed germination and seedling growth (Bi et al., 1998; Van Doorn, 

1998). 

Traditional medicinal usage 

Despite their lethal potential, the extracts of various narcissus plants have been 

used in traditional medicine to treat a variety of medicinal problems. This aspect is 

covered in Chapter 2 of this volume. 

Biological activities for extracts of Narcissus 

Several Narcissus extracts have shown the following activities: antiviral (Abou- 

Karam and Shier, 1990; Furusawa et al., 1973, 1975; Papas et al., 1973; 

Ramanathan et al., 1968; Suzuki et al., 1974; Vacik et al., 1979; Van den Berghe et al., 

1978), prophage induction (Dornberger and Lich, 1982), antimicrobial (Dornberger 

and Lich, 1982; Ieven et al., 1979), antifungal (Chaumont and Senet, 1978; 

Chaumont et al., 1978), cytotoxic (Abou-Karam and Shier, 1990; Furusawa et al., 

1973, 1980), antitumour (Furusawa et al., 1972, 1973, 1975; Papas et al., 1973; 

Suzuki et al., 1974; Wu et al., 1965), antimitotic (Ikram, 1983), hipotensive (Chiu et al., 

1992), emetic (Wu et al., 1965), antifertility (Matsui et al., 1967), antinociceptive 

(Çakici et al., 1997), chronotropic (Chiu et al., 1992), pheromone (Keiser et al., 1975), 

plant growth inhibitor and allelopathic (Bi et al., 1998; Ceriotti, 1967; Chiu et al., 

1992; Van Doorn, 1998). 

Biological and pharmacological activities of Narcissus compounds 

The compounds responsible for the majority of the above-mentioned activities are 

the alkaloids. However, the mannose-binding lectins have also received much 

interest recently (Barre et al., 1996; Van Damme et al., 1998). 

The different pharmacological and/or biological properties exhibited by the 

alkaloids from the genus Narcissus are shown in Table 6.4, but only some of the 

activities of a reduced number of these alkaloids are known. The most extensively 

studied effect is that of non-specific inhibition. The relationship of chemical structure


and biological activity is largely unknown (Chiu et al., 1992), and further studies of 

several alkaloids still remain to be done for therapeutic purposes. The best-studied 

alkaloids in this group are galanthamine, lycorine, narciclasine and pretazettine, 

which possess a diversity of pharmacological activities. 

Table 6.4 Biological and pharmacological activities of Narcissus alkaloids 

Alkaloid Activity References 

10 caranine * Weak analgesic 

* Convulsant and hypotensive 

* Acetylcholinesterase inhibitor 

* Active against the murine 

P-388 lymphocytic leukaemia (as 

acetylcaranine) 

53 crinamine * Powerful transient hypotensive 

in dogs 

* Respiratory depressant 

* Moderate antimalarial 

* Antimicrobial 

* Moderately active against 

murine Rauscher viral 

leukaemia 

* Cytotoxic against Molt 4 

lymphoid and LMTK 

fibroblastic cell lines 

* Weak cytotoxic against HepG2 

hepatoma 

36 dubiusine * Cytotoxic against non-tumoural 

LMTK cells 

* Moderately active against Molt 4 

lymphoma 

69 galanthamine * Powerful analgesic as strong as 

morphine 

* Anticonvulsive 

* Hypotensive 

* Inductor of hypothermia in rat 

* Acetylcholinesterase inhibitor 

with peripheral and central 

pharmacological effects 

* Highly selective for 

acetylcholinesterase versus 

butyrylcholinesterase (more than 

50-fold greater) 

* Centrally-acting 

acetylcholinesterase inhibitor 

which has shown potential for 

the treatment of Alzheimer’s 

disease 

* Attenuates or reverses cognitive 

deficits induced by drugs – and 

lesions – in animal models 

* Acts as noncompetitive nicotinic 

receptor agonist 

CHCD, 1996 

Pettit et al., 1984 

CHCD, 1996 

Furusawa et al., 1980 

Likhitwitayawuid et al., 1993 

Weniger et al., 1995 


Bores et al., 1996 

Chao et al., 1965 

CHCD, 1996 

Cozanitis, 1977 

Cozanitis and Rosenberg, 1974 

Cozanitis and Toivakka, 1978 

Cozanitis et al., 1983 

Fulton and Benfield, 1996 

Ghosal et al., 1985b 

Han et al., 1992 

Harvey, 1995 

Kewitz, 1996 

Mihailova and Yamboliev, 

1986 

Mihailova et al., 1985 



Pereira et al., 1994 

Radicheva et al., 1996 

Riemann et al., 1994 

Schuh, 1976 

Storch et al., 1995 

Suzuki et al., 1974



* Combines both physostigmine 

and neostigmine properties: 

* Like physostigmine, reverses 

opioid-induced respiratory 

depression, but not the 

concomitant analgesia 

* Like neostigmine, 

antagonizes muscle paralysis 

induced by d-tubocurarine, 

also antagonizes the ganglionic 

blockade and increases 

the contraction response 

* As hydrobromide has shown 

several central and peripheral 

effects: 

* Has central effects such as 

antagonism of the respiratory 

depressant effect of 

morphine-like compounds 

* Is capable of penetrating the 

blood-brain barrier and it is 

used in the treatment of the 

central effects of scopolamine 

(hyoscine) intoxication. 

It has certain advantages over 

physotigmine for this purpose 

* Shortens REM latency, 

increases REM density, and 

reduces slow wave sleep 

* Is able to reverse the central 

anticholinergic syndrome 

* used for its anticholinesterase 

activity in the treatment of 

disturbances in the peripheral 

sympathetic synaptic 

transmission 

* Reverses the neuromuscular 

blocking effect of curare-type 

muscle relaxants and has been 

used safely in anaesthesia 

(postsurgery) and in the 

treatment of various 

neurologic disorders (pareses, 

paralysis of different origins, 

myasthenia gravis, progressive 

muscular dystrophy, etc.) 

* Affects the contraction of 

skeletal muscle not only at the 

level of neuromuscular 

junction, but also by enhancing 

the efferent impulses as a result 

of the reflexive excitation 

centre in the spinal cord 

Svensson and Nordberg, 1997 

Sweeney et al., 1988 

Sweeney et al., 1989 

Thomsen and Kewitz, 1990 



Westra et al., 1986 

Zakirov and Umarova, 1971




(contd.) 

* Is over 90% bioavailable after 

oral administration and has a 

plasma elimination half-life of 

approximately 6 hours. 

* Nausea and vomiting are the 

most commonly reported 

adverse effects; liver toxicity has 

not been reported to date 

* Energix®, a preparation 

composed of galanthamine, 

increases endurance during 

exercise and delays the onset 

of fatigue 

* Used in a pharmaceutical 

composition for treatment of 

alcoholism 

* Cytotoxic against non-tumoural 

LMTK cells 

* Produces poisoning of digestive, 

respiratory, neuromuscular 

and central nervous systems 

70 epigalanthamine * More hypotensive and less 

toxic than galanthamine 

* Anticholinesterase activity 

lower than galanthamine 



Thomsen et al., 1990 

Yamboliev et al., 1988 

Zakirov and Umarova, 1971 


72 norgalanthamine * Cytotoxic against Molt 4 lymphoid 

and LMTK fibroblastic cell lines 

74 N-formyl- * Moderate cytotoxic against Weniger et al., 1995 

norgalanthamine Molt 4 lymphoma and HepG2 

hepatoma 

* Cytotoxic against non-tumoural 

LMTK cells 

7 galanthine * Analgesic 

CHCD, 1996 

* Hypotensive 


* Weak cytotoxic against Molt 4 

lymphoid cells 


49 haemanthamine * Hypertensive 

Báez and Vázquez, 1978 

* Cytotoxic against a variety of Codina et al., 1992b 

cultured cells (in vitro) 


* Cytotoxic against fibroblastic Ghosal et al., 1985b 

LMTK cells 

Jiménez et al., 1975b 

* Moderately active against Molt 4 Jiménez et al., 1976 


Lin et al., 1995 

* Moderately active against 

Rauscher viral leukaemia 

* Inhibitor of HeLa cell growth 

* Inhibitor of protein synthesis, 

blocking the peptide bond 

formation step on the peptidyl 

transferase centre of the 60S 

ribosomal subunit 

Weniger et al., 1995



* Slightly reduces DNA synthesis, 

whereas RNA synthesis is 

practically unaffected 

51 haemanthidine * Analgesic 

* Antiinflamatory 

* Active against A-431, KB, Lu1, 

Me12 and ZR-75-1 cell lines 

* Significantly active against 

LNCaP and HT cell lines 

30 hippeastrine * Antiviral. Active against Herpes 

simplex type 1 

* Weak insect antifeedant 

* Significantly active against the 

LNCaP and HT cell lines 

* Cytotoxic against non-cancerous 

LMTK cells 



23 homolycorine * Inductor of delayed 

hypersensitivity in animals 

* Cytotoxic against fibroblastic 

LMTK cells 


29 9-O-demethyl- 

2α- hydroxyhomolycorine 


LMTK cells 




LMTK cells 

* Moderately active against Molt 4 


67 ismine * Cytotoxic against Molt 4 

lymphoid and LMTK 

fibroblastic cell lines 

77 lycoramine * Central anticholinesterase activity 

stronger than galanthamine 

* Inhibits the formation of peptide 

bond in protein synthesis 

* Like neostigmine, antagonizes 

muscle paralysis induced by 

d-tubocurarine, also antagonizes 

the ganglionic blockade, and 

increases the contraction 

response 

* Produces acute poisoning 

of digestive, respiratory, 

cardiovascular, neuromuscular 

and central nervous systems 

31 lycorenine * Analgesic 

* Weak hypotensive 

* Vasodepressor action ascribed to 


Çitoglu et al., 1998 

Tanker et al., 1996 


Renard-Nozaki et al., 1989 







Chao et al., 1965 

Missoum et al., 1997 


Codina et al., 1992b 


Miyasaka and Hiramatsu, 

1980



31 lycorenine 

(contd.) 

maintenance of its α-adrenergic 

blocking action in conjugation 

with the reduction of the spontaneous 

sympathetic nerve activity, 

and produce bradycardia by 

Miyasaka et al., 1979 


32 O-methyl- 

modifying vagal activity 

* Cytotoxic against murine LMTK 

and human HepG2 cell lines 

* Cytotoxic against fibroblastic Weniger et al., 1995 

lycorenine 

LMTK cells 


hepatoma 

1 lycorine * Emetic 

Abbassy et al., 1998 

* Analgesic 

Abdalla et al., 1993 

* Antiinflammatory 

Arrigoni et al., 1975 

* Respiratory stimulant 

Arrigoni et al., 1996 

* Expectorant 

Arrigoni et al., 1997a 

* Used to treat bronchitis and Arrigoni et al., 1997b 

bronchial asthma 


* Relaxant of isolated 

Campbell et al., 1998 

epinephrine-precontracted Carrasco et al., 1975 

pulmonary artery 

Chattopadhyay et al., 1984 

* Increases contractility and rate CHCD, 1996 

of isolated perfused heart. These Çitoglu et al., 1998 

effects are mediated by stimulation Córdoba-Pedregosa et al., 1996 

of β-adrenergic receptors Davey et al., 1998 

* Specific inhibitor of ascorbic acid De Gara et al., 1994 

biosynthesis. Seems to act as a Del Giudice et al., 1997 

powerful inhibitor of the mitocon- Evidente et al., 1983a 

drial L-galactono-γ-lactone Evidente et al., 1983b 

dehydrogenase 


* Most of the effects of lycorine on Evidente et al., 1986 

physiological processes have Gabrielsen et al., 1992 

been ascribed to its ability to Ghosal et al., 1984 

inhibit ascorbic acid 


biosynthesis in vivo 

Ghosal et al., 1985c 

* The ability to inhibit the ascorbic Ghosal et al., 1988b 

acid biosynthesis has made this Ghosal et al., 1989b 

substance a valuable tool for Ghosal et al., 1990 

studying the ascorbic acid Ieven et al., 1982 

dependent reactions 

Ieven et al., 1983 

* Inhibitor of cyanide-resistant Jiménez et al., 1975b 

respiration. Ascorbic acid is Jiménez et al., 1976 

needed for the synthesis of Kushida et al., 1997 

hydroxyproline-containing Likhitwitayawuid et al., 1993 

proteins, specifically utilised Lin et al., 1995 

for the development of 

Liso et al., 1984 

KCN-resistant respiration Liso et al., 1985 

* Inhibitor of peroxidase 

Makhkamova and Safonova, 1994 

enhancement, which seems Massardo et al., 1994 

related with the synthesis of Papas et al., 1973 

hydroxyproline-containing Renard-Nozaki et al., 1989 

proteins 

Schultz et al., 1996



* Powerful inhibitor of growth and 

cell division in higher plants, 

algae and yeasts, inhibiting the 

cell cycle during interphase. 

Ascorbic acid is required for cell 

division 

* Inhibitor of cell division in rat 

fibroblasts. Ascorbic acid is 

required for cell division 

* Inductor of flat morphology in 

K-ras-NRK cells (transformed 

fibroblasts) 

* Inhibitor of protein synthesis, 


formation step 

* Plant growth inhibitor by 

inhibition of protein synthesis 

* The effects on ascorbic acid 

biosynthesis occur at 

concentrations below those at 

which protein synthesis is 

affected 

* Moderate antitumoural. Its 

mechanism of action is thought 

to be through inhibition of 

protein synthesis at the 

ribosomal level 

* Decreases the growth of several 

viruses through its inhibitory 

action on viral protein synthesis 

* Active against several RNA 

and DNA viruses 

* Does not inhibit the activity of 

reverse transcriptase 

* Inhibitor of DNA synthesis 

* Is able to differentiate between 

cells containing mitocondrial 

DNA and those lacking it. 

Inhibits growth of strains 

containing mitocondrial DNA 

* Cytotoxic against a variety of 

cultured cell lines 

* Inhibitor of HeLa cells growth 

* Antimalarial 

* Weak protozoicide 

* Inhibitor of germination 

of seeds and growth of roots. 

Lycorine-1-O-β-D-glucose has 

the reverse effect, and may 

also produce mitogenic activity 

in animal cells 

* Ungeremine, a natural 

metabolite of lycorine, is 

Singh and Pant, 1980 

Tanker et al., 1996 

Van den Berghe et al., 1986 

Vrijsen et al., 1986 


Yui et al., 1998



1 lycorine 

(contd.) 

responsible, at least in part, 

for the growth-inhibitory and 

cytotoxic effects of lycorine 

* Certain bacterias transform 

lycorine into pancrassidine 

(less cytotoxic than ungeremine) 

* The changes observed in 

response to stress suggest 

its role in protective and repair 

mechanisms of producer plants 

* Insecticide 

* Insect antifeedant 

61 3-epimacronine * Weak cytotoxic against human 

Molt 4 and murine LMTK cell 

lines 


40 maritidine * Antineoplastic Alarcón et al., 1986 


27 masonine * Inductor of delayed 

hypersensitivity in animals 


82 mesembrenone * Cytotoxic against Molt 4 


* Weak cytotoxic against 

fibroblastic LMTK cells 


63 narciclasine * Antimitotic and cell growth 

inhibitor 

* Strong tumour inhibitor. 

One of the most important 

antineoplastic Amaryllidaceae 

alkaloids 

* Active against larynx and 

cervix carcinomas 

* Inhibitor of growth of Ehrlich 

tumour cells 

* Inhibitor of HeLa cells growth 

* No effect has been observed 

towards solid tumours 

(sarcoma 180) 

* Inhibitor of protein synthesis 

in eukariotic ribosomes, 


formation on the 60S ribosomal 

subunit. 

* Resistance to narciclasine in a 

mutant strain of Saccharomyces 

cerevisiae is due to an alteration 

on the peptidyl transferase 

centre 

* Reduces DNA synthesis, whereas 

RNA synthesis is practically 

unaffected 

* Enhances the uptake of uridine 

and stimulates the synthesis of 

Abou-Donia et al., 1991 

Aller, 1981 


Bi et al., 1998 

Carrasco et al., 1975 


CHCD, 1996 



Gabrielsen et al., 1992 

Ghosal et al., 1989a 

Hua et al., 1997 

Jiménez et al., 1975a 


Jiménez et al., 1976 

Keck and Fleming, 1978 



Pettit et al., 1995a 

Pettit et al., 1995b 

Rodríguez-Fonseca et al., 1995 

Veronese et al., 1991



pre-rRNA (38S) and 

low-molecular weight 

(4–5 S) RNA 

* Inhibitor of ascorbic acid 

biosynthesis in vivo 

* Inhibitor of cell division in 

plant tissue cultures 

* Inhibitor of seed germination 

and seedling growth in a 

dose-dependant manner, 

interacting with hormones in 

some physiological responses. 

The O-glucoside of narciclasine 

has the reverse effect 

* Strong antibiotic. Active against 

Corynebacterium fascians 

* Active against RNA-containing 

flaviviruses and bunyaviruses 

* Narciclasine-4-O-β-Dglucopyranoside 

has shown 

cytotoxic and antitumour 

activity very similar to narciclasine 

* The peculiar activity of 

narciclasine seems to arise from 

the functional groups and 

conformational freedom of 

its C-ring 

76 narwedine * Hypotensive 

* Increases the amplitude and 

frequency of respiratory 

movements 

* Increases the amplitude and 

decreases the frequency of 

cardiac contractions 

* Decreases soporific effects of 

ethanol and barbiturics 

* Increases the analgesic effect 

of morphine 

* Protective against thiopental 

poisoning 

* Enhances the effects of caffeine, 

corazole, arecoline, and nicotine 

* Evaluated as potential agent for 

treatment of Alzheimer’s disease 

* Probably acts primarily on 

m-cholinoreactive structures 

of the brain 

44 papyramine * Cytotoxic against fibroblastic 

LMTK cells 

* Weak cytotoxic against human 

tumoural cell lines Molt 4 and 

HepG2 



Szewczyk et al., 1995 

Zakirov and Umarova, 1971 

Weniger et al., 1995



60 pretazettine * Analgesic 

* Anticancer activity 

* Active against murine Rauscher 

viral leukaemia 

* Active against spontaneous AKR 

T cells in mice 

* Active against intraperitoneally 

and subcutaneously implanted 

LLC (Lewis lung carcinomas) 

* Effective against Ehrlich ascites 

tumour, a non viral 

transplantable tumour 


LMTK and Molt 4 lymphoid 

cell lines 


* Inhibitor of protein synthesis in 

eukariotic ribosomes, blocking 

the peptide bond formation on 

the 60S ribosomal subunit. 

* Slightly reduces DNA synthesis; 

has no effect on RNA synthesis 

* Active against Herpes simplex 

type 1 (HS-1) 

* Active against neurotropic RNA 

viruses 

* Potent inhibitor of viral reverse 

transcriptase of RNA tumour 

viruses, binding to the 

polymerase enzyme 

* Demonstrates synergistic effect 

in combination with standard 

cytotoxic drugs 

* In combination with ryllistine 

inhibits nucleic acid synthesis 

* Can be administrated over a long 

period of time without apparent 

toxicity 

3 pseudolycorine * Effective against murine 

Rauscher leukaemia in mice 

without apparent toxicity 

* Inhibitor of protein synthesis 

in tumour cells at the step of 

peptide bond formation. It has 

a different binding site than 

lycorine on the peptidyl 

transferase centre of the 60S 

ribosomal subunit 

* Reduces DNA synthesis. RNA 

synthesis is practically unaffected 

* Cytotoxic against human Molt 4 

and murine LMTK cells 



CHCD, 1996 

Furusawa and Furusawa, 1986 

Furusawa and Furusawa, 1988 


Furusawa et al., 1976b 







Ghosal and Razdan, 1984 






Rigby et al., 1998 

Suzuki et al., 1974 

Van den Berghe et al., 1986 


Zee-Cheng et al., 1978 









Papas et al., 1973 


Suzuki et al., 1974 


Zee-Cheng et al., 1978




* Moderately active against HepG2 

hepatoma 

* Moderately active against Ehrlich 

ascites tumour cells 


* Antiviral. Active against Herpes 

simplex type 1 

* Active against neurotropic RNA 

viruses 

* Does not inhibit the activity of 

reverse transcriptase 

* Cytotoxic against Molt 4 lymphoid 

and LMTK fibroblastic cell lines 


hepatoma 

58 tazettine * Weak hypotensive 

* Exhibits mild activity against 

certain tumour cell lines 

* Active against the Co12 cell line 

* Weak cytotoxic against 

fibroblastic LMTK cell lines 

* The stereochemical 

rearrangement from pretazettine 

to tazettine inactivates the 

biological activity of pretazettine 

39 vittatine * Weak analgesic in mice 

* Increases the analgesic effect of 

morphine 

* Tachycardic in dogs 




CHCD, 1996 

Codina et al., 1992b 

Furusawa et al., 1976b 


Rigby et al., 1998 


CHCD, 1996 


Galanthamine, originally isolated from Galanthus nivalis in the 1940s, is a longacting, 

selective, reversible and competitive inhibitor of acetylcholinesterase. This 

enzyme is responsible for the degradation of acetylcholine at the neuromuscular 

junction, in peripheral and central cholinergic synapses and in parasympathetic 

target organs (Fulton and Benfield, 1996; Wilcock and Wilkinson, 1997). Galanthamine 

hydrobromide was first used by Bulgarian and Russian researchers in the 

1950s and has been exploited for a variety of clinical purposes in the past. It has been 

used clinically for postsurgery reversal of tubocurarine-induced muscle relaxation 

and for treating post-polio paralysis, myasthenia gravis and other neuromuscular 

diseases, as well as traumatic brain injuries (Bores and Kosley, 1996; Radicheva 

et al., 1996). Besides this, galanthamine acts as a mild analeptic, shows analgesic 

power as strong as morphine, and, applied in eye drops, reduces the intraocular 

pressure. As early as 1972, Soviet research had demonstrated that galanthamine 

could reverse scopolamine-induced amnesia in mice, a finding that was demonstrated 

in man four years later. However, this did not lead to the application of 

this compound in Alzheimer’s disease until 1986, long after the widely accepted 

cholinergic hypothesis of Alzheimer’s disease had been first postulated (Allain 

et al., 1997). While the original acetylcholinesterase inhibitors, physostigmine and


tacrine, left much to be desired, galanthamine hydrobromide offers superior 

pharmacological profiles and increased tolerance (Ezio, 1998; Herman and 

Mucke, 1997a; Kewitz, 1997; Nordberg and Svensson, 1998; Rainer, 1997a). 

From the clinician’s point of view, galanthamine is a reasonable approximation of 

the ideal concept of symptomatic Alzheimer’s disease therapy (Rainer, 1997b; 

Wilcock and Wilkinson, 1997). Until very recently galanthamine could only be 

obtained in very small amounts from plant material (mainly from Leucojum aestivum) 

as its total chemical synthesis on an industrial scale was uneconomical (Eichhorn 

et al., 1998; Hermann and Mucke, 1997b). This strictly limited availability of 

galanthamine was a severe constraint and probably the main reason for the 

cautious approach taken by the international pharmaceutical community. It appears 

that this situation has now fundamentally changed, as reflected in the increasing 

number of scientific reviews concerned exclusively with galanthamine and its 

derivatives (Bores and Kosley, 1996; Bores et al., 1996; Brodaty, 1996; Fulton and 

Benfield, 1996; Harvey, 1995; Kewitz, 1996, 1997; Mucke, 1997a,b; Nordberg and 

Svensson, 1998; Svensson and Nordberg, 1997). This has resulted in much interest 

in the chemical synthesis of the product on an industrial scale (Czollner et al., 1998; 

Kita et al., 1998; Szewczyk et al., 1995), and in production by in vitro tissue cultures, 

mainly from Narcissus confusus (Bergoñón et al., 1996; Sellés et al., 1997, 1999). 

Lycorine, the most frequent and characteristic of Amaryllidaceae alkaloids, has 

been reported to be a powerful inhibitor of L-asc (ascorbic acid) biosynthesis 

(Arrigoni et al., 1975; Evidente et al., 1983b), and thus has been proved to be a 

useful tool in studying asc-dependent metabolic reactions in asc-synthesising 

organisms (Arrigoni et al., 1997a). Lycorine is actually a powerful inhibitor of the 

activity of GL dehydrogenase (L-galactono-γ-lactone dehydrogenase), the terminal 

enzyme of asc biosynthesis (Davey et al., 1998; De Gara et al., 1994), which appears 

to be localised in the mitochondrial membrane (Arrigoni et al., 1996, 1997b). 

Administration of lycorine induces a decrease in asc content and a simultaneous 

increase of dehydroascorbic acid levels in plants (De Tullio et al., 1998). It has been 

suggested that the alkaloid could act as an inhibitor of asc biosynthesis in vitro, 

without interfering with asc utilisation in cells (Arrigoni et al., 1997b). The effects 

of lycorine on L-asc biosynthesis have been reported to occur at concentrations 

below those at which protein synthesis is affected, but it seems difficult to rule out 

completely non-specific effects of this alkaloid since it has been reported that, at 

least in yeasts, lycorine is able to interact directly with mitocondrial DNA (Davey 

et al., 1998; Del Giudice et al., 1997; Massardo et al., 1994). It is also well documented 

that lycorine is a powerful inhibitor of cell growth, cell division and organogenesis 

in higher plants, algae and yeasts, which seems to be related with asc levels 

(Arrigoni, 1994; Arrigoni et al., 1997a; Córdoba-Pedregosa et al., 1996; Del 

Giudice et al., 1997; Onofri et al., 1997). Lycorine also shows antitumour activity 

(Yui et al., 1998). Its mechanism of action is thought to be through inhibition of 

protein synthesis at the ribosomal level (Furusawa et al., 1980; Hua et al., 1997). 

Additionally, it has antiviral (Gabrielsen et al., 1992), antifeedant (Singh and Pant, 

1980), and antimalarial as well as anti-inflammatory (Campbell et al., 1998; Çitoglu 

et al., 1998) activities. There has also been much recent interest in the chemical 

synthesis of lycorine (Hoshino et al., 1996; Ishizaki et al., 1998; Schultz et al., 1996). 

Narciclasine, an antimitotic and antitumoural alkaloid from Narcissus bulbs (Ceriotti, 

1967), inhibits protein synthesis by directly interacting with the 60S ribosomal


subunit and inhibiting peptide bond formation by preventing binding of the 3′ 

terminal end of the donor substrate to the peptidyl transferase centre (Carrasco et al., 

1975). Narciclasine also inhibits seed germination and seedling growth of some 

plants in a dose-dependent manner, interacting with hormones in some physiological 

responses. In this way, indole-3yl-acetic acid cannot overcome the inhibition 

of elongation of wheat coleoptile sections caused by narciclasine. Additionally, 

narciclasine suppresses the gibberellin-induced α-amylase production in barley 

seeds and cytokinin-induced expansion and greening of excised radish cotyledons 

(Bi et al., 1998). There are also interesting studies related to the chemical synthesis 

of this alkaloid (Angle and Wada, 1997; Rigby and Mateo, 1997). 

The anticancer and antiviral activities exhibited by pretazettine have also stimulated 

considerable interest in this compound (Furusawa and Furusawa, 1988; Furusawa 

et al., 1980; Gabrielsen et al., 1992; Suzuki et al., 1974) and in its chemical synthesis 

(Nishimata and Mori, 1998). 

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7 Production of galanthamine 

by Narcissus tissues in vitro 

Carles Codina 

INTRODUCTION 

Galanthamine is a morphine-like alkaloid that is a possible therapeutic agent in 

Alzheimer’s disease because of its central cholinergic effects (Harvey, 1995). It has 

been shown to be competitive with other anticholinesterase compounds like 

tacrine or physostigmine in the treatment of the syndrome (Rainer et al., 1989). 

In contrast with the proven hepatotoxicity of tacrine (Gauthier et al., 1990), galanthamine 

shows only minor side effects like agitation or insommia (Thomsen et al., 

1990). Thus, galanthamine is considered a better therapeutic candidate for the 

treatment of this type of senile dementia than other acetylcholinesterase inhibitors 

structurally related to it (Bores et al., 1996). 

Galanthamine can be extracted from plants of the Amaryllidaceae family, the 

main natural sources being species of the genera Galanthus and Leucojum from 

Bulgaria and the Caucasus region. Galanthamine is difficult to obtain and extremely 

expensive for clinical usage. Although the total synthesis of galanthamine has been 

achieved (Czollner et al., 1998), the stereoselectivity of its reactions and the low 

yields make this process economically unattractive. Therefore, as the plant remains 

the only valid source of galanthamine, a biotechnological approach has been considered 

as an alternative method for the production of the alkaloid. 

Although much work has been done on the propagation of bulbous plants, very 

little exists on the production of secondary metabolites from them, this being 

restricted mainly to the production of colchicine by callus culture of Colchicum 

autumnale (Hayashi et al., 1988; Yoshida et al., 1988), haemanthamine by root cultures 

of Zephyranthes robusta (Furmanova and Oledzka, 1990), and bufadienolides 

by different tissues of Urginea indica (Jha et al., 1991). 

Investigations at the University of Barcelona with plants of the genus Narcissus 

have focused on the propagation of several species by in vitro liquid culture 

(Bergoñón et al., 1992; Bergoñón, 1994; Riera, 1996; Sellés, 1996), and also on the 

production of alkaloids using different explants of Narcissus confusus, a wild species 

which was found to contain 0.1% of galanthamine on a fresh weight basis, as well as 

other alkaloids including N-formylnorgalanthamine, haemanthamine and tazettine 

(Bastida et al., 1987). Experiments dealing with the production of galanthamine 

and related alkaloids have been performed using shoot-clumps (meristematic 

clusters), which were obtained from two different types of explants, of seed or bulb 

origin. These shoot-clumps constitute a good experimental system as they are 

made up of differentiated tissue, with a higher expression of secondary metabolism

216 C. Codina 

and, consequently, higher production of alkaloids than cell suspension cultures. 

Media were based on Murashige and Skoog (MS) medium (Murashige and Skoog, 

1962). Cultures were maintained at 25 ± 1 °C under a photoperiod of 16 h, and 

they were shaken at 110 rpm. Extraction of the alkaloids was carried out as 

described previously (Sellés et al., 1997b), and they were determined in both tissue 

and liquid medium by high performance liquid chromatography (Sellés et al., 

1997a). Results are presented of studies into the effect of cellular and tissue differentiation, 

growth regulators, sucrose concentration and culture vessel size on 

growth and production of alkaloids in Narcissus confusus explants. 

EFFECT OF THE DEGREE OF CELLULAR AND TISSUE 

DIFFERENTIATION ON ALKALOID PRODUCTION 

The results with organogenic plant material obtained by micropropagation, starting 

from both seeds and bulbs of Narcissus confusus, are presented. Although several 

experiments dealing with bud formation induced from bulb scales and from the 

basal part of the leaf of different species of Narcissus cultured in vitro have been 

published (Seabrook, 1990; Chow et al., 1993), there is little information on the 

micropropagation of these plants via callogenesis, owing to the difficulty in obtaining 

callus. The plant material used to start the experiments were seeds and bulb 

scales of dormant N. confusus. The three different types of tissues obtained from 

seeds were called friable callus, meristematic callus, and organogenic tissue, 

according to the development of the explants. In the case of bulb scales, alkaloid 

levels were determined in the shoots obtained throughout the differentiation 

process, as well as in the initial scales, separated into inner, central and outer scales 

according to their position in the bulb. 

Seed-derived plant material 

It seems that the explants offering the best potential for starting callus formation 

come from cut ovaries kept in a medium supplemented with 20 mg/l naphthaleneacetic 

acid (NAA) (Seabrook, 1990). In the present case, however, callus tissue was 

obtained from seeds. The levels of alkaloids present in the different kinds of callus 

are described below. 

Callus induction: alkaloids in friable callus 

Callus induction was carried out as previously described (Sellés et al., 1999), and 

the levels of alkaloids were determined in the two cellular strains showing the best 

growth, A, callus cultured in MS medium supplemented with 10 mg/l 2,4-dichlorophenoxyacetic 

acid (2,4-D), and B, callus maintained in MS medium supplemented 

with 10 mg/l picloram. During culture, some of the friable callus obtained 

was analysed for its alkaloid content, and the results are shown in Figure 7.1A. In 

both strains, the alkaloid profile was very similar. In general, the content of galanthamine 

was not very high at this low level of cellular differentiation, representing 

15.3–24.0% of total alkaloids. N-formylnorgalanthamine was the major alkaloid 

present, reaching values of up to 80 and 110 µg/g dry weight (DW).

Production of galanthamine in vitro 217 

Although de-differentiated callus was not the best system for alkaloid production, 

it was interesting to observe that it had a biosynthetic capability, thus constituting a 

useful experimental system for studying the biosynthesis of alkaloids because of its 

simple structure. 

Callus maintenance: alkaloids in meristematic tissue 

After 6 months of maintaining callus in the same conditions, meristematic callus 

from both strains A and B was transferred to an MS medium without auxins to 

promote embryo maturation. Two months later, the callus derived from strain A 

showed a high number of white globular structures which seemed to be cauline 

tips, and even some somatic embryo-like protuberances were observed. The B 

strain derived callus was more compact, and some of the protuberances were 

elongated like the root meristem. 

The alkaloid content was determined separately in callus from both A and B strains 

when removing the auxins from the medium. As expected, from the chemical 

point of view, the alkaloid profile was different in the two strains (Figure 7.1B), 

according to their different morphological development. While the embryogenic 

callus (strain A) accumulated mainly galanthamine-type alkaloids, N-formylnorgalanthamine 

being the most abundant (58% of total alkaloids), those from strain B 

(rhizogenic callus) contained haemanthamine as the major alkaloid (43.5% of total 

alkaloids). Thus, the type of cellular organisation influences the qualitative profile 

of alkaloids accumulated in the meristematic callus. 

Callus regeneration: alkaloids in organogenic tissue 

Callus from N. confusus has been found to maintain its regenerative capability for a 

period of about two years if kept in a medium supplemented with 10 mg/l of 2,4-D 

or picloram (Sellés, 1996). This organogenic capacity of the N. confusus callus is not 

µg/g DW 

120 

100 

80 

60 

40 

20 

0 

NFNGAL GAL HAEM TAZ 

Alkaloid 

A strain B strain 

A 

µg/g DW 

400 

350 

300 

250 

200 

150 

100 

50 

0 


Alkaloid 


Figure 7.1 Alkaloid content in friable (A) and meristematic (B) callus from two strains 

of seed-derived explants (values represent means of three replicates). NFN- 

GAL, N-formylnorgalanthamine; GAL, galanthamine; HAEM, haemanthamine; 

TAZ, tazettine. 

B

218 C. Codina 

easily lost on transfer to a regeneration medium, as happens, for instance, with 

Allium cepa (Havel and Novák, 1988). 

Regeneration of the meristematic callus (strains A and B) was induced by growing 

it in several MS culture media supplemented with 3% of sucrose and with and 

without 0.5 or 1 mg/l of the cytokinins benzyladenine (BA) and kinetin. Under 

these conditions, the globular callus became green and developed aggregates of 

young shoots, whereas numerous roots were formed in the most friable callus, 

especially in presence of cytokinins. The alkaloid content in the organogenic 

tissues was then determined in two types of aggregates, shoot and root clusters. 

Alkaloids in shoot clusters 

These organogenic tissues were constituted by aggregates of eight to ten young 

shoots per explant, although they also showed meristematic buds which were far 

less abundant in the callus grown with kinetin. The alkaloid content of these clusters 

was similar, in global terms, to that of the meristematic callus, but the alkaloid 

profile was not the same (Figure 7.2). One can observe that, in that callus with a 

higher degree of differentiation, the main alkaloid was galanthamine. 

Alkaloids in root clusters 

After seeing that the clusters derived from the B strain treated with 0.5 mg/l of 

kinetin grew well in solid medium, they were transferred to a liquid medium without 

growth regulators and supplemented with 6% of sucrose. They were maintained 

for 6 weeks, and alkaloid levels were determined every two weeks (Figure 7.3A). 

The alkaloids were progressively released into the liquid medium throughout the 

experiment. In general, the removal of alkaloids was higher in the first subculture, 

with the exception of the alkaloids haemanthamine and galanthamine, which 

reached a maximum percentage in the second and third subcultures, respectively. 

N-formylnorgalanthamine was the alkaloid released to the medium in the highest 

proportion, which was also observed in other assays carried out in liquid medium. 

µg/g DW 

200 

160 

120 

80 

40 

0 


Alkaloid 


A 

µg/g DW 

200 

160 

120 

80 

40 

0 


Alkaloid 


Figure 7.2 Alkaloid content in shoot clusters grown in MS solid medium supplemented 

with 1 mg/l of BA (A) or kinetin (B) (values represent means of three replicates). 

For abbreviations, see Figure 7.1. 

B

µ g/ml 

6.0 

5.0 

4.0 

3.0 

2.0 

1.0 

0.0 

2 4 

Time (weeks) 

6 


A 

µ g/g DW 


At the end of the experiment, the major alkaloids in both tissues and liquid 

medium were haemanthamine and N-formylnorgalanthamine, with a maximum 

content of 1.38 and 0.75 mg/g DW, respectively (Figure 7.3B). The haemanthamine 

type alkaloids were mainly accumulated in the tissue, whereas those of the galanthamine 

type were mainly released to the liquid medium, especially in the last 

subculture, coinciding with a remarkable necrosis of the root tissue. 

In general, the kind of explant used influenced the type of alkaloid obtained 

in vitro, so most of the subsequent experiments were carried out with shootclumps, 

or with small bulblets, as galanthamine has been the main alkaloid 

accumulated in these explants. 

In vitro cultured plant cells grown in an undifferentiated state either produce 

insignificant amounts of secondary metabolites or they lose their ability to produce 

them when maintained long-term. This is the case, for instance, in Urginea indica 

(Jha et al., 1991). This loss, however, can be restored when shoots or roots are 

differentiated from disorganised tissue (Payne et al., 1991). The present results 

show that embryogenic callus of N. confusus may accumulate nearly the same 

amounts of alkaloids as shoot-clumps (Table 7.1). Since the globular structures 

(embryogenic callus) which start alkaloid accumulation are rather simply organised, 

1200 

1000 

800 

600 

400 

200 

0 


Alkaloid 

tissue medium 

Figure 7.3 A: levels of alkaloids released by root clusters to the culture medium throughout 

the experiment. B: accumulation of alkaloids in both tissue (root cluster) and 

liquid medium at the end of the experiment (values represent means of three 

replicates; thin bars represent SD). For abbreviations, see Figure 7.1. 

Table 7.1 Alkaloid content (µg/g DW ± SD calculated from three replicates) at various development 

stages of Narcissus confusus tissue cultures a 

Tissue culture NFNGAL GAL HAEM TAZ 

De-differentiated callus 0.08 ± 0.02a 0.03 ± 0.02a 0.06 ± 0.00a 0.02 ± 0.00a 

Embryogenic callus 0.25 ± 0.17a 0.11 ± 0.05a 0.13 ± 0.08a 0.04 ± 0.00a 

Shoot-clumps 0.10 ± 0.01a 0.14 ± 0.01a 0.08 ± 0.00a 0.03 ± 0.00a 

Plantlets 0.73 ± 0.22b 1.43 ± 0.51b 1.02 ± 0.63b 0.68 ± 0.15b 

Note 

a means followed by the same letter are not significantly different from each other at p ≤ 0.05. Abbreviations: 

DW, dry weight; FW, fresh weight; SD, standard deviation; NFNGAL, N-formylnorgalanthamine; 

GAL, galanthamine; HAEM, haemanthamine; TAZ, tazettine. 

B

220 C. Codina 

they might be more suitable for investigating the mechanisms triggering alkaloid 

accumulation than shoots or plantlets. Furthermore, somatic embryos are easier 

to handle biotechnologically than more complex structures. Embryogenic strains 

may, therefore, be of special value in further investigations of alkaloid formation 

in N. confusus in vitro. 

Plantlets regenerated from either shoot-clumps or somatic embryos were found 

to produce alkaloids. Table 7.1 shows that only the content of galanthamine 

increased with tissue differentiation. A possible explanation is that the differentiation 

stage could influence the biosynthetic pathway of these alkaloids in a different 

manner. Galanthamine and N-formylnorgalanthamine derive from a para-ortho 

oxidative phenolic coupling, whereas haemanthamine and tazettine result from 

para-para coupling. In general, the more tissue organisation there is, the higher 

the alkaloid content. Plantlets regenerated after 6 months were characterised by a 

well formed bulb and emerging leaves, and seemed to be the best material for 

experiments on galanthamine production. 

Bulb-derived plant material 

Shoots were obtained directly from the bulbs by using the ‘twin-scaling’ technique 

(Hanks and Jones, 1986), and they were classified according to the position of the 

scales in the bulb: internal, central or external. The alkaloid content was analysed 

in the scales initially and in shoots obtained from each kind of scale (Sellés, 1996). 

The different kinds of explants were maintained in the same culture conditions 

for 5 months as previously described (Bergoñón et al., 1996), and then the alkaloids 

were analysed in samples from the regenerated shoots and their respective scales 

(Figure 7.4). In all cases, the major alkaloids were haemanthamine and galanthamine, 

in that order, and the alkaloid profile of the regenerated shoots was 

found to coincide with that of the bulb scales, both qualitatively and quantitatively. 

It is important that all the shoots induced after 5 months are small shoots with 

the capacity to accumulate galanthamine. Therefore, these explants, after being 

mg/g DW 

1.0 

0.8 

0.6 

0.4 

0.2 

0.0 

Outer Middle Inner Average 

Shoots 


A 

1.0 

0.8 

0.6 

0.4 

0.2 

0.0 

Outer Middle Inner Average 

Scales 


Figure 7.4 Alkaloid content in shoots (A) and their respective twin-scales (B) according to 

their position in the bulb (values represent means of three replicates). For 

abbreviations, see Figure 7.1. 

mg/g DW 

B


subcultured and developed in a multiplication medium, also constituted good plant 

material for further experiments on the production of galanthamine type alkaloids. 

EFFECT OF GROWTH REGULATORS ON GROWTH AND 

MORPHOLOGY OF EXPLANTS AND ALKALOID PRODUCTION 

Although the growth of aggregates of both buds and shoots of Narcissus in liquid 

medium has been succesfully achieved without adding any kind of phytohormones 

(Bergoñón et al., 1992), the influence of two kinds of growth regulators has been 

studied: cytokinins (BA and kinetin), which are responsible for both cell division 

and growth of the aerial part of the shoots, and paclobutrazol, an inhibitor of the 

synthesis of gibberellins, which has been previously tested with other bulbous 

plants such as Gladiolus (Steinitz and Lilien-Kipnis, 1989; Steinitz et al., 1991; Ziv, 

1992) and Nerine (Lilien-Kipnis et al., 1992). The main action of paclobutrazol is to 

inhibit the growth of the aerial part of the explants, thus producing wider bulbs. 

Effect of cytokinins 

It has been observed that higher levels of growth regulators are needed in Narcissus 

plants than in other monocots to induce adventitious shoots (Seabrook et al., 1976; 

Seabrook, 1990) and bulblets (Keller, 1993), but their influence in liquid medium 

cultures has not been previously studied. 

Although the addition of cytokinins to the liquid medium is not very advisable, 

especially at high concentrations, because they induce vitrification of the plant 

tissues (Hussey, 1986), this experiment was carried out using seed-derived shootclumps 

with a few (four or five) emergent leaves (Sellés, 1996). The clusters were 

grown for two months in baby-food jars with MS liquid medium supplemented 

with 6% sucrose and cytokinin (BA or kinetin) at concentrations of 0, 1, 3, 5 or 

10 mg/l. Every two weeks, coinciding with the subculturing steps, the alkaloids 

released to the liquid medium were measured. The alkaloids accumulated in the 

tissues were determined at the end of the experiment. In the present chapter, we 

have considered the ‘total production’ of alkaloids as the sum of the alkaloids 

released to the liquid medium during the successive subculturing steps, and those 

accumulated in the tissues at the end of the experiment. 

Benzyladenine 

In general, in the shoot-clumps growing in a liquid medium with cytokinins, the 

aerial part was more developed than the bulb, exhibiting narrow, intensely green 

leaves. Under these culture conditions the percentage of survival was around 90%, 

with the exception of the clusters treated with 10 mg/l of BA, 80% of which became 

vitrified and necrotic. 

Accumulation of alkaloids in the liquid medium 

The results concerning the levels of alkaloids released by the shoot-clumps to the 

liquid medium during the two-month experiment are shown in Figure 7.5. The

222 C. Codina 

µ g/g FW 

µ g/g FW 

30 

25 

20 

15 

10 

5 

0 

25 

20 

15 

10 

5 

0 

N -formylnorgalanthamine 

2 4 6 8 

Time (weeks) 

0mg/l 1mg/l 3mg/l 

5mg/l 10mg/l 

Haemanthamine 

2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

Galanthamine 

production of alkaloids was not very high, and it was more or less constant during 

the experiment. N-formylnorgalanthamine and haemanthamine were the main 

alkaloids released into the liquid medium after two weeks in the case of the 

explants treated with 10 mg/l of BA, and, in general, tazettine was the alkaloid with 

the least tendency to be released into the medium. 

Accumulation of alkaloids in tissue 

Eight weeks after starting the culture, the alkaloid content of the shoot-clumps was 

determined, as well as that of the alkaloids released to the liquid medium (Figure 

7.6). In all cases, the concentration of alkaloids was higher in the tissue than in the 

liquid medium, optimum levels being reached in the shoot-clumps treated with 

3 mg/l of BA. The major alkaloids were haemanthamine, reaching values of up to 

2.17 mg/g DW, and galanthamine, reaching 1.42 mg/g DW. 

µ g/g FW 

µ g/g FW 

25 

20 

15 

10 

5 

0 

25 

20 

15 

10 

5 

0 

2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

Tazettine 

2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

Figure 7.5 Levels of alkaloids released by shoot-clumps treated with BA into the culture 

medium throughout the experiment (bars represent the SD calculated from 

three replicates).

mg/g DW 

mg/g DW 

0.90 

0.75 

0.60 

0.45 

0.30 

0.15 

0.00 

3.50 

3.00 

2.50 

2.00 

1.50 

1.00 

0.50 

0.00 

N-formylnorgalanthamine 

0 1 3 

BA (mg/l) 

5 10 

Haemanthamine 

0 1 3 

BA (mg/l) 

5 10 

ts 

cm 

ts 

cm 


mg/g DW 

mg/g DW 

Galanthamine 

Total production of alkaloids during the experiment 

The levels of alkaloids produced throughout culture are shown in Table 7.2. The 

highest accumulation of alkaloids was observed in the shoot-clumps treated with 

3 mg/l of BA, which also displayed the best growth, followed by those treated with 

1 mg/l of BA. The absence of BA coincided with the lowest accumulation of alkaloids. 

2.50 

2.00 

1.50 

1.00 

0.50 

0.00 

0.35 

0.30 

0.25 

0.20 

0.15 

0.10 

0.05 

0.00 

0 1 3 

BA (mg/l) 

5 10 

Tazettine 

0 1 3 

BA (mg/l) 

5 10 

Figure 7.6 Accumulation of alkaloids in both tissue (shoot-clumps) and liquid medium at 

the end of the experiment with BA. Bars represent the SD calculated from three 

replicates. ts, alkaloids detected in the tissue; cm, alkaloids released to the 

culture medium. 

Table 7.2 Total production of alkaloids (accumulation in both tissue and liquid 

medium) in shoot-clumps grown under different concentrations of BA 

(µg/g FW ± SD calculated from three replicates) a 

BA NFNGAL GAL HAEM TAZ 

0 mg/l 284.2 ± 14.1 475.5 ± 35.1 488.8 ± 39.0 79.7 ± 5.6 

1 mg/l 355.6 ± 19.4 903.7 ± 52.3 1358.5 ± 78.5 157.4 ± 8.2 

3 mg/l 687.4 ± 38.7 1354.2 ± 83.3 2097.5 ± 129.1 248.7 ± 13.2 

5 mg/l 347.1 ± 23.4 572.1 ± 46.9 856.1 ± 77.2 110.8 ± 6.8 

10 mg/l 377.8 ± 14.3 465.7 ± 18.3 673.0 ± 30.1 115.3 ± 6.8 

Note 

a for abbreviations, see Table 7.1. 

ts 

cm 

ts 

cm

224 C. Codina 

Kinetin 

As observed in the experiment with BA, the control shoots developed narrow 

leaves and a well formed bulb, as well as roots. On the contrary, bulbs subjected to 

high doses of kinetin (5 and 10 mg/l) had a necrotic and even vitrified appearance, 

like shoot-clumps treated with the same concentrations of BA. 


The release of alkaloids into the liquid medium of the shoot-clumps treated with 

kinetin was similar to that observed in the clusters grown in media supplemented 

with BA. However, what was notable was the high proportion of the alkaloids 

galanthamine and haemanthamine released into the medium by the explants 

treated with 10 mg/l of kinetin four weeks after starting the culture, which reached 

values of up to 73.6 and 95.6 µg/g fresh weight (FW), respectively (Figure 7.7). 

µg/g FW 

µg/g FW 

25 

20 

15 

10 

5 

0 

120 

100 

80 

60 

40 

20 

0 


2 4 6 

Time (weeks) 


5mg/l 10mg/l 

Haemanthamine 

2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

8 

µg/g FW 

µg/g FW 

100 

25 

20 

15 

10 

80 

60 

40 

20 

5 

0 

0 

Galanthamine 

2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

Tazettine 

2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

Figure 7.7 Levels of alkaloids released by shoot-clumps treated with kinetin into the culture 

medium throughout the experiment (bars represent the SD calculated from 

three replicates).



Unlike BA, kinetin, especially at high concentrations, inhibited alkaloid production 

in the shoot tissues (Figure 7.8), as has also been described for cultures of Scopolia 

maxima (Mantell and Smith, 1983). Thus, the maximum levels of alkaloids 

were found in the control shoot-clumps, with the exception of tazettine, whose 

maximum level was found in the shoot-clumps treated with 3 mg/l of kinetin. This 

growth regulator seems to have more effect on the release of alkaloids to the 

medium than on their accumulation in the tissues. 


The total content of alkaloids (those accumulated in tissue and those released into 

the medium) produced by the shoot-clumps treated with different concentrations 

of kinetin during the two-month experiment is shown in Table 7.3. Kinetin 

affected the alkaloid production in a negative way, so the highest levels of these 

compounds, especially of haemanthamine and galanthamine, were found in the 

control shoots. 

mg/g DW 

mg/g DW 

0.35 

0.30 

0.25 

0.20 

0.15 

0.10 

0.05 

0.00 

0.40 

0.35 

0.30 

0.25 

0.20 

0.15 

0.10 

0.05 

0.00 


0 1 3 

Kin (mg/l) 

5 10 

Haemanthamine 

0 1 3 

Kin (mg/l) 

5 10 

ts 

cm 

ts 

cm 

mg/g DW 

mg/g DW 

0.50 

0.40 

0.30 

0.20 

0.10 

0.00 

0.35 

0.30 

0.25 

0.20 

0.15 

0.10 

0.05 

0.00 

Galanthamine 

0 1 3 

Kin (mg/l) 

5 10 

Tazettine 

0 1 3 

Kin (mg/l) 

5 10 


the end of the experiment with kinetin (Kin) (bars represent the SD calculated 

from three replicates). ts, alkaloids detected in the tissue; cm, alkaloids released 

to the culture medium. 

ts 

cm 

ts 

cm

226 C. Codina 


medium) in shoot-clumps grown under different concentrations of 

kinetin (µg/g FW ± SD calculated from three replicates) a 

Kinetin NFNGAL GAL HAEM TAZ 

0 mg/l 274.9 ± 9.5 358.9 ± 26.1 368.9 ± 55.2 139.2 ± 21.9 

1 mg/l 209.1 ± 7.2 179.1 ± 15.1 173.0 ± 22.6 147.8 ± 4.3 

3 mg/l 144.1 ± 6.6 182.2 ± 13.3 142.9 ± 9.0 208.6 ± 3.9 

5mg/l 95.1± 8.8 159.2 ± 19.9 103.4 ± 8.2 39.3 ± 1.9 

10 mg/l 98.2 ± 8.3 225.8 ± 37.0 188.7 ± 38.1 32.8 ± 5.2 

Note 


Comparing the effects of these cytokinins on explant development and alkaloid 

production, one can observe that shoot-clumps treated with BA showed a better 

development than those treated with kinetin. The best treatment for the multiplication 

of the explants was the concentration of 3 mg/l of BA, and the controls, 

with a lower number of leaves, accumulated a higher amount of dry matter. In 

addition, the degree of necrosis in the experiment with kinetin was higher than in 

the other experiments, which could be related to a higher release of alkaloids by 

the shoot-clumps. In comparison with the control explants, the alkaloid content, 

in general, was strongly influenced by the different concentrations of BA, galanthamine 

and haemanthamine being the predominant alkaloids produced. 

The results of this experiment reveal that Narcissus confusus is not especially 

susceptible to vitrification, which is usually found when tissues are cultivated in a 

liquid medium and, in addition, under high concentrations of cytokinins (Hussey, 

1986). This could be due to the fact that the subcultures were systematically performed 

every two weeks, instead of every five weeks as described for other species 

of this genus (Chow et al., 1993). This higher frequency of subculturing could have 

avoided the excessive accumulation of ethylene inside the culture flasks, which is 

also involved in the vitrification process (Hussey, 1986). 

Effect of paclobutrazol 

Paclobutrazol, an inhibitor of gibberellin biosynthesis, has been used in liquid 

cultures with other plants belonging to the genera Gladiolus (Steinitz and Lilien- 

Kipnis, 1989), Nerine (Lilien-Kipnis et al., 1992), Colchicum (Ellington, 1998), Solanum 

(Simko, 1994), Nephrolepis and Philodendron (Ziv, 1992), and its influence on both 

the proliferation of meristematic shoots and the lateral growth of explants has 

been observed. 

The effect of paclobutrazol has been examined on explants with different levels 

of development: bulblets with emerging leaves and well formed bulbs, and shootclumps. 

The explants were obtained as previously described (Sellés, 1996), and 

cultured in baby-food jars with MS liquid medium supplemented with 6% sucrose 

and paclobutrazol at concentrations of 0, 1, 3, 5 and 10 mg/l. They were cultivated 

for two months at 25 ± 1 °C under a photoperiod of 16 h, and shaken at 110 rpm. 

The concentrations of the alkaloids accumulated in tissue were determined 8 weeks 

after starting the experiment, whereas the release of alkaloids to the liquid medium 

was measured every two weeks, coinciding with the subculturing steps.


Bulblets 

As a consequence of the effect of paclobutrazol, the shoots formed during the 

8-week experiment were characterised by the absence of leaves and roots, and 

showed an especially well-developed bulb. 


The release of alkaloids into the liquid medium of the explants treated with 

paclobutrazol decreased during the experiment (Figure 7.9). Independently of 

the paclobutrazol concentration, in all the experiments the main alkaloids released 

to the medium were haemanthamine and galanthamine, whose levels reached maximum 

values of 71.3 and 35.8 µg/g FW, respectively. Tazettine was not excreted 

into the liquid medium in the last sample. The treatment that promoted the highest 

release of alkaloids to the medium was 5 mg/l of paclobutrazol. 

µg/g FW 

µg/g FW 

30 

25 

20 

15 

10 

5 

0 

80 

70 

60 

50 

40 

30 

20 

10 

0 


2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

Haemanthamine 

2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

µg/g FW 

µg/g FW 

45 

40 

35 

30 

25 

20 

15 

10 

5 

0 

25 

20 

15 

10 

5 

0 

Galanthamine 

2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

Tazettine 

2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

Figure 7.9 Levels of alkaloids released by bulblets treated with paclobutrazol into the culture 

medium throughout the experiment (bars represent the SD calculated 

from three replicates).

228 C. Codina 


The alkaloid content in tissues at the end of the experiment was higher than that 

of the culture medium, haemanthamine and galanthamine being the major alkaloids 

(Figure 7.10). The control explants (grown without paclobutrazol) accumulated 

the highest concentration of alkaloids. 


The final alkaloid content (alkaloids accumulated in tissue and those released to 

the liquid medium) at the end of the experiment, after a two-month period, is 

shown in Table 7.4. The control explants were the most productive, showing a 

good appearance with narrow leaves and well developed roots. 

Shoot-clumps 

In this experiment, some necrosis and vitrification problems appeared from the 

fourth sample onwards in all the explants treated with paclobutrazol. These problems 

were less notable in the control shoot-clumps and those treated with the lowest 

mg/g DW 

mg/g DW 

0.14 

0.12 

0.10 

0.08 

0.06 

0.04 

0.02 

0.00 

0.60 

0.50 

0.40 

0.30 

0.20 

0.10 

0.00 


0 1 3 5 10 

Paclobutrazol (mg/l) 

Haemanthamine 

0 1 3 5 10 


ts 

cm 

ts 

cm 

mg/g DW 

mg/g DW 

0.50 

0.40 

0.30 

0.20 

0.10 

0.00 

0.12 

0.10 

0.08 

0.06 

0.04 

0.02 

0.00 

Galanthamine 

0 1 3 5 10 


Tazettine 

0 1 3 5 10 


Figure 7.10 Accumulation of alkaloids in both tissue (bulblets) and liquid medium at the 

end of the experiment with paclobutrazol (bars represent the SD calculated 



ts 

cm 

ts 

cm

µg/g FW 

µg/g FW 


Table 7.4 Total production of alkaloids (accumulation in both tissue and 

liquid medium) in bulblets grown under different concentrations of 

paclobutrazol (µg/g FW ± SD calculated from three replicates) a 

Paclobutrazol NFNGAL GAL HAEM TAZ 

0 mg/l 56.6 ± 5.6 118.8 ± 4.2 170.9 ± 4.8 23.8 ± 3.4 

1 mg/l 52.8 ± 0.9 99.1 ± 1.2 91.1 ± 6.8 16.6 ± 1.4 

3 mg/l 43.4 ± 4.4 60.3 ± 4.9 53.2 ± 2.5 11.9 ± 0.8 

5 mg/l 56.6 ± 3.8 99.6 ± 2.2 123.0 ± 3.6 38.4 ± 2.7 

10 mg/l 56.5 ± 0.4 72.9 ± 4.4 124.8 ± 3.4 26.8 ± 4.2 

Note 


300 

250 

200 

150 

100 

50 

0 

350 

300 

250 

200 

150 

100 

50 

0 


2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

Haemanthamine 

2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

µg/g FW 

µg/g FW 

Galanthamine 

concentration of the growth regulator. The control shoot-clumps had a good 

appearance, each one showing about eight to nine shoots at the end of the experiment. 

The lower growth index of the explants treated with the highest doses of 

paclobutrazol agrees with observations in Gladiolus (Ziv, 1992). 

700 

600 

500 

400 

300 

200 

100 

0 

160 

140 

120 

100 

80 

60 

40 

20 

0 

2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

Tazettine 

2 4 6 8 

Time (weeks) 


5mg/l 10mg/l 

Figure 7.11 Levels of alkaloids released by shoot-clumps treated with paclobutrazol into the 

culture medium throughout the experiment (bars represent the SD calculated 

from three replicates).

230 C. Codina 


Although the release of alkaloids into the medium increased until the fourth week 

of culture, from then on, and coinciding with the necrosis and vitrification of the 

tissues, excretion decreased drastically (Figure 7.11). As in the bulblets, the main 

alkaloids released to the medium were galanthamine and haemanthamine, reaching 

values of 404.2 and 295.5 µg/g FW, respectively. In general, the levels of alkaloids 

released by the shoot-clumps into the liquid medium were higher than those 

released by the bulblets. 


In the control shoot-clumps, or in those treated with low doses of paclobutrazol, 

the alkaloids were accumulated mainly in the tissue, whereas at concentrations of 

5 mg/l or more of this growth regulator the alkaloids were mainly released to the 

culture medium (Figure 7.12). This fact could be related to the degree of vitrification, 

which was proportional to the concentration of paclobutrazol. 

mg/g DW 

mg/g DW 

0.70 

0.60 

0.50 

0.40 

0.30 

0.20 

0.10 

0.00 

0.80 

0.70 

0.60 

0.50 

0.40 

0.30 

0.20 

0.10 

0.00 


0 1 3 5 10 


Haemanthamine 

0 1 3 5 10 


ts 

cm 

ts 

cm 

mg/g DW 

mg/g DW 

0.70 

0.60 

0.50 

0.40 

0.30 

0.20 

0.10 

0.00 

0.14 

0.12 

0.10 

0.08 

0.06 

0.04 

0.02 

0.00 

Galanthamine 

0 1 3 5 10 


Tazettine 

0 1 3 5 10 



the end of the experiment with paclobutrazol (bars represent the SD calculated 



ts 

cm 

ts 

cm



medium) in shoot-clumps grown under different concentrations of 

paclobutrazol (µg/g FW ± SD calculated from three replicates) a 

Paclobutrazol NFNGAL GAL HAEM TAZ 

0 mg/l 298.7 ± 12.4 329.4 ± 77.1 418.4 ± 43.4 149.2 ± 24.1 

1 mg/l 159.5 ± 31.2 208.4 ± 72.4 177.9 ± 33.4 81.1 ± 15.0 

3 mg/l 198.3 ± 17.1 268.4 ± 102.2 231.8 ± 16.2 109.9 ± 3.4 

5 mg/l 208.6 ± 6.6 307.4 ± 108.7 243.5 ± 106.1 119.2 ± 4.6 

10 mg/l 320.5 ± 25.0 465.1 ± 221.1 346.0 ± 60.2 177.4 ± 3.8 

Note 



The total production of alkaloids in the shoot-clumps treated with 1, 3 or 5 mg/l of 

paclobutrazol was lower than in the controls (Table 7.5). Nevertheless, the highest 

production took place in those treated with 10 mg/l of paclobutrazol. Galanthamine 

was the main alkaloid produced under the influence of paclobutrazol, 

whereas the control shoot-clumps accumulated mainly haemanthamine. 

By comparing the effect of paclobutrazol on both the morphology of the 

explants and the production of alkaloids, one can observe that the bulblets were 

better developed than the shoot-clumps, as the latter became necrotic. In general, 

growth was higher in the explants (bulblets and shoot-clumps) treated with 

paclobutrazol, as was also observed in Gladiolus (Ziv, 1992), although the differences 

among the different treatments were small. In both kinds of explants, the presence 

of paclobutrazol in the medium, promoted the development of adventitious 

shoots and, in general, inhibited the synthesis of alkaloids in comparison with the 

control explants. The excretion of alkaloids into the liquid medium was higher in 

shoot-clumps than in bulblets. 

EFFECT OF SUCROSE CONCENTRATION ON GROWTH 

AND PRODUCTION OF ALKALOIDS 

The importance of sucrose, a source of carbon essential for the growth and development 

of bulbs, has previously been demonstrated in plants belonging to the 

genera Allium (Keller, 1993), Lilium (Takayama and Misawa, 1979), Tulipa (Taeb 

and Alderson, 1990), Narcissus (Chow et al., 1992) and Gladiolus (Steinitz et al., 

1991). With the exception of Gladiolus, these experiments were carried out in solid 

agar media. The present experiments were performed in liquid medium with 

shoot-clumps derived from both bulbs and seeds of Narcissus confusus, which were 

treated with different concentrations of sucrose, 30, 60, 90, 120, 150 and 180 g/l 

(Sellés et al., 1997b). The experiment took place over two weeks, as the secondary 

metabolites are usually formed at the end of the growth period, when the nutrients 

of the medium become limiting.

232 C. Codina 

Effect on growth and morphology of the explants 

The different origin of the shoot-clumps used in this experiment (bulbs or seeds) 

did not seem to influence significantly the growth of the explants, although the 

shoot-clumps derived from bulbs were slightly bigger. In both cases, the clusters 

formed in the treatment with 90 g/l of sucrose exhibited the best growth index of 

weight, showing a well formed bulb and emerging intensely green leaves. The 

shoot-clumps treated with concentrations of sucrose higher than 90 g/l showed 

pale leaves, with a tendency to vitrification. In addition, the tissue of the base of 

the bulb was yellowish, sometimes showing necrotic areas, and the emergence of 

leaves was less vigorous. Apparently, high concentrations of sucrose (≥ 150 g/l) 

became toxic to the explants derived from both strains of shoot-clumps (coming 

from bulbs or seeds), and caused shoot dormancy. 

Effect on alkaloid production 

Although the two strains showed a similar growth and morphology, they exhibited 

different behaviour in relation to the production of alkaloids. 

Bulb-derived shoot-clumps 

The effect of sucrose concentrations on the production of alkaloids in bulb-derived 

shoot-clumps is shown in Figure 7.13. The main alkaloid was haemanthamine, 

with a value of 0.87 mg/g DW in the treatment with the lowest sucrose concentration 

(3%), whereas the minor alkaloid was tazettine, with a value of 0.15 mg/g 

DW in the same treatment. The maximum accumulation of galanthamine took 

place in the treatments with the highest and lowest doses of sucrose (3 and 18%), 

reaching practically the same value, 50 mg/g DW. The maximum production 

of N-formylnorgalanthamine occurred in the explants treated with 30 g/l of 

sucrose. 

In general, the qualitative profile of the alkaloids in the different treatments 

with sucrose was very similar. From the quantitative point of view, however, an 

increase in the source of carbon led to a corresponding decrease in the production 

of alkaloids, the minimum value thus being obtained in the treatment with 90 g/l of 

sucrose. At higher concentrations of sucrose, the production of alkaloids increased, 

but without reaching the maximum values observed in the treatment with 30 g/l of 

sucrose. In all cases, the alkaloids were mainly accumulated in the tissue, a very 

small proportion being released to the culture medium (Figure 7.13). 

Seed-derived shoot-clumps 

The production of alkaloids in the seed-derived shoot-clumps and treated 

with different doses of sucrose are shown in Figure 7.14. In this case, one can 

observe that the accumulation of galanthamine type alkaloids was higher in 

the medium culture than in the tissue, except in the treatments with 90 and 

120 g/l of sucrose. In the case of the haemanthamine type alkaloids, the percentage 

accumulated in the tissue was generally higher than that released to 

the liquid medium.

mg/g DW 

mg/g DW 

0.40 

0.35 

0.30 

0.25 

0.20 

0.15 

0.10 

0.05 

0.00 

1.20 

1.00 

0.80 

0.60 

0.40 

0.20 

0.00 


30 60 90 120 150 180 

Sucrose (g/l) 

Haemanthamine 

30 60 90 120 150 180 

Sucrose (g/l) 

ts 

cm 

ts 

cm 

mg/g DW 

mg/g DW 


Galanthamine 

As in the case of the shoot-clumps derived from twin-scales, the maximum accumulation 

of alkaloids took place at low concentrations of sucrose, and it decreased 

progressively as the source of carbon increased, especially in the case of the galanthamine 

type alkaloids. Nonetheless, the alkaloid profile is not exactly the same as 

that observed in the bulb-derived shoot-clumps. In this case, the major alkaloids 

were N-formylnorgalanthamine and galanthamine, reaching a maximum accumulation 

in the treatment with 30 g/l of sucrose. 

Comparing the effect of the sucrose on alkaloid production in the two kinds of 

explants used in this experiment, it is observed that generally the shoot-clumps 

derived from seeds produced a higher amount of alkaloids than those derived 

from bulbs (Table 7.6). The highest content of galanthamine was 0.61 mg/g DW in 

the seed-derived shoot-clumps treated with 3% of sucrose. Likewise, the release of 

alkaloids to the culture medium was also higher in the seed-derived shoot-clumps 

than in those derived from bulbs. Although the causes are unknown, there are 

several factors that might be influential, for instance, the initial physiological state 

of the inoculum. 

The results obtained in the two strains show that the production of alkaloids 

does not seem to be directly related to the growth of the explants, as the maximum 

0.70 

0.60 

0.50 

0.40 

0.30 

0.20 

0.10 

0.00 

0.25 

0.20 

0.15 

0.10 

0.05 

0.00 

30 60 90 120 150 180 

Sucrose (g/l) 

Tazettine 

30 60 90 120 150 180 

Sucrose (g/l) 

Figure 7.13 Alkaloid production in bulb-derived shoot-clumps grown under different 

sucrose concentrations. Bars represent the SD calculated from three replicates. 

ts, alkaloids detected in the tissue; cm, alkaloids released to the culture medium. 

ts 

cm 

ts 

cm

234 C. Codina 

mg/g DW 

mg/g DW 

0.90 

0.75 

0.60 

0.45 

0.30 

0.15 

0.00 

0.60 

0.50 

0.40 

0.30 

0.20 

0.10 

0.00 


30 60 90 120 150 180 

Sucrose (g/l) 

Haemanthamine 

30 60 90 120 150 180 

Sucrose (g/l) 

ts 

cm 

ts 

cm 

mg/g DW 

mg/g DW 

Galanthamine 

growth index (at 90 g/l of sucrose) does not coincide with the maximum concentration 

of alkaloids (at 30 g/l of sucrose). Although both strains showed similar 

growth, the seed-derived shoot-clumps constituted a better kind of plant material 

for alkaloid production than those obtained from adult bulbs. 

0.90 

0.75 

0.60 

0.45 

0.30 

0.15 

0.00 

0.70 

0.60 

0.50 

0.40 

0.30 

0.20 

0.10 

0.00 

30 60 90 120 150 180 

Sucrose (g/l) 

Tazettine 

30 60 90 120 150 180 

Sucrose (g/l) 

Figure 7.14 Alkaloid production in seed-derived shoot-clumps grown under different 

sucrose concentrations. Bars represent the SD calculated from three replicates. 

ts, alkaloids detected in the tissue; cm, alkaloids released to the culture medium. 

Table 7.6 Effect of sucrose on alkaloid production (accumulation in both tissue and liquid 

medium) in shoot-clumps obtained from bulbs and seeds (values are % DW) a 

Sucrose (g/l) Bulb-derived shoot-clumps Seed-derived shoot-clumps 

Note 


NFNGAL GAL HAEM TAZ NFNGAL GAL HAEM TAZ 

30 0.028 0.028 0.082 0.015 0.065 0.061 0.057 0.037 

60 0.018 0.021 0.041 0.009 0.048 0.040 0.031 0.021 

90 0.008 0.021 0.022 0.005 0.045 0.030 0.041 0.043 

120 0.014 0.021 0.043 0.008 0.020 0.018 0.025 0.025 

150 0.009 0.022 0.032 0.004 0.021 0.023 0.023 0.012 

180 0.024 0.047 0.061 0.009 0.012 0.010 0.011 0.008 

ts 

cm 

ts 

cm


EFFECT OF CULTURE VESSEL SIZE ON GROWTH 

AND ALKALOID PRODUCTION 

After observing that the best treatment for the growth of the explants in liquid 

medium was that of 90 g/l of sucrose, an experiment to produce alkaloids during a 

longer period of time (two months) was designed. Unlike the previous experiments, 

this was performed with small plantlets obtained from a strain of seedderived 

organogenic callus, and two sizes of culture flasks (250 and 500 ml) were 

used to check the possible influence of this factor on the growth and morphology 

of explants and alkaloid production (Sellés, 1996). The selected plantlets showed a 

well formed bulb (ca. 0.9–1.3 cm), with emerging leaves (2–3 cm in length) and 

small shoots at the base of the bulb. They were cultured in a liquid MS medium 

supplemented with 90 g/l of sucrose. The 250 ml and 500 ml erlenmeyer flasks 

contained 50 ml and 100 ml of liquid medium, respectively. The samples were 

analysed every two weeks, coinciding with subculturing, and the morphology of 

the plantlets and the accumulation of alkaloids were determined. 

Effect on growth and morphology of the explants 

In general, plantlets grown in both sizes of vessels showed well formed bulbs, with 

leaves up to 8–10 cm in length. The number of shoots developed by the end of the 

experiment normally coincided with the number of small buds of the initial 

explant, and they showed good development with apical dominance of the central 

shoot. Some vitrification and necrosis were observed during the experiment in 

about 7% of the explants. These problems were more notable in the 500 ml flasks, 

probably because the explants were practically submerged in the liquid culture in 

these vessels which prejudiced foliar development and promoted the growth of 

bulbs. 

Effect on alkaloid production 

In the first sample, the excretion of alkaloids to the culture medium was not determined, 

only accumulation in the tissue. 

250 ml flasks experiment 

Figure 7.15 shows the results of the alkaloids produced by the plantlets cultured in 

the 250 ml flasks. In general, no significant fluctuations in the alkaloid production 

were observed during liquid culture, although the highest levels were observed in 

the first subculturing step. The main alkaloids were found to be galanthamine and 

haemanthamine, which reached total (tissue and liquid culture) levels of 1.54 and 

1.51 mg/g DW, respectively. The highest accumulation of N-formylnorgalanthamine 

took place four weeks after starting the experiment, reaching 1.01 mg/g DW. The 

total levels of tazettine were found to be more or less constant throughout the 

experiment, but they were always lower than those accumulated by the explants 

before starting the assay.

236 C. Codina 

mg/g DW 

mg/g DW 

1.4 

1.2 

1.0 

0.8 

0.6 

0.4 

0.2 

0.0 

1.6 

1.4 

1.2 

1.0 

0.8 

0.6 

0.4 

0.2 

0.0 


0 2 4 6 8 

Time (weeks) 

Haemanthamine 

0 2 4 6 8 

Time (weeks) 

ts 

cm 

ts 

cm 

Galanthamine 

In general, the accumulation of alkaloids was always higher in the tissue than in 

the culture medium, except in the case of the galanthamine type alkaloids, 

especially four weeks after starting the experiment in the liquid medium culture. 

From the beginning, N-formylnorgalanthamine was the alkaloid most susceptible 

to being released into the culture medium. In all the cases, the maximum concentration 

of alkaloids in tissue occurred in the final step of the experiment. This was 

probably due to the fact that the plantlets kept in this kind of flask were close to 

the limit of exponential growth, when the accumulation of secondary plant metabolites 

is usually at its highest (Maldonado-Mendoza et al., 1993). 

500 ml flasks experiment 

As in the former experiment, plantlets cultivated in 500 ml flasks produced 

galanthamine and haemanthamine as the major alkaloids, also showing maximum 

concentration two weeks after starting the experiment, with total (tissue and 

culture medium) values of 1.24 and 1.32 mg/g DW, respectively (Figure 7.16). In 

addition, N-formylnorgalanthamine and tazettine also reached maximum levels in 

the same period, 1.18 and 0.38 mg/g DW, respectively. 

1.4 

1.2 

1.0 

0.8 

0.6 

0.4 

0.2 

0.0 

1.0 

0.8 

0.6 

0.4 

0.2 

0.0 

0 2 4 6 8 

Time (weeks) 

Tazettine 

0 2 4 6 8 

Time (weeks) 

Figure 7.15 Alkaloid production by plantlets grown in 250 ml flasks. Bars represent 

the SD calculated from three replicates. ts, alkaloids detected in the tissue; 

cm, alkaloids released to the culture medium. 

mg/g DW 

mg/g DW 

ts 

cm 

ts 

cm

mg/g DW 

mg/g DW 

1.4 

1.2 

1.0 

0.8 

0.6 

0.4 

0.2 

0.0 

1.8 

1.5 

1.2 

0.9 

0.6 

0.3 

0.0 


0 2 4 6 8 

Time (weeks) 

Haemanthamine 

0 2 4 6 8 

Time (weeks) 

ts 

cm 

ts 

cm 


Galanthamine 

As in the 250 ml flask experiment, the main alkaloid released to the medium 

culture in the 500 ml flasks was found to be N-formylnorgalanthamine, especially 

in the first subculturing steps. In general, there was a higher accumulation of 

alkaloids in the tissue than in the liquid medium. Nonetheless, the maximum concentration 

of alkaloids in this case did not take place at the end of the experiment, 

probably because the plantlets still had room to grow in the bigger flasks, thus 

being far from their maximum potential growth. 

Comparing the effect of the size of the culture vessels on the alkaloid production 

in N. confusus plantlets one can first observe that the alkaloid profile of the 

explants is not always the same (Figure 7.17). Thus, in 250 ml flasks the total concentration 

of alkaloids stayed more or less constant throughout the experiment, 

with minor fluctuations. The main alkaloids were found to be galanthamine and 

haemanthamine, showing the highest levels two weeks after starting the culture. 

The accumulation of N-formylnorgalanthamine and tazettine was more or less 

constant throughout the experiment, with the exception of the minimum levels 

corresponding to 6 weeks of culture. The plantlets grown in 500 ml flasks exhibited 

a similar behaviour, and except for tazettine, all the alkaloids reached maximum and 

minimum concentrations two and six weeks after starting the culture, respectively. 

mg/g DW 

mg/g DW 

1.4 

1.2 

1.0 

0.8 

0.6 

0.4 

0.2 

0.0 

1.0 

0.8 

0.6 

0.4 

0.2 

0.0 

0 2 4 6 8 

Time (weeks) 

Tazettine 

0 2 4 6 8 

Time (weeks) 

Figure 7.16 Alkaloid production by plantlets grown in 500 ml flasks. Bars represent the 

SD calculated from three replicates. ts, alkaloids detected in the tissue; cm, 

alkaloids released to the culture medium. 

ts 

cm 

ts 

cm

238 C. Codina 

mg/g DW 

mg/g DW 

1.5 

1.0 

0.5 

0.0 

1.5 

1.0 

0.5 

0.0 

The maximum concentration of alkaloids in both sizes of flasks was observed in 

the first subculture. This could be due to the fact that the explants have to adapt to 

new environmental conditions at an early stage, when transferred from a solid 

medium with growth regulators to a liquid one without phytohormones and with 

a higher than normal concentration of sucrose. This stressful situation could be 

responsible for a higher synthesis of alkaloids. 

In general, the accumulation of alkaloids was higher in 250 ml than 500 ml 

flasks, although the maximum levels were similar in both vessels. From a morphological 

point of view, the plantlets obtained at the end of the experiment appeared 

similar, although the leaf development of the explants cultured in 500 ml flasks 

was slightly poorer. As the shaking speed was the same in both cases, the larger 

flasks had more atmospheric oxygen and, therefore, better aerated plantlets, producing 

bulblets that were less stressed and with lower levels of alkaloids. 

CONCLUSIONS 


0 2 4 6 8 

Time (weeks) 

Haemanthamine 

0 2 4 6 8 

Time (weeks) 

500 ml 

250 ml 

500 ml 

250 ml 

The culture of shoot-clumps of Narcissus confusus in liquid medium has been found 

to be a good experimental system for the production of galanthamine and related 

alkaloids. The culture of this kind of explant is improved by the addition of the 

cytokinins BA or kinetin at low concentrations (1–3 mg/l). The accumulation of 

galanthamine in tissues is higher in shoot-clumps treated with BA, whereas its 

release into the culture medium takes place more when kinetin is present. Another 

growth regulator, paclobutrazol, affected the two kinds of explants (bulblets and 

mg/g DW mg/g DW 

1.5 

1.0 

0.5 

0.0 

1.5 

1.0 

0.5 

0.0 

Galanthamine 

0 2 4 6 8 

Time (weeks) 

Tazettine 

0 2 4 6 8 

Time (weeks) 

500 ml 

250 ml 

500 ml 

250 ml 

Figure 7.17 Comparison between the total production of alkaloids (detected in the 

tissue and released to the liquid medium) by plantlets grown in two different 

flask sizes throughout the experiment.


shoot-clumps) in different ways, although it did not contribute to an increase of 

alkaloid accumulation. Sucrose is required for both the growth of the explants and 

alkaloid production. Nonetheless, independently of the origin of the explant used, 

bulbs or seeds, the optimal concentration of sucrose to obtain biomass (90 g/l) does 

not coincide with the best conditions to induce higher alkaloid accumulation (30 g/l). 

In general, the culture of shoot-clumps in liquid medium has been found to be 

stable throughout the experiments (up to two months), the galanthamine-type 

alkaloids being released into the culture medium in a higher proportion, which 

facilitates their extraction. Obviously, apart from these culture conditions, other 

parameters involved in the biosynthesis of galanthamine, as well as technological 

conditions, have still to be studied before considering the possibility of scaling-up 

production using biotechnological methods. 

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8 Narcissus and other Amaryllidaceae 

as sources of galanthamine 

O.A. Cherkasov and O.N. Tolkachev 

INTRODUCTION 

The Amaryllidaceae family contains 65 genera and over 860 species. Amaryllidaceae 

species are widely distributed in the tropical, sub-tropical and temperate 

zones of both hemispheres, and a vast diversity of species is characteristic of 

the flora of the South African Cape region, South and Central America and the 

Mediterranean coasts. They possess highly decorative characteristics, and have 

been used from ancient times in floriculture and medicine (Artyushenko, 1970; 

Khamidkhodzhaev, 1984). 

Amaryllid plants are very important as ornamentals. They are popular for growing 

in parks and gardens, as farm crops and as indoor and greenhouse plants for 

winter and spring colour (Artyushenko, 1963; Tsytsin, 1960; Petrov, 1975). 

All Amaryllidaceae are rich in alkaloids, the principal biologically active components 

of plant extracts. Today over 200 alkaloids have been isolated from plants 

of this family. Incidents of animals being poisoned by Amaryllidaceae, due to the 

alkaloids contained, have been recorded in the literature: poisonous examples 

include Galanthus and Leucojum species (Gusynin, 1955). The presence of alkaloids 

in plants is believed to be a protective adaptation, which in Amaryllidaceae is 

connected with the seasonal cycle of development, many species growing in early 

spring when other genera are only just starting to grow. 

Amaryllidaceae have attracted the attention of scientists as the sources of novel 

compounds, including potentially valuable alkaloids of medicinal importance. The 

chemistry of Amaryllidaceae alkaloids has been reviewed in several reviews and 

monographs (Cook and Loudon, 1952; Cherkasov, 1977; Döpke, 1976, 1978; 

Wildman, 1960, 1968; Fuganti, 1975; Grundon, 1984, 1985, 1987; Jeffs, 1990; 

Lewis, 1990, 1997; Medvedeva et al., 1994; Abduazimov, 1993; Polt, 1996; Ruan, 

1988; Yunusov, 1981). The absolute configuration and ring conformation of 

(–)-galanthamine have recently been determined by crystallographic methods 

(Peeters et al., 1997). 

Galanthamine (Figure 8.1), also known as Galanthin and Nivaline Jilkon (Negwer, 

1978), is the most important pharmaceutically active compound among the 

Amaryllidaceae alkaloids, and it is used in medicine as the hydrobromide. It is 

an anticholinesterase agent of low toxicity used in medicine for the treatment of 

myasthenia, myopathy, neuritis, residual phenomena after poliomyelitis anterior 

acuta (infantile paralysis), psychogenic ‘spinal impotence’, spastic pareses and 

progressive muscular dystrophy, and as the antagonist of muscular relaxants in

MeO 

MeO 

HO 

O 

O 

O 

NMe 

Galanthamine 

HO 

HO 

H 

H 

Lycorine 

Fortucine 

OH 

OH 

N 

H 

N 

OH 

Figure 8.1 Formulae of galanthamine, lycorine and fortucine. 

Sources of galanthamine 243 

the case of surgical interventions (Klyuev and Babayan, 1979; Kovanev et al., 

1967; Krylov, 1999; Mashkovsky and Kruglova-L’vova, 1951; Mashkovsky, 1955, 

1984; Saev and Tenev, 1963; Sokolov and Zamotaev, 1989; Turova and Sapozhnikova, 

1982). It counteracts the sedative, hypnotic or respiratory effects of 

benzazepines and is used for treatment of schizophrenia (Snorrason, 1996). 

Galanthamine is a selective acetylcholinesterase inhibitor which was recently clinically 

trialled for the treatment of Alzheimer’s disease (Fulton and Benfield, 1996; 

Selles et al., 1997b). The biogenetic precursor of galanthamine, (±)-narwedine, has 

been studied as a respiratory stimulator (Bazhenova et al., 1972). Narwedine was 

shown to inhibit the action of narcotics and hypnotics, and to increase the effect of 

analgesics. 

Lycorine (Figure 8.1), also known as Narcissin and Galanthidin (Negwer, 1978), 

and dihydrolycorine have antiarrhythmic action (Aliev, 1972), and lycorine hydrochloride 

is used in medicine in Russia as a strong broncholytic (Klyuev and 

Babayan, 1979). Some alkaloids of this group possess an inhibitory activity on 

herpes simplex virus (Renard-Nozaki et al., 1989). The pharmacological activity 

of Amaryllidaceae alkaloids has been reviewed by Ruan (1988) and Evidente et al. 

(1986).

244 O.A. Cherkasov and O.N. Tolkachev 

A number of galanthamine derivatives have been obtained. A study of structure–activity 

relationships revealed potent N-methyl-galanthamine hydroxide, 

which is more than 20 times as active as galanthamine in anticholinesterase activity 

(Abdusamatov, 1968, 1972). Three-dimentional quantitative structure–activity 

relationship studies of galanthamine and its analogues have been carried out. 

From the calculation of the lowest energy conformations, it was shown that the 

dominant factor was the steric effect, whereas the electrostatic effect of the large 

substituent decreased activity (Luo et al., 1995). 

SOURCES OF GALANTHAMINE AND LYCORINE USED IN MEDICINE 

Galanthamine and lycorine have so far attracted most interest for their medical 

applications. Galanthamine was isolated for the first time from Galanthus woronowii 

Losinsk. by Proskurnina and Yakovleva (1952, 1955), and was afterwards found in 

many other species of Amaryllidaceae, including Leucojum aestivum L., Ungernia 

victoris Vved. and Pancratium and Sternbergia species (Abdusamatov, 1972; Boit, 

1961; Cherkasov, 1975, 1976, 1977; Cherkasov et al., 1984a,b, 1985, 1986, 1988, 

1989; Cherkasov and Tolkachev, 1996; Gorbunova et al., 1978; Maisuradze et al., 

1984, 1985; Sadykov and Khodzhimatov, 1988). On a commercial scale, galanthamine 

has been produced from Galanthus nivalis L. in Bulgaria under the name 

‘Nivalin’. In the former Soviet Union galanthamine was produced from leaves of 

G. woronowii in Russia and from leaves of Ungernia victoris in Uzbekistan (Abdusamatov, 

1968, 1972; Cherkasov et al., 1984a; Khamidkhodzhaev, 1967, 1977, 1982). 

U. victoris is an endemic plant growing widely in Tajikistan in the Gissar mountain 

region, the natural area of which is limited (Cherkasov et al., 1984a). Leaves of the 

plant contain 0.05–0.15% galanthamine, while its content in bulbs varies from 

0.05% up to 0.40–0.60% in some populations. The limited supply of U. victoris has 

been exhausted by commercial harvesting. Attempts at the cultivation of Ungernia 

were unsuccessful because of its slow regeneration: it would require about 45–50 

years to restore the former production from small bulbs or from seeds. As the natural 

supply of the plant material does not satisfy the total demands of the pharmaceutical 

industry, a search for alternative natural sources of galanthamine is of 

great importance (Cherkasov, 1977; Cherkasov et al., 1984a; Khamidkhodzhaev, 

1967, 1984). 

Measurement of the galanthamine levels in Amaryllidaceae has been carried out 

by a number of groups in Russia (e.g., Patudin et al., 1978; Gorbunova et al., 1978; 

Sadykov and Khodzhimatov, 1988). The concentration of galanthamine in 

Amaryllidaceae has been found to vary widely between the species from the 18 

genera studied, from trace amounts to 0.5%. In Leucojum vernum, Galanthus elwesii, 

G. nivalis, Ungernia victoris and Narcissus, galanthamine was found to be the principal 

component of the alkaloid fraction, making up from 30 to 50% of the alkaloid 

fraction and corresponding to 0.05–0.5% on a dry weight basis (Cherkasov, 1975; 

Gizba et al., 1982). 

Galanthamine is usually accompanied by lycorine in plants. Lycorine has been 

isolated from various species of the Amaryllidaceae family, including Amaryllis 

belladonna var. purpurea, Cooperanthes (Cooperia × Zaphyranthes) hortensis, Crinum 

defixum, C. laurentii, C. powelliti var. krelagei, C. yemense (latifolium), Eustephia yuyuensis,


Galanthus elwesii, G. nivalis var. gracilis, G. woronowii, Haemanthus katherinas, Hippeastrum 

hybridum ‘Salmon Joy’, H. rutilum, Hymenocallis amancaes, H. calathina, 

H. rotata (lacera), Ismene (Hymenocallis) hybridum ‘Sulphur Queen’, Leucojum aestivum, 

L. vernum, Lycoris albiflora, L. aurea, L. incarnata, L. radiata, Narcissus × gracillis, 

N. × incomparabilis, N. lobularis, N. ‘odorus rugulosus’, hybrids derived from N. jonquilla, 

N. poeticus, N. pseudonarcissus, N. tazetta and N. triandrus, Nerine flexuosa, 

N. undulata, Pancratium illyricum, Sternbergia fischeriana, Ungernia victoris, Vallota 

purpurea, Zephyranthes andersonii (andersoniana) and Z. rosea (Abdusamatov et al., 

1969; Boit, 1961; Cherkasov, 1976, 1977; Kadyrov et al., 1980; Medvedeva et al., 

1994; Popova, 1982; Ruan, 1988). In Pancratium trianthum and Hymenocallis littoralis 

from the Upper Volta and Ivory Coast, lycorine was the predominant alkaloid 

(Frederik, 1982). Interconversion of ungiminorine and lycorine was found in 

Ungernia severtsovii. Towards the end of the growing period, the level of lycorine in 

roots and bulbs increased, while that of ungiminorine decreased. At the time of 

leaf development this was reversed (Smirnova et al., 1965). 

SEARCHING FOR NOVEL SOURCES OF GALANTHAMINE 

Amaryllidaceae in the flora of the Russian Federation 

and adjoining regions 

Amaryllidaceae are relict species, believed to have been widespread in periods of 

tropical climate and retreating in glacial periods. Representatives of the Amaryllidaceae 

growing in temperate climates are thought to be the most advanced species. 

Some Amaryllidaceae occur in European Russia and the adjacent countries, and 

they are found in the Central Asia region (Artyushenko, 1970). Galanthus and 

Leucojum are found throughout Russia and the adjoining countries (Khokhryakov, 

1966), and are traditionally grown in gardens and greenhouses. 

The Amaryllidaceae family is represented in the flora of the Russian Federation 

and adjoining regions by seven genera: Leucojum L., Ixiolirion Herb., Narcissus L., 

Pancratium L., Galanthus L., Ungernia Bge. and Sternbergia Waldst. and Kit. (Artyushenko, 

1965; Kalashnikov, 1970; Komarov et al., 1935). 

Leucojum has ten species found from Ireland and North Africa to the Crimea 

and Caucasus, including two species in southern Ukraine, the Transcaucasian 

region and northern Caucasus. Ixiolirion has five species growing widely in Asia, 

while two of them are found in the Caucasus, Central Asia and eastern Siberia. 

Narcissus includes 25 to 30 species distributed in Europe and Central Asia. Pancratium 

contains 14 species, one of them growing in the region of the Black Sea coast. 

Galanthus species are found in southern and central Europe and Asia Minor. The 

majority of the species, 16 among a total of 25 to 27, grow widely in the Caucasus. 

Ungernia species (six populations) are found in central Asia (Artyushenko, 1970; 

Vvedensky, 1935, 1963; Korotkova and Khamidkhodzhaev, 1976). Sternbergia 

comprises three or four species distributed in the Black Sea region, the eastern 

Caucasus and Turkmenistan. 

Amaryllid species are perennial herbs, and their alkaloids occur mainly in the 

storage organs. In most Amaryllidaceae the above-ground parts of the radical 

leaves occur in files, the leaf bases forming the bulbs. Species of Amaryllidaceae


have from two to 15 leaves which are 7 to 80 cm long. Bulb diameters vary from 

1cm in Galanthus species to 12 cm in Ungernia. The inflorescences of the majority 

of Amaryllidaceae species are borne on leafless stems with between one (Galanthus 

sp.) and 25 florets (Ungernia). The florets occur in umbels or corymbs; the perianth 

is petalloid with six free or joined segments, there are six stamens in two whorls, 

and the fruit is a capsule (Artyushenko, 1970; Kalashnikov, 1970; Khamidkhodzhaev, 

1967, 1977). Unlike the other genera, Ixiolirion species have a tuberous storage 

organ derived from an enlarged stem base, and the leaves are borne on the aerial 

part of the stem. 

Collecting galanthamine-containing plants 

Because of the need to exploit Amaryllidaceae species which have a limited natural 

distribution – some species have been included in the ‘Red Book’ of the USSR and 

the Russian Federation (Takhtadjan, 1975, 1981; Borodin, 1978; Golovanov, 

1988) – a programme of collection and cultivation of suitable species was started 

by the All-Russian Institute of Medicinal and Aromatic Plants (VILAR). A comparative 

biological and chemical study of the Amaryllidaceae in the flora of Russia and 

the adjoining territories was carried out to find species suitable for commercial 

exploitation. The collection of plant material was started in 1975. Seeds of Amaryllidaceae 

plants were obtained from ‘delectus seminum’ (seed exchanges via air-mail), 

while seedling material of narcissus was obtained from commercial flower growers 

and from botanical gardens. Specimens from the flora of Russia and the adjoining 

territories were collected from their respective regions by expeditions organised 

by VILAR and its regional experimental agricultural stations. Some of the experimental 

results are given below. 

Galanthus woronowii containing up to 0.78% galanthamine was found in Adjaria 

in a canyon of the Chakrizkaly river (Patudin et al., 1978). It is distributed in the 

area of the Caucasus as far as the Turkish border. However, large-scale cropping 

was not practical, because the plant has such a low weight. Another galanthaminecontaining 

representative of the family is Leucojum aestivum which grows widely on 

the sea-coasts of the Caucasus, occupying a vast area in Colkhida and also in 

Ukraine and Moldavia (Transcarpathia). L. aestivum contains tenths or hundredths 

parts of one per cent of galanthamine, depending on where it is collected (Cherkasov, 

1975). Galanthus nivalis was collected in Ukraine in the Carpathian Mountains, 

the basin of the Dnieper River. Leucojum vernum and Narcissus angustifolius were 

collected in Ukraine. Ungernia victoris was reported in the region of Uzbekistan 

and Tajikistan (Korotkova and Adylov, 1960). 

Early in 1975, the collection of galanthamine-containing plants consisted of 

seven species and 245 populations and forms, including ten populations of 

Galanthus woronowii, two of G. nivalis, ten of Leucojum vernum, 48 of L. aestivum, four 

of Ungernia victoris and 163 of Narcissus (including 115 varieties; see below) 

(Cherkasov et al., 1984b; Kiselev et al., 1977). 

Species suitable for commercial exploitation 

During studies on the species mentioned above, the concentration of galanthamine 

was the main factor taken into consideration. Also considered was


information on biomass accumulation and growth cycles, and the influence of 

climatic and geographic parameters. High galanthamine levels were not always 

accompanied by other desirable characteristics. Over 1100 specimens of wild 

species and cultivars, both aerial and underground parts, were examined for 

galanthamine content. 

The best combination of characteristics (high galanthamine content and high 

biomass production) was found in Ungernia victoris, natural areas of which are 

presently exploited for the commercial production of galanthamine. However, 

attempts to cultivate U. victoris, a species suited for growing on hillsides, met with 

difficulties. U. victoris grows widely in the Gissar mountain range of Tajikistan. It 

had an alkaloid level of 0.05–0.60% in bulbs and 0.05–0.15% in leaves. However, 

the maximum accumulation of total alkaloids and galanthamine in the green parts 

of the plant was found in the phase of early growth in spring, when leaf length did 

not exceed 1–5 cm. Thus, the weight of green mass in wild plants was insufficient 

for commercial exploitation (Abduazimov and Yunusov, 1960; Abdusamatov, 

1972; Abdusamatov et al., 1963; Khamidkhodzhaev, 1967, 1977, 1982, 1984; 

Cherkasov et al., 1984b). 

Galanthus woronowii is characterised by a high content of galanthamine in the 

majority of wild populations and cultivars with 0.10–0.9% in bulbs and 0.05–0.70% 

in leaves. However, Galanthus species have no prospects for commercial culture, as 

the plant has a low mass, is easily damaged during mechanical cultivation, and 

does not adapt well to novel climatic conditions (Kovtun et al., 1978; Patudin et al., 

1978). 

Leucojum vernum grows widely in the Karpaty region and Ukraine, while over 

30 populations of L. aestivum are distributed in the Transcaucausian region. They 

contain from trace amounts up to 0.15–0.18% of galanthamine in bulbs and leaves. 

The leaves of L. aestivum populations (from Abkhazia) were found to be the most 

potentially useful, as a source for drug production, because in some populations 

there was 0.30% galanthamine in bulbs and 0.34% galanthamine in leaves 

(Cherkasov, 1975; Cherkasov et al., 1984b; Gizba et al., 1982). The biology of seed 

germination and seedling growth in L. aestivum has also been studied (Cherkasov, 

1980). The dynamics of galanthamine accumulation in L. aestivum was studied 

by Stefanov et al. (1974), and it was found that maximum concentrations of the 

alkaloid occurred in the phase of bud formation. The natural area of growth and 

supply of this species, used in Bulgaria for the production of galanthamine, has 

been determined by Stojanov and Savchev (1964). An experimental study on the 

introduction of Galanthus, Leucojum, Narcissus and Ungernia in the Moscow region 

has been carried out by Kiselev et al. (1977). 

Narcissus species and cultivars as prospective sources 

for galanthamine production 

Narcissus have been used for decoration since ancient times in Iran, Greece, Rome 

and Egypt, and has been described in mythological sources. Its garden forms have 

been systematically studied by the present authors for galanthamine content. 

There are about 60 species of Narcissus growing widely in Europe and in the 

Mediterranean region. In ‘The Flora of USSR’ (Komarov et al., 1935; Kuvaev and 

Khamidkhodzhaev, 1989) about 20 species were described, and ‘The Classified


List and International Register of Daffodil Names’ (RHS, 1975) contains over 

9000 names. Narcissus populations growing in the Caucasus are believed to be 

natural forms. Narcissus species possess a considerable vegetative mass, exceeding 

that of Galanthus woronowii. 

Galanthamine content was determined for 118 varieties of Narcissus. In 33 

garden cultivars, the level of galanthamine exceeded 0.1% of the dry weight of 

the plant. In wild populations, the concentration was almost zero. 

A collection of Narcissus species and cultivars has been maintained for a 

number of years in the introduction plots of VILAR. Analysis of the alkaloid 

content of bulbs of 26 narcissus revealed that galanthamine was present in all 

samples (Cherkasov et al., 1985). Of 19 Narcissus varieties, 15 contained foliar 

galanthamine. The cultivar ‘Favourite’ contained 0.15% on a dry weight basis, 

whereas in other cultivars the galanthamine content ranged from trace levels to 

0.09%, fluctuating from year to year (Cherkasov et al., 1986). At flowering time, 

leaves and bulbs of narcissus cultivars growing in the field contained 0.02–0.10 

and 0.05–0.10% galanthamine (dry weight basis), respectively. The alkaloid 

contents in five greenhouse cultivars were correspondingly 0.10–0.20 and 0.05– 

0.20% (dry weight). Thus, after harvesting the flowers, the leaves may be used 

for galanthamine production (Maisuradze et al., 1985). Of 81 cultivars of 

narcissus, 72 contained galanthamine in both leaves and bulbs. Galanthamine 

was reported for the first time in 79 narcissus cultivars (Cherkasov et al., 1988, 

1993). The galanthamine concentration in the crude drug was dependent on the 

plant variety, period of harvest, weather conditions, etc. The highest level of 

galanthamine was found in leaves of eight populations, up to 0.2–0.5%. The 

highest galanthamine content was found at the phase of bud formation when the 

growth rate was the highest. 

Narcissus ‘Fortune’ was proposed as a prospective subject for cultivation for 

galanthamine extraction, as it was characterised by a consistency of chemical 

composition. This cultivar is widely used in commercial floriculture (Cherkasov 

et al., 1989; Maisuradze et al., 1985; Zaitseva and Novikova, 1977). The alkaloid 

composition of leaves and bulbs, and the dynamics of galanthamine accumulation 

during the growing period, were studied for ‘Fortune’. Galanthamine, 

gemanthamine and a novel alkaloid named fortucine (Figure 8.1) were identified 

among the five alkaloids isolated. The structure of fortucine was elucidated by 

nuclear magnetic resonance and mass spectrometry, the latter distinguishing 

its structure from the earlier known lycorine-type alkaloids by the atypical 

cissoid fusion of the B/C rings (Gorbunova et al., 1984; Tokhtabaeva et al., 

1987). The seed germination of Narcissus species has been studied by Cherkasov 

(1982). 

Drying regimes for narcissus leaves have been studied, and the optimal parameters 

for galanthamine production were determined (80 °C, 0.77 kg/m 2 /h) 

(Voroshilov et al., 1987, 1989). 

Cell cultures of the above plants were also studied. They were proposed for 

clonal and accelerated reproduction of rare species (Popov and Cherkasov, 1983, 

1984). It has been shown that ‘shoot-clump’ cultures of Narcissus confusus in liquid 

medium were capable of producing galanthamine (Bastida et al., 1996). Treatment 

with sucrose increased the growth of the cultures and affected galanthamine 

biosynthesis (Selles et al., 1997b).


ANALYTICAL METHODS OF GALANTHAMINE DETERMINATION 

IN AMARYLLIDACEAE EXTRACTS 

Earlier methods for galanthamine determination in plant material used paper 

chromatography (Vulkova and Kolusheva, 1964) and spectrophotometry (Asoeva 

and Bergeichik, 1967; Vulkova, 1959; Volodina et al., 1970; Kalashnikov et al., 

1980; Kuznetsov et al., 1969). However, a direct spectrophotometric determination 

of galanthamine is limited in its usefulness, as the compound has a low specific 

absorption index and occurs in the plant at low concentrations. Kolusheva and 

Vulkova (1966) have used spectrophotometric methods for the estimation of 

galanthamine (Nivaline) in ampoule solutions. 

A polarographic method of galanthamine determination, proposed for measuring 

the compound in leaves of Ungernia victoris, was found experimentally to have low 

accuracy (Volodina et al., 1970). 

Methods of quantitative determination based on chromatographic or chromatoextractive 

separation and photo-colorimetric or photometric estimation of 

galanthamine in leaves of Ungernia victoris have been reported (USSR Pharmacopoeia, 

11th edition), which were later modified and adapted for the quantitative 

analysis of Narcissus ‘Fortune’ (Tokhtabaeva, 1987), N. poeticus (Popova and 

Karpenko, 1989), other garden narcissus varieties and other Amaryllids such as 

Galanthus and Leucojum. In the method of Tokhtabaeva (1987), plant material was 

extracted in an acetone – water – 25% ammonia mixture. After transfer to chloroform 

the extract was separated by thin-layer chromatography on silica gel using 

chloroform – ethyl acetate – 25% ammonia as solvent. The detected zone was 

treated with tropaeolin 000 Nr 2 (Orange II) and the complex with the alkaloid 

was extracted with chloroform. The optical density of the solutions was measured 

photocolorimetrically. The method was used in the analytical control of galanthamine 

hydrobromide production. A semi-quantitative express method was also 

developed (Tokhtabaeva, 1987). 

Recently, a high-performance liquid chromatography method has been published 

for the separation and quantitative determination of galanthamine and 

other Amaryllidaceae alkaloids in plant extracts and tissue cultures of Narcissus 

confusus (Selles et al., 1997a). Capillary gas chromatography was used for the determination 

of the distribution of galanthamine and other alkaloids in Narcissus ‘Ice 

Follies’ (Moraes-Cerdeira et al., 1997a,b). 

A method of galanthamine extraction from leaves of Narcissus ‘Fortune’ with 

aqueous organic solvents has been elaborated with the application of mathematical 

models for the optimisation of the process (Rusakova et al., 1986). An efficient 

large-scale technological method of galanthamine hydrobromide production has 

been worked out utilising available solvents and materials (Tolstykh et al., 1991, 

1992). Shakirov et al. (1969) have developed an ion exchange method of galanthamine 

hydrobromide production at a commercial scale from Ungernia victoris 

Vved. in 80% yield with the application of Ku-1 resin. 

CONCLUSIONS 

The alkaloids galanthamine and lycorine are valuable constituents of Amaryllidaceae 

species widely used in medicine. Recent progress in treatment of


Alzheimer’s disease has attracted scientists for new investigations in this field. 

Comparative studies of various genera of this family have been carried out, and 

prospective species were found for commercial exploitation, among them Leucojum, 

Ungernia and Narcissus. The technological procedures for their production have 

been elaborated. 

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Narcissus cv. Ice Follies. Planta Medica, 63 (1), 92–93. 

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of Flowering Decorative Plants, Moscow, pp. 14–19 (in Russian).

9 Studies on galanthamine extraction 

from Narcissus and other 

Amaryllidaceae 

Mirko Kreh 

INTRODUCTION 

When in 1947 Proskurnina and Yakovleva isolated a novel alkaloid, which they 

named galanthamine, from Galanthus woronowii (Caucasian snowdrop), they could 

not have foreseen the relevance of their findings for the pharmaceutical industry. 

Success in the treatment of poliomyelitis with galanthamine was soon reported, 

and the alkaloid became a routine part of the Soviet stock of medicines. As early as 

1960, it was reported that galanthamine was an inhibitor of acetylcholinesterase, 

similar to the natural product physostigmine and the synthetic product neostigmine 

(Schulz, 1960). 

Medicinal and pharmacological tests led to a huge number of potential uses for 

galanthamine. Among the variety of indications cited in the literature (ABDA, 

1993) are described analgesic effects comparable with morphine (Cozanitis and 

Rosenberg, 1983), compensation of the effect of opiates on respiration (Paskov 

et al., 1963), effects on alcohol abuse (Opitz, 1991) and effects in treating Alzheimer’s 

disease (Han et al., 1991). 

Until the 1960s, galanthamine was isolated from natural supplies of snowdrop, 

Galanthus nivalis (Chimiko-Pharmazevtitschen Zavod, 1959), but commercial 

production from Ungernia species was also described (Cherkasov et al., 1986). 

Nowadays, galanthamine is also isolated from Leucojum aestivum (summer snowflake) 

(Gorinova et al., 1993), although a number of promising chemical syntheses have 

been reported (Czollner et al., 1998). 

The promising pharmacological properties of galanthamine, and the shrinking 

natural sources of the species mentioned, made it of interest to find new sources of 

galanthamine within the Amaryllidaceae family. The research reported here is 

based on a doctoral thesis presented to Marburg University (Kreh, 1995). 

SELECTION AND SCREENING OF PLANT MATERIAL 

The search for galanthamine was restricted to the Amaryllidaceae, because this 

family has a very specific alkaloid metabolism. In the literature data concerning 

galanthamine and the alkaloid content of species of Amaryllidaceae are found in 

many publications (e.g., Fuganti, 1975; Cherkasov, 1977; Cherkasov et al., 1984, 

1986, 1988, 1989; Tanahashi et al., 1990). The concentrations reported for assay 

often differ for the same species. From the literature it was not possible to decide

Galanthamine extraction 257 

whether these differences in galanthamine content resulted from extracting plants 

at different stages of development or from using plants cultivated in different 

regions, or were simply due to the low purity of the isolated alkaloid. 

The screening described below was restricted to bulb tissue, because bulbs are 

the usual commodity traded and, unlike the aerial parts of the plants, are available 

in large quantities. The genus Narcissus was of particular interest, since it is indigenous 

to Europe and material can easily be obtained in quantity from large 

daffodil cultivars. Narcissus have been described as sources for natural galanthamine 

several times (Cherkasov, 1977; Cherkasov et al., 1984, 1986, 1988, 1989). 

Looking specifically for plants which would be available in large quantities, and 

therefore at a low price, the screening was restricted mainly to the 20 daffodil 

cultivars most commonly grown in the Netherlands (Erhardt, 1993). 

The plant material was obtained from cultivars grown around Hillegom and on 

Texel Island, the Netherlands. The identity of the plants was checked during the 

flowering period by studying their anatomical and microscopic characteristics. 

The daffodil cultivars used were identified according to ‘The International Daffodil 

Checklist’ of the Royal Horticultural Society (Kington, 1989), and, as far as possible, 

from reference plants and photographs. 

High-performance liquid chromatography (HPLC) (Kreh, 1995) and gas 

chromatography (Kreh, 1995; Kreh et al., 1995) were found to be suitable 

analytical methods for the analysis of alkaloids from Amaryllidaceae. For the 

quantitative galanthamine screening described below, the following HPLC 

method was used: 

Stationary phase: LiChrospher® 60 RP-select B, 5 µm, 250 × 4 mm column 

(Merck) 

Eluent A: 2.0 g sodium dodecylsulphate, 2000 ml water, 400 ml acetonitrile and 

290 ml 0.05 M phosphoric acid 

Eluent B: acetonitrile 

Gradient: 0–25 min 25:75 A:B, 30–45 min linear to 100:0, flow 1ml/min 

For a detailed description of sample preparation, analytical equipment and quantitative 

evaluation, see Kreh (1995). 

The results of the HPLC screening are given in Table 9.1. From the plants 

tested, several narcissus cultivars were found to be rich in galanthamine. 

However, in the case of the Galanthus and Leucojum bulbs tested, only low concentrations 

of galanthamine were found. Differences between these values and 

those reported in the literature might result from the occurrence of different 

chemical races (Schulz, 1960) or from the effects of growing on different soils 

(see below and Gorinova et al., 1993). Such differences were also found in 

screening Narcissus ‘Fortune’ from Texel Island and from Hillegom, which have 

different soil types. 

Narcissus ‘Carlton’, ‘Gigantic Star’, ‘Ice Follies’ and ‘Fortune’ were found to be 

rich in alkaloids and galanthamine. All of these belong to the group of ‘largecupped’ 

daffodils (Division 2; Kington, 1989), which might be interesting for a 

further search for narcissus rich in galanthamine. 

For all the subsequent studies Narcissus ‘Carlton’ (Figure 9.1) was used, since it 

fulfils nearly all the requirements for galanthamine production on a large scale. 

It has been cultivated for many years in large amounts in the Netherlands and

258 M. Kreh 

Table 9.1 Results of galanthamine screening: percentage content of total alkaloids and 

galanthamine expressed on a fresh (FW) or dry (DW) weight basis 

Species or cultivar % Alkaloids % Galanthamine 

FW DW FW DW 

Chlidanthus fragrans 0.371 0.531 0.0017 0.0054 

Crinum × powellii 0.200 0.231 0 0 

Elisena longipetala 0.090 0.162 0 0 

Galanthus elwesii 0.113 0.290 0.0084 0.0213 

Galanthus nivalis 0.071 0.124 0.0013 0.0031 

Ixolirion pallasii 0.003 0.007 0 0 

Leucojum aestivum 0.033 0.236 0.0009 0.0065 

Leucojum vernum 0.034 0.107 0.0001 0.0001 

Narcissus ‘Barrett Browning’ 0.127 0.263 0.0047 0.0096 

Narcissus ‘Brougshane’ 0.172 0.503 0.0004 0.0012 

Narcissus ‘Carlton’ 0.216 0.555 0.0725 0.1880 

Narcissus ‘Dick Wilden’ 0.099 0.230 0.0008 0.0019 

Narcissus ‘Dutch Master’ 0.176 0.524 0.0001 0.0001 

Narcissus ‘February Gold’ 0.105 0.660 0.0118 0.0780 

Narcissus ‘Flower Drift’ 0.109 0.254 0.0163 0.0381 

Narcissus ‘Flower Record’ 0.177 0.927 0.0150 0.0800 

Narcissus ‘Fortune’ (from Hillegom) 0.140 0.310 0.0224 0.0488 

Narcissus ‘Fortune’ (from Texel) 0.174 0.583 0.0208 0.0695 

Narcissus ‘Gigantic Star’ 0.157 0.650 0.0290 0.1201 

Narcissus ‘Golden Harvest’ 0.161 0.407 0 0 

Narcissus ‘Ice Follies’ 0.090 0.264 0.0250 0.0740 

Narcissus ‘Minnow’ 0.028 0.079 0.0006 0.0018 

Narcissus ‘Salome’ 0.073 0.139 0.0020 0.0038 

Narcissus ‘Tête-à-Tête’ 0.066 0.168 0 0 

Narcissus ‘Unsurpassable’ 0.225 0.591 0.0004 0.0010 

Narcissus ‘Van Sion’ 0.089 0.206 0.0037 0.0086 

Narcissus ‘Verger’ 0.055 0.161 0 0 

Narcissus ‘Yellow Sun’ 0.138 0.480 0.0035 0.0123 

Nerine bowdenii 0.220 0.241 0 0 

Sprekelia formosissima 0.011 0.021 0 0 

Zephyranthes robusta 0.070 0.090 0.0033 0.0150 

the UK with 354 ha in 1988/1989, it was the most cultivated narcissus in the 

Netherlands, and therefore easy to obtain and cheap (Erhardt, 1993). Narcissus 

‘Carlton’ is not especially susceptible to diseases and climate, special winter protection 

is not necessary, and it has a high rate of multiplication (Jefferson-Brown, 

1991). 

Since nearly all Amaryllidaceae alkaloids previously tested had interesting 

pharmacological and biological properties, it was the intention to isolate as many 

different alkaloids as possible from Narcissus ‘Carlton’. Using preparative HPLC, 

twenty-nine alkaloids were isolated from the bulbs, in addition to galanthamine. 

The structures of the then unknown natural products 1-O-acetyl-10-norpluviine, 

10-norpluviine, O-methyloduline, N-demethylmasonine, 1,10-diacetyl-10-norpluviine 

and O-acetylgalanthamine were elucidated by modern spectroscopic techniques 

as well as by conventional analytical methods (Kreh, 1995).

Figure 9.1 Narcissus ‘Carlton’. 


CONTENT AND DISTRIBUTION OF GALANTHAMINE IN NARCISSUS 

‘CARLTON’ THROUGH THE YEAR 

It is obvious from the growth pattern of narcissus that the amount of plant material 

available for galanthamine extraction varies greatly through the year, so it was 

presumed that galanthamine content within the plant might also vary during the 

year. It was already known that bulbs of narcissus harvested in March had significantly 

lower anti-tumour activity than bulbs from the same variety harvested 

between July and December (Fitzgerald et al., 1958). To determine if there is an 

optimum time for the extraction of ‘Carlton’ bulbs for galanthamine, bulbs were 

planted in the old botanical garden of Marburg University. Over a twelve month 

period, samples were taken up to twice per week and tested for alkaloid and galanthamine 

content. The key developmental dates for these plants were: 

Bulbs planted: 27 September 

First leaves emerged: 24 February 

Start of flowering: 30 March 

Crop in full flower: 19 April 

Capsule dehiscence: 20 June 

Leaves died down: 18 August 

Using a large number of samples and with HPLC as a highly specific analytical 

technique, significant results were obtained. Besides the quantitative determination 

of galanthamine content, analytical HPLC gave very informative findings in

260 M. Kreh 

relation to the qualitative and quantitative composition of the other alkaloids 

extracted. 

Galanthamine content of the bulb 

Figure 9.2 shows the content of galanthamine in the bulb of Narcissus ‘Carlton’. 

Galanthamine content decreased slightly over the first few samples (in November). 

Three months after planting, galanthamine content increased over about four 

Figure 9.2 Galanthamine content of bulbs of Narcissus ‘Carlton’ over one year. Upper 

curve calculated on a dry weight basis; lower curve calculated on a fresh 

weight basis.


weeks by more than 400%, calculated on a fresh weight basis, peaking in April. 

The galanthamine content of the bulb decreased from the middle of April until 

the beginning of June, and increased again in parallel with leaf senescence. Therefore 

the optimum time for harvesting and extracting bulbs would be between the 

end of July and the beginning of August. Bulbs intended for galanthamine extraction 

should not be lifted earlier than this. 

Although there are changes in galanthamine content of the bulbs through the 

growth cycle, it is not known whether some of the differences observed during the 

first weeks of the study were due to the change in location of the bulbs, which were 

obtained from Sint Maarten, the Netherlands (with a sandy soil) and grown at 

Marburg (in a loamy soil). 

Galanthamine content of the aerial parts 

During this experiment, the aerial parts of the plants were investigated as well as 

the bulb. Figure 9.3 shows the galanthamine content in the whole aerial parts of 

Narcissus ‘Carlton’. With the growth of the leaves in spring, galanthamine content 

increased dramatically until just before the start of flowering at the end of March, 

and decreased slowly until the end of the flowering period. It then remained more 

or less constant for some weeks, reaching the lowest level when the leaves had 

completely died down. 

The galanthamine content of the aerial parts was significantly lower than that of 

the bulb, calculated on a fresh weight basis, but it was still in a range which would 

make it useful for technical extraction. However, the removal of the leaves of 

the growing crop would not be practical because of the likelihood of subsequent 

rotting of the bulbs. 

Location of galanthamine in the plant 

Several plants of Narcissus ‘Carlton’ were separated during the flowering period 

into bulb, roots, leaves, scape (stem) and flower. These parts made up the following 

percentages of the whole plant: bulb, 39.8%; roots, 3.5%; leaves, 29.2%; scape, 

21.9% and flower, 3.8%. After the natural opening of the capsules, the seeds of 

several plants were also collected. All plant parts were tested for their content of 

alkaloids and galanthamine. 

Figure 9.4 shows the results of the galanthamine assay. The results demonstrated 

that the bulb is clearly the most interesting part of narcissus for galanthamine 

extraction, although all parts of the plant contain alkaloids and, in 

particular, galanthamine. It can be calculated from the weights of the different 

plant parts that bulbs planted in September (average weight, 78 g) contained an 

absolute amount of galanthamine which is higher than in the whole plant during 

its flowering period (average weight, 140 g). In summer, the absolute amount of 

galanthamine increased again, when the new bulb units (‘daughter-bulbs’) were 

growing. 

Analytical HPLC allows an overview about the composition of the alkaloids in 

the individual plant parts (Figure 9.5). Galanthamine is the main component only 

in the bulb. In the scape, leaves and seed haemanthamine prevails, while in the 

flower and root both galanthamine and haemanthamine are prominent.

262 M. Kreh 

Figure 9.3 Galanthamine content of aerial parts of Narcissus ‘Carlton’ over one year. 

Upper curve calculated on a dry weight basis; lower curve calculated on a 

fresh weight basis. 

EFFECT OF FERTILISERS ON THE GALANTHAMINE 

CONTENT OF NARCISSUS ‘CARLTON’ 

The effect of different fertilisers on narcissus growth is well known among bulb 

growers. Potassium helps to develop strong bulbs and good flowers, phosphorus 

leads to good root development and strong plants, and nitrogen supports the 

growth of the leaves and the development of the bulb after leaf senescence. Too

% Galanthamine 

0.040 

0.035 

0.030 

0.025 

0.020 

0.015 

0.010 

0.005 

0.000 


Flower Scape Leaf Root Bulb Seed 

Figure 9.4 Galanthamine content of the different plant organs of Narcissus ‘Carlton’ on 

a fresh weight basis. Determined for flowering plants on 27 April, except 

for seed which was analysed after opening of the capsules (27 June). 

much nitrogen, however, leads to ‘soft’ bulbs, which are sensitive to plant diseases 

and often rot (Erhardt, 1993). It was possible that the different screening results 

obtained for the galanthamine content of Narcissus ‘Fortune’ from Hillegom and 

Texel Island, the Netherlands (see Table 9.1) were a result of using different fertilisers 

or different soil types. Therefore, the effect of different commercial fertilisers 

on the galanthamine content of Narcissus ‘Carlton’ was examined. 

Plots of narcissus were laid out in the old botanical garden of Marburg University. 

Analysis of the soil gave results typical of sandy, loamy soil (pH 6.3 and nutrient 

levels (mg/100 g soil) of 41 P 2 O 5 , 16 K 2 O, 11 Mg and 0.15 soluble N). One of the 

plots was treated with 500 g N fertiliser per m 2 (‘Kalk-Ammon-Salpeter’, 27% N, of 

which 13.5% nitrate-nitrogen and 13.5% ammonium-nitrogen), and another with 

500 g K-Mg fertiliser per m 2 (30% K 2 O, 10% MgO). Untreated control plots were 

also grown. These amounts of fertiliser were found to be excessive, and several 

plants did not flower during the first year and the bulbs rotted. However, nearly 

all of the plants developed normally in the second year. Figure 9.6 shows the 

results of the galanthamine determination in the second year. The use of either 

nitrogen or potassium/magnesium fertiliser increased the galanthamine content of 

bulbs and leaves significantly over the control. An increase in galanthamine content 

by 70% was achieved in the case of the nitrogen fertiliser, and of 113% after 

using potassium/magnesium fertiliser. The results clearly demonstrated the effect 

of fertilisers on galanthamine content. Not only was the total amount of alkaloids 

markedly increased by fertiliser treatment, but also the percentage of galanthamine 

in the alkaloid fraction.

264 M. Kreh 

2.0 

flower 

2 

1 

3 

4 

0 10 20 30 40 

1.3 

leaf 

1 

2 

3 

4 

0 10 20 30 40 

3.2 

root 

1 

2 

3 

4 

0 10 20 30 40 

5 

5 

5 

6 

6 

6 

scape 

ASPECTS OF A POTENTIAL TECHNICAL EXTRACTION 

OF GALANTHAMINE 

Narcissus ‘Carlton’ was identified as a potential source for technical galanthamine 

extraction. The well-known procedures for the production of alkaloids such as 

cocaine, morphine, atropine, chinine, etc., all include, as one step, a typical 

acid/base extraction (Schwyzer, 1931), as do the described procedures for the 

extraction of galanthamine (Chimiko-Pharmazevtitschen Zavod, 1963; Proskurnina- 

Shapiro and Yakovleva, 1966a,b; Paskov and Ivanova, 1967). The desired substance 

1.4 

1 

2 

3 

4 

0 10 20 30 40 

2.0 

bulb 

2 

1 

3 4 

0 10 20 30 40 

1.4 

seed 

1 

2 

3 

4 

0 10 20 30 40 

Figure 9.5 HPLC chromatograms of the alkaloids obtained from different parts of flowering 

Narcissus ‘Carlton’. 1 = N-demethylgalanthamine; 2 = galanthamine; 3 = lycorenine; 

4=O-methyloduline; 5 = haemanthamine; 6 = homolycorine. 

5 

5 

5 

6 

6 

6



0.018 

0.016 

0.014 

0.012 

0.010 

0.008 

0.006 

0.004 

0.002 

0.000 

0.050 

0.045 

0.040 

0.035 

0.030 

0.025 

0.020 

0.015 

0.010 

0.005 

0.000 


No fertiliser NH4 / NO3 Fertiliser K / Mg Fertiliser No fertiliser NH4 / NO3 Fertiliser K / Mg Fertiliser Figure 9.6 Galanthamine content in leaf (above) and bulb (below) of Narcissus ‘Carlton’ 

following the application of different fertilisers. All values calculated on a 

fresh weight basis. 

is crystallised or precipitated from the raw alkaloid fraction with a suitable solvent 

or reagent. 

To find a profitable extraction method for galanthamine, the following factors 

were taken into consideration: 

• Isolation using a simple procedure without expensive steps such as chromatography. 

• The isolation method has to guarantee a maximum yield of the desired substance.

266 M. Kreh 

• High purity of the resulting product. 

• No drying of the plant material in the process, since it could be shown that 

drying the bulbs decreases galanthamine content. 

• No use of expensive and toxic solvents during the entire procedure. 

Optimum solvent for extraction 

In the acidic plant sap galanthamine occurs in the form of its salts. A solvent was 

required that dissolves these salts or, after alkalisation of the plant material, the 

free galanthamine base. Tests were carried out using acetone, ethanol, methanol, 

isopropanol, chloroform, dichloromethane, diethylether, ethylacetate, n-heptane, 

n-hexane and n-pentane as the primary solvent for extraction and, following acid/ 

base work-up, for obtaining the raw alkaloids (before extraction with the nonwater-miscible 

solvents the material was made alkaline using sodium carbonate). 

The alkaloids were analysed by HPLC (see examples in Figure 9.7). 

The polar, water-miscible solvents tested led to a quantitative isolation of galanthamine 

even without alkalisation of the plant material. These solvents are easily 

miscible with the crudely crushed plant material. The main disadvantages were 

that the compounds extracted in parallel with the alkaloids caused persistent 

emulsions during the following work-up, and that the procedure showed no preference 

for galanthamine within the alkaloid fraction. 

0 1 2 3 4 5 

methanol 1 

0 5 10 15 20 

0 1 2 3 4 5 

0 1 2 3 4 5 

n-heptane 1 

0 5 10 15 20 

Figure 9.7 HPLC chromatograms of the alkaloid fractions obtained by using methanol 

(left) and n-heptane (right) for primary extraction of bulbs; peak 

1 = galanthamine. 

0 1 2 3 4 5


The apolar, non-water-miscible solvents tested also led, after alkalisation of 

the plant material, to a quantitative extraction of galanthamine. However, 

plant material and solvent had to be mixed vigorously, and thorough homogenisation 

of the bulbs greatly facilitated the process; without this, the sticky, wet 

plant material forms lumps into which the solvent cannot penetrate. The main 

advantage of some of the apolar solvents is a selectivity for galanthamine 

within the alkaloid fraction. For example, using chloroform for primary 

extraction, alkaloids were completely extracted. Using n-heptane, n-hexane or 

n-pentane for extraction, an alkaloid fraction was obtained in which galanthamine 

and haemanthamine were significantly enriched. For the following tests 

n-heptane was chosen, since it is less toxic and less inflammable than n-hexane or 

n-pentane. 

The solubility of the galanthamine base is remarkable: it can be extracted with 

polar solvents such as methanol as well as with apolar solvents such as n-heptane. 

A possible explanation for this is the formation of intramolecular hydrogen bonds 

between the hydroxyl proton and the oxygen atom of the ether group, which 

makes the molecule less hydrophilic for the solvent in apolar solvents (Carroll, 

1990). In polar solvents intermolecular hydrogen bonds can be formed which 

explain its solubility in this group of solvents. 

Optimum pH for extraction 

Since alkaloids may be very different in their basicity, the optimum pH value for 

the following liquid-liquid separations was studied. The crude alkaloid fraction 

was obtained by extraction with chloroform after alkalisation of the crushed 

plant material with sodium carbonate solution. The organic phase was extracted 

with diluted sulphuric acid and the resulting aqueous phase was adjusted to pH 

5.0 with diluted ammonia. The aqueous phase was extracted five times with 

diethyl ether. The pH of the aqueous solution was adjusted to pH 6.0 and the 

extraction was repeated five times using diethyl ether. The procedure was 

repeated in steps of one pH unit until pH 12.0 was reached. The resultant ether 

fractions (mainly alkaloids) were analysed by HPLC. It was shown that, at pH 5, 

galanthamine was not found in relevant amounts in the diethyl ether fraction. 

However many other substances, including several alkaloids, could be separated 

at this pH value. At pH values >6.0, galanthamine was found in larger amounts 

in the organic phase. At pH values >9.0, galanthamine was extracted quantitatively 

from the aqueous phase, in contrast to several very basic alkaloids which 

still remained in the aqueous phase at pH 9.0. It was concluded that, for a technical 

extraction of galanthamine from Narcissus ‘Carlton’, other alkaloids should 

be removed at pH 5.0 by extraction with an organic solvent, before a quantitative 

extraction of galanthamine from the aqueous phase at pH 9.0 with the 

organic solvent, is carried out. 

Selective crystallisation of galanthamine from the alkaloid mixture 

A series of tests with many commercially available solvents was conducted to find 

a suitable solvent for the crystallisation of pure galanthamine from the alkaloid

268 M. Kreh 

fraction of Narcissus ‘Carlton’. The best results were achieved using toluene and 

isopropanol. By crystallisation from isopropanol, galanthamine was obtained at 

a very high purity. 

Precipitation as galanthamine hydrobromide 

After precipitation of galanthamine from the alkaloid fraction using isopropanol, a 

significant amount of galanthamine remained in solution. To isolate the remaining 

galanthamine many experiments were carried out to precipitate galanthamine as 

various salts. However, none of the experiments proved better than the precipitation 

of galanthamine hydrobromide from a solution in acetone. This method is 

described in all patents concerning the isolation of galanthamine from plant 

material (Chimiko-Pharmazevtitschen Zavod, 1963; Proskurnina-Shapiro and 

Yakovleva, 1966a,b; Paskov and Ivanova, 1967). 

Formation of potential artefacts during the extraction process 

Potential consequences of the use of dichloromethane 

During efforts to find the best solvent for extraction of the plant material, it was 

found that the use of dichloromethane should be completely avoided. Using 

dichloromethane led to the formation of the quaternary ammonium salt (–)-N- 

(chloromethyl)-galanthaminium chloride (2) in considerable quantities as an artefact 

(Kreh et al., 1994). This compound was derived from galanthamine (1) by N-chloromethylation 

with the solvent dichloromethane: 

O 

MeO 

HO H 

N 

Me 

CH CI 

2 2 

HO H 

Under the same conditions using chloroform no reaction product was obtained. 

Potential consequences of an excessive use of acid 

During the extraction procedure it was intended to use sulphuric acid. To test for 

the potential degradation of galanthamine in this medium a sample of galanthamine 

was treated for 48 hours with 2% sulphuric acid at room temperature. In 

the HPLC chromatogram of the resulting mixture only extremely small amounts 

of degradation products were observed. However, it was of interest to know which 

potential degradation products would be formed under extreme conditions. 

O 

MeO 

(1) (2) 

- 

Cl 

+ 

N 

Me 

CH CI 

2


To obtain greater amounts of degradation products the experiment was repeated 

with 10% sulphuric acid at 70 °C. The components of the resulting solution were 

separated by preparative HPLC. 

It was found that the excessive use of sulphuric acid led to an epimerisation of 

galanthamine (1) at the C3-atom and the formation of epigalanthamine (4). This 

should be taken into consideration when literature reporting the isolation of 

epigalanthamine from plant material is found. Another compound isolated from 

the reaction mixture was 3,4-anhydro-galanthamine (3). The compound was not 

stable over a three week period. The formation of a second aromatic unit derived 

from galanthamine was found in the nuclear magnetic resonance (NMR) spectrum 

of a third degradation product which was isolated. This can occur only if a 

rearrangement in the galanthamine molecule takes place. A similar reaction is 

known after treatment of morphine with acid (apomorphine rearrangement). 

However, the methyl ether bond of galanthamine was not broken by sulphuric 

acid. The resulting molecule was identified as 6-O-methyl-apogalanthamine (5). 

The scheme on the following page shows the probable mechanism of the apogalanthamine 

rearrangement which leads to the formation of 6-O-methyl-apogalanthamine. 

CONCLUSIONS 

Thirty commercially available species and cultivars from the Amaryllidaceae family 

were tested for their galanthamine content. Several large-cupped daffodil cultivars 

were found to be rich in galanthamine, in particular Narcissus ‘Carlton’, which is 

cultivated on a large scale in the Netherlands and the UK. This cultivar proved to 

be a suitable subject for further studies on a potential galanthamine extraction. In 

this cultivar galanthamine was found to be present in all parts of the plant. The 

highest content was found in the bulb, which is convenient for technical extraction. 

Galanthamine concentration in the plant fluctuated strongly during the year. 

The best time for harvesting the bulbs would be a short time after the usual lifting 

period in the Netherlands. The content of galanthamine in bulbs could be 

increased by applying fertilisers. 

For isolation of galanthamine on a technical scale the following procedure is recommended. 

The bulbs are thoroughly homogenised with dry sodium carbonate 

and extracted with petroleum spirit (which is cheaper than n-pentane, n-hexane 

or n-heptane). The organic phase is concentrated by evaporation and extracted 

with diluted sulphuric acid. The acidic phase is adjusted to pH 5.0 with ammonia 

solution immediately after extraction, and separated from accompanying substances 

by extraction with diethyl ether. The aqueous phase is adjusted to pH 9.0 

and extracted again with diethyl ether. The ether extract is dried and dissolved in 

warm isopropanol. Crude galanthamine base crystallises during cooling, using a 

seed crystal if necessary. Pure galanthamine is obtained by re-crystallisation of the 

crude product from isopropanol. Galanthamine remaining in the mother liquids 

of the crystallisations is obtained after evaporating the remaining solution in 

isopropanol, dissolving the residue in acetone and precipitation with ethanolic 

hydrogen bromide solution. The procedure is simple, relatively fast and consists of

H 

O 

MeO 

O 

MeO 

H 

H 

HO H 

(1) 

(3) 

+ 2 H + 

N Me 

N Me 

N Me 

H O 

+ 

+ 

MeO 

HO 

MeO 

H 

+ 2 H + 

- 2 H 2 O 

- 2 H + 

O 

MeO 

H 

O 

MeO 

HO 

MeO 

(3a) (3b) 

(5) 

N 

Me 

- 2 H + 

HO 

MeO 

H 

+ 

+ H 2 O 

- 2 H + 

H OH 

(4) 

+ 

+ 

(3c) 

H 

H 

N Me + 

H 

N Me 

N Me + 

H 

rearrangement 

N Me + 

H 

(1a) 

+ 

(1) 

[ (4) : (1) about 7 : 1 ]


only a few steps. Expensive purification procedures such as chromatography could 

be completely avoided, and the use of problematic reagents is very limited. This 

method has been tested with 96 kg of bulbs of Narcissus ‘Carlton’, leading to a 

further patent (Hille et al., 1996). Galanthamine can be obtained in even higher 

purity than the drug currently available on the market. 


The author would like to acknowledge the help and support of Prof. Dr. R. Matusch, 

who served as mentor for this project. I would also like to thank F. van Elsäcker 

for his help in harvesting and extracting the plant material, and Lohmann Therapie- 

Systeme GmbH for financial support of the project. 

REFERENCES 

ABDA (1993) Central Database of the German Pharmacists Association, ABDATA-Pharma- 

Service, Frankfurt, Eschborn. 

Carroll, P., Furst, G.T., Han, S.Y. and Joullié, M. (1990) Spectroscopic studies on galanthamine 

and galanthamine methiiodide. Bulletin de la Société Chemique de France, 127, 

769–780. 

Cherkasov, O.A. (1977) Plant sources of galanthamine. Khimiko-Farmatsevticheskii Zhurnal, 11 

(6), 84–89 (in Russian). 

Cherkasov, O.A., Gorbunova, G.M., Margvelashvili, N.N., Maisuradze, N.I. and Tolkachev, 

O.N. (1986) The content of galanthamine in leaves of Narcissus hybridus hort. Rastitel’nye 

Resursy, 22 (3), 370–372 (in Russian). 

Cherkasov, O.A., Maisuradze, N.I., Glyzina, G.S. and Gayevskii, A.V. (1989) Narcissus as a 

source of raw materials for producing galanthamine. Khimiko-Farmatsevticheskii Zhurnal, 

23 (5), 621–623 (in Russian). 

Cherkasov, O.A., Stikhin, V.A. and Savchuk, V.M. (1984) Content of galanthamine in some 

Amaryllidaceae species of the flora of the Ukrainian SSR. Rastitel’nye Resursy, 20 (4), 566– 

568 (in Russian). 

Cherkasov, O.A., Tokhtabaeva, G.A., Margvelashvili, N.N., Maisuradze, N.I. and Tolkachev, 

O.N. (1988) The contents of galanthamine in some cultivars of Narcissus hybridus hort. 

Rastitel’nye Resursy, 24 (3), 414–420 (in Russian). 

Chimiko-Pharmazevtitschen Zavod (1959) A Method for Obtaining Galanthamine Hydrobromide. 

UK Patent No. GB 942 200; Bulgaria Patent Application No. BG 4404. 

Chimiko-Pharmazevtitschen Zavod (1963) A Method for Obtaining Galanthamine Hydrobromide. 

UK Patent No. GB 942 200. 

Cozanitis, D.A. and Rosenberg, P. (1983) Study of the analgesic effects of galanthamine, a 

cholinesterase inhibitor. Archives Internationales de Pharmacodynamie et de Thérapie, 266, 

229–238. 

Czollner, L., Frantsits, W., Kenburg, B., Hedenig, U., Frohlich, J. and Jordis, U. (1998) New 

kilogram-synthesis of the Anti-Alzheimer drug (–)-galanthamine. Tetrahedron Letters, 39, 

2087–2088. 

Erhardt, W. (1993) Narzissen. Eugen Ulmer Verlag, Stuttgart. 

Fitzgerald, D.B., Hartwell, J.L. and Leitner, J. (1958) Tumor-damaging activity in plant 

families showing antimalarial activity. Journal of the National Cancer Institute, 20, 763–774. 

Fuganti, C. (1975) The amaryllidaceae alkaloids. In: R.H.F. Manske (ed.), The Alkaloids, 

Vol. 15, Academic Press, New York.

272 M. Kreh 

Gorinova, N.I., Atanassov, A.I., Stojanov, D.V. and Tencheva, J. (1993) Influence of chemical 

composition of soils on the galanthamine content in Leucojum aestivum. Journal of Plant 

Nutrition, 16 (9), 1631–1636. 

Han, S.Y., Mayer, S.C., Schweiger, E.J., Davis, B.M. and Joullié, M.M. (1991) Synthesis and 

biological activity of galanthamine derivatives as acetylcholinesterase inhibitors. Bioorganic 

and Medicinal Chemistry Letters, 1 (11), 579–580. 

Hille, T., Hoffmann, H.R., Kreh, M. and Matusch, R. (1996) Verfahren zur Isolierung von 

Galanthamin. Germany Patent No. DE 195 09 663 A1. 

Jefferson-Brown, M. (1991) Narcissus. B.T. Batsford Ltd, London. 

Kington, S. (1989) The International Daffodil Checklist. The Royal Horticultural Society, 

London. 

Kreh, M. (1995) Galanthamin und andere Amaryllidaceen Alkaloide. Dissertation, Marburg 

University. 

Kreh, M., Matusch, R. and Witte, L. (1995) Capillary gas chromatography-mass spectrometry 

of Amaryllidaceae alkaloids. Phytochemistry, 38 (3), 773–776. 

Kreh, M., Müller, U. and Matusch, R. (1994) Bildung, Kristallstruktur und absolute 

Konfiguration von (–)-N-(Chloromethyl)galanthaminium-chlorid. Helvetica Chimica Acta, 

77, 1611–1615. 

Opitz, K., Lohmann Therapie-Systeme and Hefa-Frenon Arzneimittel (1991) Pharmazeutische 

Formulierung zur Behandlung des Alkoholismus. Germany Patent No. DE 40 10 079 A1. 

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the 2nd International Pharmacological Meeting, August 1963, Pergamon Press, New York, 

pp. 113–114. 

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003, 670519; Bulgaria Patent No. BG 590 302. 

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Obshchei Khimii, 17, 1216–1216 (in Russian). 

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Patent No. SU 145 583, 661 117, 610 503. 

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aus Pflanzen der Familie der Amaryllidaceen. Germany Patent No. DE 1 

193 061. 

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Zeitung, 100 (8), 43–43. 

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Springer Verlag, Berlin. 

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determination of galanthamine. Planta Medica, 56, 77–81.

10 Galanthamine production from 

Narcissus: agronomic and 

related considerations 

Rita M. Moraes 

INTRODUCTION 

Alzheimer’s dementia (AD) is an ultimately fatal degenerative disease of the central 

nervous system, affecting four million Americans. According to the Reagan 

Research Institute and the Alzheimer’s Association, families spend US$100 billion 

yearly caring for these patients. Furthermore, as the population ages, the number 

of Americans suffering from AD is expected to reach 6.2 million in the year 2015 

(Cutler et al., 1994). Galanthamine (1), an alkaloid found in species of the Amaryllidaceae, 

has been recommended for the treatment of AD (Davis, 1987) and is currently 

in phase III clinical trials. Positive results have prompted the trial’s sponsors, 

Shire Pharmaceuticals Group and Janssen Pharmaceutica Inc., to proceed with 

clinical development. As a selective and reversible inhibitor of acetylcholinesterase, 

this compound has the ability to cross the blood-brain barrier and act within the 

central nervous system (Mucke, 1997). 

O 

6 

O 

Galanthamine, 1 

3 

N 

OH 

1 

2 

In eastern Europe, galanthamine has long been used clinically as a recovery 

agent after anaesthesia (Schuh, 1976), and has been recently tested in a prodrug 

form to counteract the sedative, hypnotic and respiratory side effects of benzodiazepine 

therapy for schizophrenia (Snorrason, 1996). Other therapeutic uses include 

relief of jet lag (Davis, 1996), fatigue syndrome (Snorrason, 1994), male impotence

274 R.M. Moraes 

(Katz and Taneck, 1993) and nicotine (Moormann and Werner, 1997) and alcohol 

dependence (Opitz, 1996). Approval for the treatment of AD, together with its 

current uses, will create a strong and stable demand for galanthamine, even 

though other competitive anticholinesterase drugs are available. Recent estimates 

of the retail price of galanthamine are approximately US$50 000 per kilogram 

(Shieh and Carlson, 1994). Under present conditions, the projected demand for 

galanthamine would not be met by the current supply without significantly 

increasing its price. Most commercially available galanthamine is extracted from 

wild-harvested Leucojum aestivum in Bulgaria and Russia (Poulev et al., 1993). 

Research on the chemical synthesis of galanthamine has been aimed at total 

synthesis and also at the creation of more potent derivatives to minimise the 

dependence on natural resources (Bores and Kosley, 1996). Han et al. (1992) and 

Kosley et al. (1998) have prepared series of semi-synthetic derivatives. The adamantyl 

ester (2) is the most promising compound due to better affinity, selectivity 

for acetylcholinesterase and more favourable pharmacokinetics, thus reducing 

the amount of drug required to treat each AD patient. Research on total synthesis 

has improved galanthamine yields. However, the yield is still considered moderate 

(45 to 50%) and too low for economical commercial application (Czollner et al., 1998; 

Eichhorn et al., 1998). 

O 

O 

6 

O 

Adamantyl-ester galanthamine, 2 

In addition to galanthamine, there is great interest in several other Amaryllidaceae 

alkaloids due to their wide range of biological activities, including antiviral, antimalarial 

and anticancer properties (Gabrielsen et al., 1992; Likhiywitayawuid et al., 

1993). Pretazettine (11) is a highly cytotoxic compound against human lymphoid 

neoplasm. It has also been used in combination with DNA-binding alkylating agents 

in the treatment of the Rauscher leukaemia virus (Furusawa et al., 1978). Narciclasine 

(9) has anticancer, antimitotic and antiviral activity (Evidente et al., 1986; Pettit et al., 

1990). Lycorine (6) and lycoricidine possess plant growth-inhibiting properties and 

antiviral activity (Ieven et al., 1983). According to Jimenez et al. (1976), dihydrolycorine, 

haemanthamine (7), lycorine, narciclasine and pretazettine are protein inhibitors 

in eukaryotic organisms due to their binding to the 60S ribosomal subunit. 

Lycorine inhibits translation at the termination step (Vrijsen et al., 1985). 

3 

N 

OH 

1 

2

HCO 

3 

O 

O 

6 

O 

H 

N 

R 

OH 

Lycoramine 3, 

R=CH3 

DemethyI-Iycoramine 4, 

R=H 

O 

O 

Haemanthamine 7 

R1 

3 

O 

N 

1 

2 

OR2 

H 

NH 

OH 

Narciclasine 9, R 1=OH 

R=H 2 

Lycoricidine , R =R =H 

10 1 2 

Agronomic factors and galanthamine production 275 

OCH3 

OR2 

OR2 

Structural formulae of the Amaryllidaceae alkaloids 

At the National Center for Natural Products Research, the potential of Narcissus 

cultivars as sources of galanthamine and related alkaloids has been recognised, and 

research has been initiated on the agronomic factors that affect galanthamine content 

in them. Narcissus has two important advantages over Leucojum aestivum: firstly, 

O 

O 

O 

O 

O 

O 

Caranine 5, 

R=H 

Lycorine 6, 

R=OH 

H 

N 

HO 

H 

O 

H 

H 

O 

R 

Hippeastrine 8 

OCH3 

OH 

O 

Pretazettine 11 

H 

N 

H 

H 

N 

OH


bulbs of many Narcissus cultivars are available in commercial quantities, offering 

the possibility of establishing large-scale cultivation for medicinal purposes in a 

short time; secondly, a comprehensive body of information already exists regarding 

narcissus propagation, physiology, breeding and cultivation for flower production 

(e.g., Rees, 1992; Hanks, 1993). In this chapter, the results of studies on 

planting density and depth, bulb size, and alkaloid distribution within the bulb 

and at different stages of bulb development, are reported. Agronomically important 

aspects of narcissus production for medicinal purposes will also be discussed. 

AMARYLLIDACEAE FOR THE SUPPLY OF GALANTHAMINE 

The present commercial sources for galanthamine are wild populations of Leucojum 

and Galanthus. The sourcing and cultivating of medicinal species are requirements 

for the progression of some natural products into pharmaceuticals. 

Treatment of large patient populations requires a reliable supply of the active 

compound at an affordable price. Therefore, the cultivation of medicinal plants is 

a growing segment of the pharmaceutical industry, and there is an urgent need 

for high-quality biomass from non-wild sources to benefit consumers and to protect 

the environment. Palevitch (1991) has reported the advantages of medicinal 

plant cultivation over collection from the wild. These are: quality products with 

botanical source assurance, a stable market for consumers with increasing availability 

of plant material, and less fluctuation in supply. 

Extensive surveys seeking richer galanthamine sources have been carried out on 

most of the Old World taxa of the Amaryllidaceae, but New World amaryllids, 

especially the numerous neotropical genera, have rarely been examined. Poulev 

et al. (1993) reported that Phaedranassa megistophylla, a species native to Peru, contains 

7.4% galanthamine on a dry weight basis. It appears to be a very promising 

source, since this concentration is several times higher than the highest concentration 

reported for any other source of galanthamine. 

Galanthamine is not only found in Amaryllidaceae sensu stricto: low concentrations 

of galanthamine have been detected in the closely related families of 

Agavaceae, Haemodoraceae and Hypoxidaceae using an enzyme immunoassay 

procedure (Poulev et al., 1993). Using a different method (Bastos et al., 1996), most 

of the findings of Poulev et al. (1993) have been confirmed, and in a survey of 

Narcissus, more than 80 taxa were analysed. Extracts from dormant bulbs of 

Narcissus ‘Inglescombe’ had the highest galanthamine content, 173.7 mg per 100 g 

of dry weight (0.17%). A wide genotypically fixed variation in alkaloid content 

among cultivars was found (Bastos et al., 1996; Moraes-Cerdeira et al., 1997b). 

ALKALOID CONTENT OF DIFFERENT PARTS OF THE BULB 

The importance of galanthamine as a therapeutic drug, and the biological value of 

other narcissus alkaloids, prompted the study of the distribution of several alkaloids 

in specific bulb parts (Moraes-Cerdeira et al., 1997a). Rees (1969) described 

the narcissus bulb as a complex branching system composed of several ‘bulb units’. 

Each bulb unit consists of a shoot apex enclosed by bulb scales and leaf bases acting


Table 10.1 Distribution of Narcissus alkaloids in different bulb parts 

Part of bulb Weights (g) % total Narcissus alkaloids (mg/100 g dry weight) 

Fresh Dry 

dry wt. 

Total 1a 2 3 4 5 6 7 

Basal plates 166.7 47.3 8.3 461.2 168.5 40.9 37.1 10.8 113.7 75.6 14.6 

Outer scales 484.3 186.8 32.9 148.5 59.8 16.0 22.0 6.3 25.0 18.7 trace 

Inner scales 738.1 268.6 47.4 196.7 78.8 9.9 9.9 2.4 63.2 32.5 trace 

Leaves 85.7 16.2 3.0 455.3 132.7 7.7 1.3 20.1 226.5 67.0 trace 

Flowers 87.9 28.3 4.9 223.6 68.3 2.7 trace 30.5 73.0 49.1 trace 

Bulbil 77.3 19.8 3.5 258.7 105.5 6.2 4.6 13.2 85.6 43.6 trace 

Note 

a 1 = galanthamine, 2 = lycoramine, 3 = N-demethyl-lycoramine, 4 = caranine, 5 = haemanthamine, 

6=lycorine, and 7=hippeastrine. 

as storage organs. The scales are held together concentrically by the base plate, 

a solid, stem-like tissue at the base of the bulb. The shoot apices initiate scales, 

leaves, stems, flowers and new bulb units. 

The study of Moraes-Cerdeira et al. (1997a) revealed that the basal plate has 

the highest content of galanthamine and of the other dibenzofuran alkaloids, 

lycoramine and N-demethyl-lycoramine (Table 10.1). Galanthamine concentrations 

were higher in the inner bulb parts. Lycorine, a highly toxic, norpluvine-derived 

alkaloid, is similar in its distribution to galanthamine, with a higher concentration 

in the inner parts. Galanthamine, lycoramine and N-demethyl-lycoramine have the 

same biogenetic precursor, but lycoramine and N-demethyl-lycoramine are found 

primarily in the scales. Therefore, no pattern of alkaloid distribution was found 

relating to organ specificity and biogenetic origin. 

Meristematic tissues contributed one-third (33.5%) of the total alkaloids of the 

bulbs. Scales had 66.5% of the total content in 80% of the total dry weight of the 

bulb (Table 10.1). The active growth of narcissus bulbs leads to an increased ratio 

of meristematic tissue to scale (storage tissue) through growth that depletes the 

starch-containing tissues. Therefore, treatments stimulating growth enrich galanthamine 

and alkaloids like lycorine (6), haemanthamine (7) and the caranine (5) 

content of the biomass, but it may also decrease the content of lycoramine (3) and 

N-demethyl-lycoramine (4). 

Data from our laboratory on different growth stages of Narcissus ‘Inglescombe’ 

(Figure 10.1) suggested that plants produced more galanthamine during the 

growing period, between emergence and anthesis, reaching as high as 287 mg per 

100 g dry weight (C.L. Burandt, Jr. et al., unpublished data). Galanthamine content 

decreased after anthesis to 173 mg per 100 g dry weight, a stage marked by scale 

growth and leaf senescence. As starch content increased, galanthamine appeared 

to become diluted by a metabolic shift towards starch production. Both studies 

described above indicated that agronomic factors which stimulate cell growth may 

enhance galanthamine content by 50% or more on a dry weight basis. Therefore, 

narcissus biomass for galanthamine production should be harvested in April and 

May at full foliage in an 18-month growing cycle. Alternatively, it may be even 

more economical to lift bulbs after a 24-month crop cycle, then force bulb emergence 

and growth by storing the bulbs in a controlled environment at high humidity 

and low temperature.


Galanthamine content 

mg /100g dry weight 

300 

250 

200 

150 

100 

50 

0 

AGRONOMIC FACTORS AFFECTING DRUG YIELD 

Genetic and environmental factors strongly influence biomass growth and its 

content of secondary compounds. To achieve maximum yields of any active compound 

such as galanthamine, optimum agronomic practices should be established. 

The agronomic practices include the identification of high-yielding cultivars, 

optimum growing conditions (climate, soil type, planting density, fertilisation and 

irrigation), pest management (weeds, insects and disease), and appropriate 

mechanisation for cultivation, harvesting, drying and grinding. 

As mentioned above, a survey of more than 80 Narcissus taxa indicated that 

Narcissus ‘Inglescombe’ had the highest galanthamine content (Bastos et al., 1996). 

However, the current supply of ‘Inglescombe’ bulbs is small and requires a mass 

propagation programme to increase stocks for extensive cultivation. 

Propagation 

I II III IV V VI VII VIII IX X XI XII 

Developmental Stage 

Figure 10.1 The effect of developmental stages of Narcissus ‘Inglescombe’ on galanthamine 

content during the 1994 growing season (unpublished data): 

I, pre-emergence (1 February); II, emergence, pre-bud stage (26 February); 

III, post-emergence, early bud stage (10 March); IV, anthesis (23 March); 

V, post-anthesis, ‘fruiting’ (25 April); VI, senescing (2 June); VII, senescent 

(6 July); VIII, senescent (28 July); IX, senescent (29 August); X, root 

re-growth (4 October); XI, shoot re-growth (4 November); XII, underground 

growth (10 December). 

The natural rate of narcissus bulb multiplication is low: the average rate of 

increase is 1.6 per annum, or 1 to 1000 in 16 years (Rees, 1969). Faster methods of 

propagation are available, such as twin-scaling (Hanks and Rees, 1979; Fenlon


et al., 1990), chipping (Flint and Alderson, 1986), and micropropagation (Squires 

and Langton, 1990). Mechanised chipping (Hanks, 1989) is a large-scale propagation 

method for abundant low-priced stocks of narcissus. In terms of achieving 

high numbers of propagules, the micropropagation procedure can reach 2.5 million 

in five years, much greater than the numbers produced by twin-scaling or chipping. 

However, due to the high cost of the facilities and the intensive labour 

required, micropropagation remains complementary to chipping and twin-scaling 

in the ornamentals industry. Currently, micropropagation is only used to a very 

limited extent on narcissus, to multiply small stocks of elite, virus-free lines and 

cultivars, and for germplasm propagation in breeding programmes. A combination 

of available propagation methods may be the wisest strategy to increase narcissus 

bulb stocks for galanthamine production. However, the magnitude of the losses, 

economic and otherwise, that AD causes to patients, families and society, may 

justify the high cost of micropropagating narcissus bulbs for medicinal purposes. 

Therefore, micropropagation may be more affordable in the pharmaceutical 

industry than in the ornamentals industry. If so, micropropagation could rapidly 

increase propagule stocks of high-yielding cultivars such as ‘Inglescombe’, improving 

biomass quality for better extractable yields. 

Growth and biomass yield 

As long as the supply of galanthamine relies on natural sources, there will be 

an increasing need to optimise biomass production and extraction to achieve 

maximum yields. A research programme on galanthamine supply from natural 

sources has both short- and long-term strategies. The short-term strategy includes 

a procedure with three fundamental steps, which will indicate the feasibility of 

pilot-scale production system. The first step consists of the identification of taxa 

found in abundance that could become a reliable source for establishing a production 

system. The second step is the optimisation of growing conditions and 

agronomic practices to maximise field production of the abundant source. The 

third step constitutes the development of a large-scale extraction and separation 

method to improve the yields of galanthamine isolated. 

The long-term strategy consists of selecting high-yielding Narcissus taxa and 

other Amaryllidaceae, maintaining germplasm collections, mass propagating elite 

plants, and field studies to enhance yields in an already established production 

system. At the National Center for Natural Products Research, four widely available 

Narcissus cultivars, ‘Geranium’, ‘Mount Hood’, ‘Cheerfulness’ and ‘Ice Follies’, 

have been identified and evaluated for agronomic factors affecting their 

yield. The results indicated that planting depth, bulb size, planting density and the 

removal of flower buds did not influence drug content. However, these factors did 

influence bulb growth, and therefore drug yield per unit of cultivation area (Table 

10.2) (Moraes-Cerdeira et al., 1997b). ‘Cheerfulness’ and ‘Geranium’ exhibited 

good growth but low galanthamine content, thus showing little promise for drug 

production. 

Planting density is a complex issue. Under dense plantings, the decreased 

growth rate reduces the yield per plant, but the biomass yield per unit area of land 

increases (Rees et al., 1973). Galanthamine content is unaffected by high-density 

plantings. Therefore, depending on the growth increase, planting density may


Table 10.2 The effect of planting depth on bulb growth and galanthamine yield of four 

Narcissus cultivars 

Cultivar Growth % Galanthamine (mg/100 g DW) 

Note 

a Least significant difference at p


In order to reach maximum yields of galanthamine, bulb lifting might be early in 

the spring, when meristematic activity is at a maximum. Harvesting in early spring 

may reduce herbicide application and adulteration problems caused by weeds 

in heavily infested fields. On the contrary, lifting dormant bulbs may facilitate 

biomass-handling processes, but it will require post-harvest treatments to promote 

bulb sprouting to enhance galanthamine yield. 

CONSIDERATIONS AND PERSPECTIVES FOR SHORT-TERM 

GALANTHAMINE PRODUCTION FROM NARCISSUS 

The world’s largest producer of narcissus is the UK, with a production area of 

about 4500 ha (Rees, 1992). Other major producers include the Netherlands, US 

and Poland (Hanks, 1993). Narcissus ‘Ice Follies’ is a fast growing and popular 

cultivar, ranking third in flower bulb sales according to the International Bulb 

Society, Pasadena, California. In the Netherlands, the cultivated area of Narcissus 

‘Ice Follies’ in 1998/99 was given as 101 hectares (Anon., 1999). However, no equivalent 

data have been found on production areas of ‘Ice Follies’ in the other major 

producer countries. Therefore, we have no information on the overall commercial 

availability of bulbs of this cultivar, but as it is among the best selling cultivars one 

can expect a reasonable amount of available stock with which to initiate pilot-scale 

production. Additionally, ‘Ice Follies’ contains reasonable amounts of galanthamine: 

the average content extracted from dormant bulbs is 70 mg per 100 g dry weight 

(Moraes-Cerdeira et al., 1997b). 

The recommended daily dose of galanthamine to treat an AD patient varies 

between 30 and 50 mg (Dal-Bianco et al., 1991). Therefore, a single patient would 

need around 20 g of galanthamine per year. Today, there are 4 million AD 

patients in the US alone. To treat 30% of these patients with galanthamine hydrobromide, 

24 000 kg of the compound would be required per year. Aiming for a 

30% market is considered a standard goal for pharmaceutical companies to invest 

into a new product. 

Under Mississippi climatic conditions, Narcissus ‘Ice Follies’ produced 14 300 kg 

of bulbs per hectare in one growing cycle (Moraes-Cerdeira et al., 1997b), representing 

a growth increase of 70%. Based on this example of bulb production and 

using commercial scale drug isolation of 65% efficiency, the cultivated area necessary 

to treat 30% of the US patients in one year would be 10 018 ha. This estimated 

area reflects the bulb yield of Narcissus ‘Ice Follies’ grown in Mississippi under suboptimal 

climatic conditions (Anderson, 1989) and 36.82% dry weight of the fresh 

bulbs with 70 mg of galanthamine per 100 g of dry biomass. Under the UK conditions, 

narcissus bulbs are typically planted at a rate of around 17.5 t/ha, and a 

weight increase of at least 120% would be expected for an average variety after two 

years, a total yield of about 38.5 t/ha (Hanks, 1993). Such high-density plantings 

would be a suitable growing system for a cheap cultivar with a low yield of galanthamine, 

and on this basis an area of 13 636 ha would be needed to produce the 

same amount of galanthamine as in the example above for ‘Ice Follies’ growing in 

Mississippi, but for a two-year demand to treat 30% of AD patients. Using chipped 

bulbs (eight chips per bulb) and planting 2 t of chipped bulbs per ha, a reasonable 

expectation would be a nine-fold increase in weight and number of bulbs after two


years, a yield of 18 t/ha (Langton and Hanks, 1993). This would be a more suitable 

growing system for a high galanthamine content cultivar which was expensive, 

and in short supply. An area of 7164 ha would be needed to equal the previous 

production of galanthamine for a two-year drug supply to treat 30% of the US 

Alzheimer patients using high yielding cultivars such as Narcissus ‘Inglescombe’ 

(174 mg per 100 g of dry weight at bulb dormant stage). Cultivation of high yielding 

bulbs would decrease all production requirements and costs (plantation size, 

handling, storage, extraction and waste products). Furthermore, harvesting 

‘Inglescombe’ during active growth (284 mg per 100 g dry weight) would yield 

63% more galanthamine than dormant bulbs and would decrease costs even more. 

Another strategy to reduce the cultivation area would be the semi-synthetic 

approach, extracting galanthamine analogues (lycoramine, demethyl-lycoramine, 

narwedine and demethylnarwedine) and chemically converting these to galanthamine. 

Following the first promising results of galanthamine in clinical trials, 

many semi-synthetic derivatives of galanthamine were synthesised. A series of 

carbamates of 6-demethylgalanthamine were found 1000 times more potent than 

galanthamine as cholinesterase inhibitors (Bores and Kosley, 1996). Several esters 

were also prepared and among these, the adamantyl ester (2) is the most promising 

derivative. In addition, the adamantyl ester has greater selectivity and a better 

oral pharmacokinetic profile, and thus a higher oral therapeutic index, compared 

with galanthamine. 

The economically viable production of galanthamine, directly or by semisynthesis 

from natural resources for pharmaceutical purposes, calls for the co-ordinated 

contribution of many scientists in a multidisciplinary effort. Treatment of 

AD patients with galanthamine will require the production of large quantities of 

narcissus biomass of high galanthamine content. Substantial research is needed to 

define the most favourable agronomic practices and to optimise extraction and 

isolation of the pure compound. 


Special thanks are due to James D. McChesney for his support and guidance 

during this research on galanthamine production from narcissus. My gratitude to 

the dedication and views of my collegues and collaborators on our galanthamine 

work, Jairo K. Bastos, Charles L. Burandt Jr., Julie Mikell, N.P. Dhammika 

Nanayakkara and Thomas Sharpe. I am especially indebted to Joseph Atkins, 

Allison Best, Camilo Canel, Hala ElSohly, Jeffrey Krans and Larry Walker for 

their helpful discussions on this chapter. Thanks to my sons Cesar, Marcos and 

my mother Maria Martha for their love. 

This chapter is dedicated to the memory of my loving father and great 

agronomist Leo Gomes de Moraes. 

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11 Extraction and quantitative analysis 

of Amaryllidaceae alkaloids 

N.P. Dhammika Nanayakkara and Jairo K. Bastos 

INTRODUCTION 

Amaryllidaceae alkaloids represent a diverse class of natural bases that occur in 

different species of the family Amaryllidaceae. Owing to their wide spectrum of 

useful biological activities, these compounds have attracted the attention of both 

chemists and biologists. Many Amaryllidaceae alkaloids have been isolated and 

identified, and a number of methods to quantify them have been developed. 

Galanthamine, a common alkaloid in this family (Figure 11.1), has shown cholinesterase 

inhibitory activity and is currently undergoing clinical trials for the 

treatment of Alzheimer’s disease. Another member of this family, pancratistatin, 

has shown a highly characteristic differential cytotoxicity profile against a panel of 

human cancer cell lines and strong activity against RNA viruses. This compound is 

undergoing development towards human clinical trials. In an effort to find new 

sources of galanthamine (Tanahashi et al., 1990; Poulev et al., 1993; Bastos et al., 

1996) and pancratistatin (Pettit et al., 1995a,b), several species of Amaryllidaceae 

have been examined. 

In the early years, quantitative analytical studies of Amaryllidaceae alkaloids 

were carried out using gravimetric methods. During the last four decades, several 

other analytical procedures have been described. Most of these methods were 

developed to quantify galanthamine in biological samples and natural sources. 

Several sensitive, practical methods to analyse most common alkaloids of this class 

in natural sources, simultaneously and quantitatively, were also reported. In this 

chapter, procedures published over the last four decades are reviewed. 

MeO 

O 

H 

N 

Me 

OH 

O 

O 

OH 

Galanthamine Pancratistatin 

Lycorine 

HO 

H 

O 

Figure 11.1 Chemical structures of Amaryllidaceae alkaloids with important biological 

activities. 

OH 

H 

NH 

OH 

OH 

O 

O 

HO 

H 

OH 

H 

N

Analysis of Amaryllidaceae alkaloids 287 

EXTRACTION OF AMARYLLIDACEAE ALKALOIDS 

In quantitative analytical studies, alkaloids are usually extracted using the traditional 

method. Plant material is typically dried and ground prior to extraction. 

However, fresh plant materials are often used for this purpose, the water 

content of which fluctuates with season, age, tissue type and storage conditions. 

For this reason, quantitative data generated for fresh plant samples collected 

and processed under different conditions may not be reliable for comparative 

purposes. Traditionally, dried ground plant material, or fresh plant material 

after maceration, is extracted with ethanol or a dilute acid solution. To remove 

lipophilic non-alkaloids, the dilute acid extract is directly partitioned with an 

organic solvent, whereas the ethanol extract must be evaporated and reconstituted 

in dilute aqueous acid prior to partitioning. The use of fresh plant material 

can present problems in the evaporation process, as large amounts of water and 

partially water-soluble polysaccharides, especially in bulbs, make evaporation of 

the ethanol extract difficult. After the removal of ethanol, the concentrated 

extract is diluted with an acid solution and partitioned with an organic solvent. 

The polysaccharides can also interfere with solvent extraction processes, tending 

to make thick emulsions and causing incomplete extraction of non-alkaloids 

and alkaloids in the subsequent steps. After the removal of lipophilic constituents, 

the aqueous layer is basified and alkaloids are extracted into an organic 

solvent. 

In quantitative studies, where small samples are involved, problems due to 

emulsions in the solvent extraction process can be overcome by centrifugation 

(Davey et al., 1998; Bastos et al., 1996). However, a thick interface between organic 

and aqueous layers may cause incomplete removal of the target compounds. The 

error caused by this can be corrected by the addition of an internal standard at the 

beginning of extraction, and by carrying out recovery studies for the substrates 

and the internal standard (Claessens et al., 1983; Tencheva et al., 1987; Bastos 

et al., 1996). 

Queckenberg and Frahm (1994) developed a supercritical fluid extraction 

procedure to remove the alkaloid fraction selectively from methanol extracts. The 

re-extraction of dried methanol extract of Amaryllidaceae plants with supercritical 

nitrous oxide (N 2 O) in the presence of a modifier (methanol containing ammonia 

gas) efficiently removed the alkaloids, which were subsequently analysed by chromatographic 

methods (Queckenberg and Frahm, 1993; Queckenberg et al., 1996). 

Zhu et al. (1993) developed a supercritical fluid extraction method to isolate 

lycorine from bulbs of Amaryllidaceae, and optimum conditions for the separation 

were determined. 

The procedure for the extraction of alkaloids (and their metabolites) from biological 

fluids or tissues is somewhat similar to that used for plant materials. However, 

due to the complex nature of the biological samples, more steps are involved 

(Bickel et al., 1991a,b; Tencheva et al., 1987). Recently, Bores et al. (1996) used a 

solid phase extraction procedure to separate galanthamine and its metabolite, 

6-O-demethylgalanthamine, from biological samples. This procedure involves 

the application of biological fluids containing test compounds to reversed-phase 

cartridges and selective elution of the compounds of interest with appropriate 

solvents.

288 N.P.D. Nanayakkara and J.K. Bastos 

In many instances, the separation method can be simplified based on the selectivity 

of the detection mode. In studies where highly specific detection techniques 

(e.g., immunoassay) were used, minimum sample processing was necessary (Tanahashi 

et al., 1990; Poulev et al., 1993). 

QUANTITATIVE ANALYSIS OF AMARYLLIDACEAE ALKALOIDS 

Gas chromatography 

Packed column-gas chromatography (PC-GC) 

In early studies, the feasibility of gas chromatography for the analysis of alkaloids 

was explored using packed columns. Lloyd et al. (1960) analysed several underivatised 

Amaryllidaceae alkaloids by PC-GC, and single component sharp peaks for 

these compounds were observed in chromatograms. Yamaguchi et al. (1962) 

reported the detection of galanthamine, lycoramine and tazettine by PC-GC 

under similar conditions. However, this method was not applicable to non-volatile 

alkaloids, such as lycorine, and other hydroxylated alkaloids showed significant 

peak tailing, especially with polar liquid surfaces. 

Takagi et al. (1968a) described the GC analysis of twelve Amaryllidaceae alkaloids, 

using several commercially available polar and non-polar stationary phases. 

The test compounds were converted to their trimethylsilyl derivatives prior to 

analysis. The trimethylsilyl derivatives of these compounds showed symmetrical 

and relatively sharp peaks, even for non-volatile alkaloids such as lycorine. 

Multiple peaks for lycorenine were observed under these conditions, presumably 

due to decomposition. These authors later applied this method to determine the 

quantitative changes in lycoramine and galanthamine content in Lycoris radiata 

through a period of one year (Takagi et al., 1968b). In a subsequent study (Takagi 

et al., 1969), the lycorenine content in L. radiata was determined quantitatively by 

converting it into tetrahydrohomolycorenine and analysing the trimethylsilyl 

ether derivative by PC-GC. 

Packed column-gas chromatography-mass spectrometry (PC-GC-MS) 

Millington et al. (1972) analysed the alkaloid fraction of Crinum glaucum after trimethylsilyl 

derivatisation by PC-GC-MS in electron impact mode. In addition to the 

identification of ambeline and lycorine, several new alkaloids were recognised. In a 

subsequent report (Onyiriuka and Jackson, 1978), the same group used PC-GC-MS 

in electron impact mode, in combination with other mass spectral data and 

nuclear magnetic resonance (NMR) spectroscopic information, to detect and 

propose chemical structures for several new compounds in alkaloid fractions of 

Crinum ornatum and C. natans. 

Capillary gas chromatography (CGC) 

A CGC method coupled with nitrogen phosphorus detection was also used for 

the quantitative analysis of Amaryllidaceae alkaloids in plant extracts (Bastos 

et al., 1996). In this study, baseline resolution for eleven reference alkaloids


and an internal standard was achieved using a 15 m capillary column within a 

15 minute run time. Good linearity of response was observed for all the reference 

compounds in a concentration range of 8–500 µg/ml, and concentrations 

around 10 µg/ml could be quantified reproducibly using this method. Quantitative 

analysis was carried out using 500 mg of dried plant material, and this 

method was used to quantify alkaloids in different plant parts (Moraes- 

Cerdeira et al., 1997a), and to evaluate the galanthamine content of several 

Narcissus cultivars grown under experimental conditions (Moraes-Cerdeira 

et al., 1997b). 

Capillary gas chromatography-mass spectrometry (CGC-MS) 

Kreh et al. (1995a) studied the application of CGC-MS in the detection and identification 

of underivatised Amaryllidaceae alkaloids. Fifteen reference alkaloids 

were subjected to CGC-MS in electron impact or chemical ionisation mode, and 

their stability under column conditions was studied. All but two compounds were 

found to be stable, with haemanthamine and lycorenine undergoing partial 

decomposition during analysis. Derivatisation with trimethylsilylating agents failed 

to improve either the sensitivity or the resolution of the analysis. Using this 

method, they were able to detect and propose the structures of several new 

compounds in Narcissus ‘Carlton’ (Kreh et al., 1995b). 

High performance liquid chromatography (HPLC) for the 

determination of Amaryllidaceae alkaloids in natural sources 

High performance liquid chromatography (HPLC) 

Westwood et al. (1981) investigated the feasibility of coupling HPLC with a circular 

dichroism (CD) spectrometer to analyse optically active compounds with suitable 

chromophores selectively. By using stopped-flow techniques, full CD spectra of 

optically active compounds were recorded. An alkaloid fraction of Crinum glaucum was 

chromatographed using a reversed-phase C-8 column and a mixture of methanolwater 

with a trace of ammonia as the solvent. One of the major compounds in the 

extract, ambelline, was detected through a comparison of the on-line recorded CD 

spectrum with that obtained under standard conditions. 

Evidente et al. (1983) developed a rapid quantitative analytical procedure for 

lycorine using a C-18 reversed-phase column and acetonitrile:0.01 M ammonium 

carbonate (47:53) as the solvent. Lycorine was analysed as the sulphate salt, and 

was detected using an ultra-violet (UV) detector at 290 nm. The detection limit for 

lycorine was 5 ng. These authors applied the method to quantify lycorine in crude 

acid extracts of bulbs and leaves of Sternbergia lutea. Davey et al. (1998) adapted this 

system under semi-preparative and analytical conditions to purify and quantify 

lycorine from acid extracts of Crinum asiaticum. 

Bruno et al. (1985) described an HPLC method to determine lycorine and 

tazettine in the bulbs of Narcissus tazetta. The alkaloid fraction was separated on a 

LiChrosorb-CN ® column using methanol:water (25:75) containing 1% Pick 

reagent as the mobile phase. Compounds were detected by fluorescence, using an


excitation wavelength of 290 nm and an emission wavelength of 320 nm. The 

detection limit was less than 5 ng/ml. 

Könükol and Sener (1992) described a reversed-phase HPLC procedure to 

quantify lycorine and crinine simultaneously in bulbs of Pancratium maritimum. 

A reversed-phase C-18 column and a mobile phase containing a mixture of 

chloroform:methanol (90:10), with UV detection at 292nm, were used in this 

procedure. 

Another reversed-phase method was reported by Sellés et al. (1997), who quantitatively 

evaluated galanthamine, N-formylgalanthamine, haemanthamine and 

tazettine in wild populations and tissue cultures of Narcissus confusus. They used a 

C-18 column and a mobile phase consisting of water (containing phosphoric acid, 

pH 3) (solvent A) and acetonitrile (solvent B) in a 60:40 ratio. Peak broadening 

and tailing were minimised by the addition of 10 and 12 mM of octanesulphonic 

acid to solvents A and B, respectively. Compounds were detected using a diode 

array UV detector operating at 280 nm. 

High Performance Liquid Chromatography (HPLC) – Thin Layer 

Chromatography (TLC) 

Queckenberg and Frahm (1993) developed a procedure wherein reversed-phase 

HPLC was coupled with automated multiple development (AMD) on normal 

phase silica to detect and quantify Amaryllidaceae alkaloids in natural sources. At 

first, conditions such as stationary phase, base solvent, modifier, buffer (pH and 

concentration), gradient profile, flow rate, temperature and sample size and solvent 

were optimised to achieve the best HPLC resolution for 20 reference Amaryllidaceae 

alkaloids. Under optimum HPLC conditions, the eluent from the HPLC 

column was applied to silica TLC plates and subjected to AMD. Compounds on 

the developed TLC plate were detected and quantified by different spectroscopic 

and chemical methods. The UV spectra of individual bands were used in combination 

with retention data to confirm the identity of each compound. Densitometric 

evaluation by a TLC scanner in the absorption/reflectance mode was used to 

quantify individual bands. The detection limit for tazettine by this method was 

0.2 ng. These authors applied the method to re-investigate the alkaloid extract of 

Amaryllis belladona (Queckenberg et al., 1996). This led to the identification of nine 

alkaloids, in addition to five previously known from this species, and three more 

were tentatively identified. 

High Performance Liquid Chromatography – Mass Spectrometry (HPLC-MS) 

By coupling MS and HPLC, Eckers et al. (1980) analysed an alkaloid fraction of 

Crinum glaucum. The extract was separated by reversed-phase HPLC using an 

ODS 5 µ Spheresorb ® column and water:acetonitrile:ammonia (20:79.7:0.3) as 

the mobile phase. The eluent was subjected to MS with a moving belt interface, 

and total ion current in electron impact (EI) mode was recorded. Two alkaloids, 

lycorine and ambelline, were identified on the basis of their EI spectra and 

retention times. Another compound, criglaucine, of uncertain structure, was also 

detected.


High performance liquid chromatography (HPLC) for the 

determination of galanthamine and its metabolites in 

biological fluids and tissues 

High performance liquid chromatography (HPLC) 

In addition to the analysis of plant material, several HPLC methods were developed 

to screen galanthamine and its metabolites in biological fluids. Claessens et al. 

(1983) initially reported an HPLC method for the quantitative determination of 

galanthamine in biological fluids using a normal phase silica column with hexanedichloromethane 

containing ethanolamine as the mobile phase. The minimum 

detectable concentration was 5 ng/ml, and the standard deviation varied between 

18.9 and 2.5% for the concentration range 10–100 ng/ml. Subsequently, a reversedphase 

HPLC method was described, using a C-18 column and acetonitrilepentanesulfonic 

acid buffer in water as the solvent (Claessens et al., 1988). In this 

method, body fluids were pre-processed by a preparative isotachophoresis prior to 

chromatographic analysis. Satisfactory recoveries, excellent resolution and good 

sensitivity were achieved. In both these procedures, a UV detector operating at 

235 nm was used to detect the test compounds. 

Tencheva et al. (1987) and Tencheva and Budevski (1987) developed a 

reversed-phase HPLC method for the quantitative determination of galanthamine 

and its metabolites, epigalanthamine and galanthaminone, in human body fluids. 

They used a C-8 column and a methanol:water (40:60) mobile phase modified 

with dibutylamine adjusted to pH 7 with 85% phosphoric acid. Codeine was used 

as an internal standard. The test compounds were detected by fixed-wavelength 

UV detector (280 nm). The detection limit was 0.05 µg/ml. This method was also 

subsequently used to quantify galanthamine in several native populations of 

Leucojum aestivum (Gorinova et al., 1993). 

A somewhat similar HPLC method was reported by Bickel et al. (1991a) to 

determine galanthamine and epigalanthamine in plasma and tissues of mice. 

A reversed-phase C-8 column and a mobile phase containing acetonitrile, tetrahydrofuran, 

water and di-n-butylamine at pH 7, adjusted with 85% phosphoric 

acid, were employed in this procedure. Codeine was used as an internal standard, 

and a fluorescence detector, set to 290 and 320 nm for excitation and emission 

frequencies, respectively, was used to detect the test compounds. 

High performance liquid chromatography – mass spectrometry (HPLC-MS) 

An HPLC-MS procedure for the determination of galanthamine and its metabolite, 

6-O-demethylgalanthamine in biological samples was described by Bores 

et al. (1996). In this procedure, a reversed-phase HPLC system was utilised with a 

mass spectrometer operating both in the multiple reaction monitoring mode (using 

a heated nebuliser interface) and in the selected ion monitoring mode (with an 

electrospray interface). 

Thin layer (TLC) and paper chromatography (PC) 

Several thin layer and paper chromatographic methods have been developed 

for the quantitative and qualitative analysis of Amaryllidaceae alkaloids.


Sandberg and Michel (1963) analysed the alkaloid profiles in different parts 

of Pancratium maritimum collected from different geographic areas using a 

two-dimensional TLC procedure. Separation of the alkaloids was carried out 

on 10 × 10 cm silica gel plates using mobile phases diethylether: methanol: 

diethylamine (85:10:5) and chloroform:methanol:diethylamine (92:3:5) in 

the two directions. Iodoplatinate reagent was used for visualisation. A total of 

fifty-two alkaloids, ranging from 14 to 37 per specimen, were detected in 

these samples, and their relative percentages were estimated. Five of them 

were isolated and identified as lycorine, haemanthidine, tazettine, vittatine 

and hordenine, and four more new compounds were also partially characterised. 

TLC on alumina was employed by Asoeva and Vergeichik (1967) to separate 

and quantify alkaloids in Galanthus krasnovii. Benzene:ethanol was used as the 

mobile phase, and the content of galanthamine and four other unidentified alkaloids 

were determined. Leifertova and Brazdova (1967) analysed G. nivalis collected 

from different regions by TLC and PC, and seven compounds, including galanthamine, 

lycorine and tazettine, were detected. 

A quantitative and qualitative study of the alkaloid composition of wild and 

introduced Leucojum aestivum populations was carried out by Stefanov (1977). 

Alkaloid fractions were separated by TLC on silica gel using methanol:diethylether:diethanolamine 

(5:90:5) as the mobile phase, and six compounds (galanthamine, 

galanthaminone, lycorine, lycorenine, nivalidine and an unidentified 

alkaloid) were determined quantitatively. 

The alkaloid fraction of common snowdrop bulbs (Galanthus nivalis) was studied 

by TLC on silica gel using chloroform:acetone:diethylamine (50:40:10), n-hexane: 

chloroform:acetone:diethylamine (80:25:30:5) and toluene:acetone:chloroform: 

diethylamine (45:25:25:5) as mobile phases (Kalashnikov, 1969). Dragendroff 

reagent was used as the visualising agent and six alkaloids, including galanthamine 

and lycorine, were detected. 

Wurst et al. (1980) developed a quantitative TLC procedure to determine the 

galanthamine content in extracts of Leucojum aestivum. The alkaloid fraction was 

separated by TLC on silica gel using diethylether:methanol:diethylamine 

(80:15:5) as the mobile phase, and the compounds were detected using a TLC 

scanner operating at 288 nm. The detection limit for galanthamine by this procedure 

was about 0.2 µg, and good linearity of response and reproducibility were 

obtained. 

Hong et al. (1981) developed TLC and PC procedures to identify galanthamine 

and lycoramine. In TLC, the best results were obtained on alumina by using either 

cyclohexane:chloroform:diethylamine (5:4:1) or benzene:ethyl acetate:diethylamine 

(7:2:1) as mobile phases. In PC, the paper was treated with a buffer at 

pH 4.4, saturated with water vapour prior to use, and chloroform was used as 

the solvent. 

Dobronravova et al. (1982) studied the chromatographic behaviour of halide 

salts of some alkaloids on alumina. TLC of hydrochloride salts of lycorine 

and galanthamine showed two spots after spraying with Dragendroff reagent, 

one major and the other near the start, presumably due to decomposition. 

The sensitivity of detection for these compounds by Dragendroff reagent was 

500 µg.

Immunoassays 


Immunoassay provides a sensitive and specific tool for many analytical tasks. 

Tanahashi et al. (1990) developed a radioimmunoassay procedure for quantifying 

galanthamine. The antiserum was raised in rabbits against a conjugate of galanthamine-2-O-hemisuccinate-bovine 

serum albumin. This was highly specific for 

galanthamine, and showed practically no cross reactivity against other common 

Amaryllidaceae alkaloids. However, other minor cross-reactive materials were 

present in a crude extract of Leucojum aestivum. This procedure was very sensitive 

(measuring range 0.5–100 ng), and was used to determine the galanthamine 

content of crude extracts of several L. aestivum samples, as well as of a number of 

South African Amaryllidaceae. 

The same group subsequently replaced the radio-labelled antigen with enzymelabelled 

antigen, thereby avoiding the use of radioactive material (Poulev et al., 

1993). This enzyme immunoassay procedure was easier to perform and more 

sensitive by a factor of 100, and required only a very small amount (1–12 mg) of 

plant material. By using this method, galanthamine contents of 1000 individual 

Leucojum aestivum plants and of more than 130 herbarium samples of Amaryllidaceae 

and closely related plant families were determined. Preliminary investigation 

showed that this method could be applied to analyse galanthamine in 

biological fluids, and the results generated by this method were in good agreement 

with those obtained by an HPLC method. This method was later used to 

determine the galanthamine content in bulbs and callus of two Pancratium species 

(Sarg et al., 1996). 

Quantitative determination of galanthamine by 

acetylcholinesterase inhibition 

Ghous and Townshend (1998) reported a method to determine galanthamine 

quantitatively by measuring its inhibition of acetylcholinesterase immobilised on 

controlled pore glass. The determination was carried out by a flow injection 

procedure where galanthamine and substrate, acetylthiocholine, were injected to 

coincide in the buffer stream, which then passed through the immobilised acetylcholinesterase 

column. Active enzyme cleaves the substrate to a chromogen 

reactive product. The eluent was mixed with a chromogen (5,5′-dithiobis-(2nitrobenzoic 

acid)) solution and the absorbance was measured at 405 nm. Under 

optimised conditions, the response was linear over the range 5 × 10 –7 to 6 × 10 –6 M. 

The limit of detection was 5 × 10 –7 M. Another group (Nikol’skaya et al., 1989; 

Kugusheva et al., 1992) also reported a procedure to determine galanthamine 

based on the same principle, using enzyme-containing membranes. 

Spectrophotometric and fluorometric determination 

Several simple spectrophotometric procedures have been developed to quantify 

galanthamine. These methods involved the measurement of absorbance in the UV 

range of galanthamine as a free base (λ 290 nm) (Bagdasarova, 1984) or in the 

visible range as a complex with other reagents (Kuznetsov et al., 1969; Pavlov and 

Ponomarev, 1981; Tokhtabaeva, 1987). Kolusheva and Vulkova (1966) studied the


UV spectra of galanthamine, lycorine and nivalidine. All three compounds had 

similar UV spectra with maxima at 222 and 286 nm. A quantitative spectrophotometric 

method was developed for the determination of galanthamine at 286 nm. 

The relationship between the fluorescence characteristics and chemical structure 

of galanthamine, lycorenine, lycorine, tazettine and vittatine was studied by 

Bruno and De Laurentis (1982). All these compounds had emission maxima at 

about 320 nm and excitation maxima at 290 nm. A linearity between the fluorescence 

intensity and concentration over 1–6 µM/l was observed, and the detection 

limit was 0.05 µg/ml. 

Burgudjiev and Grinberg (1993) analysed the UV and fluorescence spectra of 

galanthamine base and its hydrobromide in water, ethanol and n-hexane, and 

limits of detection for UV spectrophotometry and fluorometry were established. 

Yamboliev and Mikhailova (1985) described a spectrofluorometric method for 

the determination of galanthamine in biological fluids. Galanthamine was 

extracted using an organic solvent and re-extracted into a 0.1% aqueous sulphuric 

acid solution. The concentration of galanthamine was determined by spectrofluorometry 

using excitation and emission wavelengths of 286 and 315 nm, respectively. 

The fluorescence signal was linear in the range 0.05–15.0 µg/ml, and the detection 

limit was 0.05 µg/ml. However, due to insufficient specificity, the data on urine, 

bile and other pigmented fluids were unreliable. 

Several spectrophotometric and fluorimetric procedures have also been 

described to determine lycorine in Amaryllidaceae species. In these, lycorine was 

first separated by chromatographic methods and was quantitatively measured 

spectrophotometrically at 292 nm (Volodina et al., 1972, 1973; El-Din et al., 1983; 

Makhkmova and Safonova, 1994) or fluorimetrically at excitation and emission 

wavelengths of 292 and 330 nm, respectively (El-Din et al., 1983). 

Electrophoretic methods 

Gheorghiu et al. (1962) analysed the alkaloid extracts of Leucojum vernum and 

L. aestivum by a two-dimensional electrophoresis method. The analysis was carried 

out in alkaline medium and two compounds were detected and separated. 

Mikhno and Levitskaya (1971) described a paper electrophoresis method for the 

detection and quantification of galanthamine and securinine in biological material. 

The alkaloids were separated by paper electrophoresis at pH 2 and were detected 

with Dragendroff visualising agent. Galanthamine and securinine were eluted 

from the paper with 0.1N HCl and were quantitatively determined by spectrophotometry 

at 289 and 256 nm, respectively. 

Davey et al. (1998) reported a micellar electrokinetic capillary chromatographic 

method to analyse ascorbic acid and lycorine simultaneously in tissue extracts of 

Crinum asiaticum. Ascorbic acid and lycorine were extracted from the plant material 

using 3% metaphosphoric acid. Analysis utilised a 41 cm total length (34 cm to 

detector) × 50 µm fused silica capillary with a borate buffer (pH 9.0) and 50 mM 

sodium dodecyl sulphate as background electrolyte (applied voltage of +20 kV). 

Compounds were detected at 185 nm by fixed-wavelength detector or at 202 and 

267 nm by dual-wavelength detector. A linear detector response was observed for 

lycorine between 17 and 700 µM. The detection limits at 185 and 202 nm were 

17 and 1 µM, respectively.

Polarographic methods 


Occasionally, polarographic methods have been used to determine Amaryllidaceae 

alkaloids quantitatively. Volodina et al. (1970) determined the polarographic 

constants of galanthamine, lycorine, pancratine, hordenine and dl-nawardine 

isolated from Ungernia victoris, and used this procedure to quantify galanthamine 

isolated from a chloroform extract of this species by preparative TLC. In a 

subsequent study, Volodina et al. (1976) examined nine alkaloids isolated from 

Ungernia polarographically. They found that, within certain limits, there was a 

linear relation between the maximum current and the concentration that could be 

used in analytical studies. Recently, Meng et al. (1998) reported an oscillopolarographic 

titration method to determine galanthamine hydrobromide content in drug 

form, using silver nitrate as the titrant, with potassium nitrate and sulphosalicylic 

acid as supporting electrolytes. 

Counter current chromatography (CCC) 

Ma et al. (1994) described a pH-zone-refining CCC procedure for the separation 

of crinine, powelline and crinamidine from Crinum moorei using a multilayer coil 

planet centrifuge. Methyl tert-butyl ether containing triethylamine (5–10 mM) and 

water containing HCl (5–10 mM) were used as the solvent system. The separation 

was performed by using either the organic (displacement mode) or the aqueous 

phase (reverse-displacement mode) as the mobile phase. Compounds were eluted 

as an irregular rectangular peak and were separated into three plateaus by a UV 

detector or by pH measurement of the fractions. TLC analysis of the fractions 

showed a successful separation of the compounds with very narrow mixing zones. 

Results observed in both displacement and reverse-displacement modes were 

similar; however, the compounds were eluted in the reverse order. 

Gravimetric methods 

Amico et al. (1980) used a gravimetric method to quantify lycorine in various parts 

of Sternbergia lutea during the different stages of its life cycle. The crude alkaloid 

fraction obtained after traditional base neutral separation was crystallised from 

90% ethanol to yield lycorine in crystalline form. Lycorine was separated and 

its percentage based on dry weight was calculated. 

Variations in the formation of pancratistatin and related isocarbostyrils in 

Hymenocallis littoralis were studied by Pettit et al. (1995a) by a gravimetric method. 

Pancratistatin, narciclasine, 7-deoxynarciclasine and 7-deoxy-trans-dihydronarciclasine 

were isolated from H. littoralis bulbs by a series of extraction, precipitation 

and separation procedures, and the variation in their content during a period of 

one year was determined. 

CONCLUSIONS 

A number of methods have been developed during the last four decades to analyse 

Amaryllidaceae alkaloids quantitatively. In recent years, reversed-phase HPLC

Table 11.1 GC and HPLC procedures for the identification and quantitation of Amaryllidaceae alkaloids 

References 

Detector Detection 

limit 

Mobile phase 

(modifier) 

Method Test Compoundsa (Internal standard) Stationary phase or 

column 

SE-30 Argon FID Lloyd et al., 

1960 

PC-GC Galanthine, Acetylcaranine, Lycorenine, 

Galanthamine, Crinine, Powelline, 

Tazettine, Belladine 

PC-GC Galanthamine, Lycoramine, Tazettine SE-30 Yamaguchi 

et al., 1962 

PC-GC Norpluviine, Lycorine, Lycoramine, SE-30, XF-1105, Nitrogen FID Takagi 

Galanthamine, Buphanamine, Vitattine, XE-60, XF-1150, 

et al., 1968a 

Crinine, Tazettine, Haemanthamine, ECNSS-S, NGS, 

Crinamidine, Hippeastrine, Undulatine, PEG-20M, EGSS-X, 

Homolycorine (Chrysene) 

HI-EFF 8B 

PC-GC Lycoramine, Galanthamine HI-EFF 8B Nitrogen FID Takagi 

et al., 1968b 

PC-GC-MS Ambelline, Criwelline, Crinamine, OV1 Helium Ion current 

Millington 

Lycorine, Criglaucine, Criglaucidine 

(MS) 

et al., 1972 

PC-GC-MS Lycorine, Ornazidine, Ornazamine, OV-17 Ion current 

Onyiriuka 

Crinatine 

(MS) 

et al., 1978 

CGC-MS Galanthamine, Lycoramine, 

DB-1, DB-5 Helium Ion current 

Kreh et al., 

N-Demethyl-galanthamine, 

(MS) 

1995a 

epi-Norlycoramine, O-Methyloduline, 

Vittatine, Narwedine, Demethylpluviine, 

10-O-Demethylpluviine, Oduline, 

Lycorenine, Haemanthamine, Masonine, 

Homolycorine, N-Demethyl-masonine

DB-1 Helium FID/NPD Bastos 

et al., 1996 

CGC Galanthamine, Lycoramine, 

N-Demethyl-lycoramine, 

O-Methylmaritidine, Caranine, 

Pluviine, Haemanthamine, Lycorine, 

Homolycorine, Hippearstrine, 

Narcissidine (Deoxydihydrocodeine) 

HPLC Ambelline C-8 Methanol, Water Circular 

Westwood 

(Ammonia) 

dichroism 

et al., 1981 

(CD) 

HPLC Lycorine Perkin-Elmer Acetonitrile, Aqueous UV (290 nm) 5 ng Evidente 

C18/10 

ammonium carbonate 

et al., 1983 

HPLC Lycorine, Tazettine LiChrosorb-CN Methanol, Water Fluorescence


References 

Detector Detection 

limit 

Mobile phase 

(modifier) 

Method Test Compoundsa (Internal standard) Stationary phase or 

column 

Eckers 

et al., 1976 

5 ng/ml Claessens 

et al., 1983 

Ion current 

(MS) 

UV 

(235 nm) 

10 ng/ml Claessens 

et al., 1988 

0.05 µg/ml Tencheva 

et al., 1987a 

UV 

(235 nm) 

UV 

(280 nm) 

1 ng/200 µl Bickel et al., 

1991 

Fluorescence 

(excitation 

290/emission 

320 nm) 

LC/MS Lycorine, Ambelline, Criglaucine ODS 5 µ Spheresorb Water, Acetonitrile, 

Ammonia 

HPLC Galanthamine (Phenacetin) CP tm Micro Spher Si n-Hexane-Dichloro- 

5 µm 

methane-Ethanolamine 

HPLC Galanthamine Novapack C-18 (4 µ) Acetonitrile-Pentanesulfonic 

acid buffer 

HPLC Galanthamine, Epigalanthamine, Hibar-LiChrosorb Methanol-Water 

Galanthaminone (Codeine) 

RP-8 (5 µm) (Dibutylamine, 85% 

Phosphoric acid) 

HPLC Galanthamine, Epiganthamine 

Intersil-C-8 silica Acetonitrile, Tetra- 

(Codeine) 

(5 µm) 

hydrofuran, Water 

(di-n-Butylamine, 

85% Phosphoric acid) 

Note 

a 

Reference compounds or those detected.


and CGC have emerged as the most accurate and practical analytical procedures 

to determine these compounds in natural sources and in biological samples. 

HPLC was the most widely used technique for this purpose, especially when only 

one constituent was quantified. Due to the presence of at least one aromatic ring in 

their chemical structures, Amaryllidaceae alkaloids can be conveniently quantified 

with high sensitivity by a UV or a fluorescence detector. This method is not always 

suitable for the routine analysis of Amaryllidaceae alkaloids in natural sources. 

Usually, alkaloid fractions of Amaryllidaceae contain complex mixtures of compounds 

with diverse chemical structures and polarities. It is often difficult to 

achieve good resolution of these mixtures with reasonable peak shapes, even with 

complex solvent programming. The resolution and peak shapes for a mixture of 

standards also appear to vary widely under identical experimental conditions for 

commercially available reversed-phase columns with similar specifications from 

different suppliers. 

CGC appears to be a better technique for the simultaneous quantification of 

several different alkaloids in natural sources. Amaryllidaceae alkaloids can be 

analysed by this method without any prior derivatisation. However, the method is 

not applicable to quaternary alkaloids and alkaloid N-oxides. Compounds usually 

show very sharp peaks and can be detected using a flame ionisation detector 

(FID), a nitrogen phosphorus detector (NPD) or mass spectrometry (MS). A very 

high resolution, even for complex mixtures of alkaloids, can be achieved with 

relatively short capillary columns within a reasonable experimental time. Two of 

the most common Amaryllidaceae alkaloids, haemanthamine and lycorine, were 

reported to undergo partial decomposition under GC experimental conditions. 

These compounds still appear as relatively sharp single peaks, with good concentration/response 

linearity in a wide range of concentrations, and can be quantified 

reproducibly. These analyses can be performed with little sample preparation, 

and are suitable for routine quantitative analysis of Amaryllidaceae alkaloids in 

natural sources. Useful information on these two analytical procedures is summarised 

in Table 11.1. 


We express our gratitude to Dr. Larry Walker for his many suggestions and for 

reviewing the manuscript. 

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12 Synthesis of galanthamine and 

related compounds 

V.N. Bulavka and O.N. Tolkachev 

INTRODUCTION 

(–)-Galanthamine is the principal representative alkaloid from the Amaryllidaceae 

family, possessing a characteristic benzofuro[3a,3,2-ef ][2]benzazepine skeleton. 

It is an anticholinesterase agent of low toxicity used in medicine for the treatment 

of a number of conditions (Mashkovsky and Kruglova-L’vova, 1951; Mashkovsky, 

1955). It is under evaluation for the treatment of Alzheimer’s disease (Fulton and 

Benfield, 1996; Moraes-Cerdeira et al., 1997). 

Galanthamine hydrobromide was produced commercially as ‘Nivalin’ in Bulgaria 

from Galanthus nivalis L. and later from Leucojum aestivum cultivars, and in the 

former USSR from leaves of Ungernia victoris L. collected in the mid-Asia regions. 

The limited plant sources available in the Russian flora for commercial exploitation, 

the low concentration of the alkaloid in plant material and the labourconsuming 

character of the chemical processes, led to a need for alternative routes 

of galanthamine production. At present, synthetic methods seem to have assumed 

practical significance. 

The chemistry of Amaryllidaceae alkaloids has been extensively reviewed 

(Abduazimov, 1993; Cook and Loudon, 1952; Fuganti, 1975; Grundon, 1984, 

1985, 1987; Jeffs, 1990; Lewis, 1990, 1997; Polt, 1996). Today about 20 natural 

alkaloids of the galanthamine type, isolated from species of Amaryllidaceae, are 

known. Up to now, five alkaloids – galanthamine, narwedine, lycoramine, sanguinine 

and N-norgalanthamine – have been obtained by total synthesis. 

CHEMICAL PROPERTIES OF GALANTHAMINE 

(–)-Galanthamine ((–)-1) is a strong base, which on interaction with alkylhalogenides 

produces quaternary ammonium salts. The oxidation of (–)-1 with MnO 2 

in mild condition leads to the ketone 2, isolated in optically inactive form (Barton 

and Kirby, 1962; Combes and Lefebvre, 1962). Its (–)-enantiomer is the natural 

alkaloid known as narwedine (Figure 12.1). Narwedine readily undergoes racemisation 

in alcoholic solutions, especially in the presence of organic bases and acids, 

or spontaneously without any additives (Barton and Kirby, 1960, 1962). The 

mechanism of this process has been explained by the formation of tautomeric 

dihydrofuranoic and hydroxyspirodienone (3) forms, which are converted in a

MeO 

MeN 

HO 

O 

HO 

(–)-1 

OH 

O 

NMe 

OMe 

NR 

5 (R=H) 

6 (R=Me) 

OH 

MeO 

MeO 

MeN 

OR 

O 

HO 

O 

O 

Synthesis of galanthamine 305 

equilibrium process as shown in Figure 12.1. Enantiomeric (–)-2, on reduction 

with different agents, produces a mixture of (–)-1 and (–)-epi-galanthamine ((–)-4). 

The key compound in the biosynthesis of (–)-1 and related Amaryllidaceae 

alkaloids is norbelladine (5, R = H; Figure 12.1), which is formed from tyrosine 

(Barton and Cohen, 1957; Barton et al., 1961, 1962, 1963; Schutte, 1969). 

O 

3 

O 

NMe 

NMe 

OMe 

MeO 

MeN 

O 

HO 

OH 

(–)-2 (–)-4 

(+)-1 (+)-2 (+)-4 

Figure 12.1 (Above) The equilibrium process between narwedine hydrohydienone (3) and 

its enantiomers (2) and oxidative/reductive interconversion between (2) and 

galanthamine (1) and epi-galanthamine (4) enantiomers. (Below) Norbelladine 

(5) and N,O-dimethylnorbelladine (6). 

O 

NMe 

OMe

306 V.N. Bulavka and O.N. Tolkachev 

THE SYNTHESIS OF ALKALOIDS OF THE GALANTHAMINE GROUP 

The first synthesis confirming the structure of galanthamine 

Galanthamine was isolated for the first time from Galanthus woronowii and its 

preliminary structure was proposed from data on the chemical degradation of its 

molecule using classical methods (Proskurnina and Yakovleva, 1952, 1955). The 

final confirmation of the structures of galanthamine, narwedine and lycoramine 

were made by their total synthesis. The first biomimetic route of (±)-2, and after its 

resolution, of (+)-1, (–)-1 and (–)-lycoramine synthesis, proposed by Barton and 

Kirby (1960, 1962), included the oxidation of diphenolic substrates such as O,Ndimethyl-norbelladine 

(6, R = Me), the biogenetic precursor of 1, or its derivatives 

(Figure 12.2). The synthesis was carried out starting from 4-hydroxybenzaldehyde 

(7) through cyanohydrine (8) and 4-hydroxyphenylacetic acid (9) (34%), which, 

after O-benzylation into 10 (67%), was converted into the acid chloride 11 (Figure 

12.2). The coupling component was obtained from 3-benzyloxy-4-methoxybenzaldehyde 

(12) via the intermediate Schiff base (13), with a subsequent reduction 

to 3-benzyloxy-4-methoxy-N-methyl-benzylamine (14) (68% yield). With the 

acid chloride 11, the latter yielded the corresponding arylacetamide 15 (85%). The 

LiAlH4 reduction of 15 (88%) and Pd/C hydrogenolysis of the intermediate 

tertiary amine 16 produced the desired compound 6 in 76% yield. Oxidation with 

different agents produced the target ketone (±)-2 in very low yield (0.1–1.4%). The 

best results were obtained on K3 [Fe(CN) 6 ] oxidation (0.92–1.4%). The final LiAlH4 reduction afforded a mixture of (±)-1 and (±)-4 in the ratio 3:2 (ca. 100%), from 

which the first compound was isolated in crystalline form (39%). Aluminium 

isopropoxide reduction produced preferentially the epi-isomer (±)-4 (60%). The 

total yield of (±)-2 was 0.18%, and of (±)-1, only 0.07%. 

Also proposed was a method of conversion of the ketone (±)-2 into enantiomeric 

(–)-1 via a step of dynamic resolution of (±)-2 to (+)-2 in the presence of optically 

active additives (–)-1, (–)-4 or (–)-lycoramine, and the LiAlH4 reduction of (+)-2 to 

yield a mixture of (+)-1 and (+)-4. The latter were used for seeding during the 

conversion of the racemate to (–)-2. A similar LiAlH4 reduction of (–)-2 produced 

the target (–)-1 in 58% yield. The subsequent hydrogenation over Pd/C afforded 

(–)-lycoramine (Barton and Kirby, 1960, 1962). Recently, a modified version of the 

pilot plant method of dynamic resolution of (±)-narwedine has been applied 

(Shieh and Carlson, 1994; Czollner et al., 1998). 

Synthesis of narwedine via palladium-directed coupling 

The synthesis was modified by Holton et al. (1988) as follows (Figure 12.3). 

A hydroxy group in 3-hydroxy-4-methoxybenzaldehyde (18) was protected by the 

action of NaH in OP(NMe 2 ) 3 , N 2 , then MeSCH 2 Cl; the intermediate O-methylthiomethyl 

derivative (19) (Holton and Davis, 1977), on condensation with 4-hydroxyphenethylamine 

(17), gave the Schiff base 20, which, after NaBH 4 reduction to 

21, hydroxymethylation and NaB(CN)H 3 reduction of 22, gave the N-methylderivative 

23 in 90% overall yield. With LiPdCl 4 in MeOH-(i-Pr) 2 NH (–78 °C), 23 

produced the palladium derivative (±)-24 as a diastereomeric mixture (95%). 

Oxidation with (CF 3 COO) 3 Tl (–10 °C) gave the expected compound (±)-25, which

OH 

O 

MeO 

PhCH 2O 

CHO 

O 

PhCH 2O 

OH 

HO N 

OCH 2Ph 

NMe 

OMe 

PhCH 2O 

PhCH 2O 


on hydrolysis produced (±)-26 (98%). (±)-25 on keeping at 25 °C in the presence of 

Ph 3 P (14 h) yielded (±)-2 (51%). Thus the total yield of (±)-2 was 43.6%. 

Hence, the concept of using metal-substituted substrate-induced o-diphenol oxidation 

in galanthamine synthesis was successful. However, the use of toxic, expensive 

and scarce compounds means that the scheme is unsuitable for large-scale application. 

Synthesis of galanthamine and N-norgalanthamine 

via belladine-derived amide oxidation 

OH 

Me 

N 

MeO OCH 2Ph 

A significant achievement in the synthesis of 1 was found in the work of Kametani 

et al., 1969a,b and Kametani, 1972a, shown on Figure 12.4. 5-Benzyloxy-2-bromo- 

NMe 

OCH 2Ph 

7 8 9 10 11 

OH 

OMe 

OH 

MeO OCH 2Ph 

12 13 14 

O 

O 

HO 

HO 

OCH 2Ph 

Cl 

Me 

NH 

NMe 

15 16 6 

O 

OMe 

Figure 12.2 Synthesis of narwedine (2) via N,O-dimethylnorbelladine (6) (Barton and 

Kirby, 1960, 1962). 

2


MeO 

CHO 

17 

CHO 

O OMe O OMe 

MeS 

MeS 

OH MeO O 

SMe 

18 19 20 21 

MeS 

Me 

HO 

NH 2 

HO HO 

O 

O + 

O 

N OH 

O 

OMe 

MeS 

2 

O 

25 26 

4-methoxy-benzaldehyde (29) was oxidised with silver oxide into the corresponding 

acid 30 (89%), which was then transformed into the acid chloride 31 (52%). 

The coupling component 28 was obtained from 4-benzyloxyphenylacetic acid (10) 

via the acid chloride 11, N-methyl acetamide (27) (91%) and final reduction to 28 

NMe 

HO 

OMe 

N 

Me 

N 

OMe 

22 23 24 

NMe 

PdCl 

SMe 

O 

O 

HO 

Cl 

Pd 

SMe 

O 

O 

OH 

NH 

NMe 

PdCl 

SMe 

Figure 12.3 Synthesis of narwedine (2) via palladium-containing intermediate (24) 

(Holton et al., 1988).

10 

Br 

11 

CHO 

MeO OCH 2Ph 

Br 

27 28 

CO 2H 

MeO OCH 2Ph 


COCl 

with LiAlH 4 in 82% yield. Coupling 31 with 28 produced the corresponding 

benzamide 32, which on acidic hydrolysis afforded the 4-hydroxy compound 33 in 

63% yield. The K 3 Fe(CN) 6 oxidation of 33 produced the key compound (±)-34 

Br 

MeO OCH 2Ph 

29 30 31 32 

MeN 

Br O 

MeO 

OH 

OH 

MeO 

OCH 2Ph 

HO 

MeO 

33 34 

O 

NHMe 

O 

35 

O 

NMe 

O 

OCH 2Ph 

NHMe 

O 

Br 

NMe 

O 

MeN 

Br 

MeO 

1+4 

O 

OCH 2Ph 

OCH 2Ph 

Figure 12.4 Synthesis of galanthamine (1) via belladine-type benzamide (33) (Kametani 

et al., 1969a,b; Kametani, 1972a).


(40%), together with the debrominated spiro-dienone 35 (0.4%). Ketone (±)-34, on 

LiAlH4 reduction, produced a mixture of (±)-1 (50%) and (±)-4 (40%), which were 

separated chromatographically. Thus, the total yield in the synthesis of (±)-1 as 

shown in Figure 12.4 was 9.4%. The use of the amide synthone 33 was successful in 

the oxidation reaction. The yield from cyclocondensation at this step was 29 times 

higher than in case of tertiary amine oxidation used in Barton and Kirby’s scheme. 

However, taking into consideration that from the starting material 10 (not a com- 

7 

CHO 

OCH Ph 

2 

CHO 

N 

36 10 

MeO OMe 

MeO OH 

MeO 

Br 

37 

CN 

+7 

CO H 

2 

38 18 12 

MeO 

OCH Ph 

2 

OCH Ph 

2 

OCH Ph 

2 

CHO CHO 

CHO 

Br 

OH MeO 

39 29 

OCH Ph 

2 

CHO 

OCH Ph 

2 

Figure 12.5 Synthesis of 4-benzyloxyphenylacetic acid (10) (above; Vaghani and Merchant, 

1961) and of 2-bromo-4-methoxy-5-benzyloxy-benzaldehyde (29) 

(below; see references in text).


mercially available compound) these authors obtained a 23% yield in three stages, 

the yield of (±)-1 from 7 (which is commercially available) is 2.14% (30 times higher). 

Kametani et al. (1969a,b) used the following route for the synthesis of 4-benzyloxyphenylacetic 

acid (10) (Figure 12.5). 10 was obtained from 7 through the 

aldehyde 36 and the intermediate imino-propionitrile of structure 37 (Vaghani and 

Merchant, 1961). However, in this scheme, the total yield of 10 was not indicated, 

and only half of the starting material was inserted into the target compound. 

Another component, 2-bromo-4-methoxy-5-benzyloxy-benzaldehyde (29), was 

obtained in three steps starting from 3,4-dimethoxybenzaldehyde (38) by 3-monodemethylation 

with concentrated H 2 SO 4 to isovanilline (18) (61%) (Brossi et al., 

1967), and bromination with bromine in acetic acid to give the expected compound 

39 in 67% yield (Henry and Sharp, 1930; Raiford and Ravely, 1940). The 

total yield of 29 after O-benzylation was about 33% (Jackson and Martin, 1966), 

which after total conversion should exceed 54%. Recently, a more advanced route 

(Figure 12.5) was proposed from isovanilline (18), which after O-benzylation to 12 

(98%) and bromination in acetic acid in the presence of NaOAc produced 29 in 

79% yield (Bolton et al., 1987), or a total 47% yield (77% taking into consideration 

a complete conversion of products on the hydrolysis reaction step). 

Similarly, Kametani et al. (1973) synthesised (±)-1 via (±)-N-norgalanthamine 

((±)-44) (Figure 12.6). The substrate for oxidation was obtained in three steps from 

the acid 30 via the acylchloride 31, coupled with the amine 40 to produce the amide 

41, which was debenzylated to 42 in 72% total yield. On K 3 [Fe(CN) 6 ] oxidation, the 

key compound (±)-43 was obtained in only 0.7% yield. The latter was reduced with 

LiAlH 4 to (±)-44 in 32% yield. The final formaldehyde-formic acid methylation of 

(±)-44 produced the desired (±)-1 in 70% yield, corresponding to 0.11% total yield. 

In another synthesis of (±)-1, Kametani et al. (1971a) used the intermediate 

isomeric amide 48 (a positional carbonyl isomer of 33). Starting from 4-benzyloxyphenylacetic 

acid (10) converted to acyl chloride (11), and the bromoamine 46 

obtained from bromoaldehyde (29) in two steps without isolation of the intermediate 

45 (75%), the desired amide 47 was produced in 72.3% yield (Figure 12.6). 

After elimination of protective benzyl groups with hydrobromic acid, the resulting 

dihydroxy-amide 48 was oxidised with K 3 [Fe(CN) 6 ] to afford a tetracyclic amide of 

structure (±)-49 in low yield (1.9%). A subsequent LiAlH 4 reduction afforded 

12.3% of (±)-1 and 4.9% of (±)-4. Thus, the total yield of the title compound 

according to this scheme was 0.095%, which was 99 times lower than that according 

to the alternative route via the intermediate 33. 

The oxidation of unprotected amide 52 (obtained from 28 and 50 via 51) with 

VOCl 3 produced the expected cyclic compound (±)-53 in 2% yield (Figure 12.7), 

which was reduced to (±)-1 and (±)-4, with (±)-1 isolated in 61% yield. The 0.77% 

total yield of the latter is 12 times lower than that shown according to the route in 

Figure 12.4 with the application of bromo amide (Kametani et al., 1971b; Kametani, 

1972b). 

In another modification of (±)-1 synthesis, Kametani et al. (1972) used the 

isomeric 2-bromo derivative 54 as a starting material (Figure 12.8). The latter was 

transformed via 55 (63%) and 56 (86%) into the acid chloride 57, and coupled with 

the methylamine derivative 28 to give the amide 58 (37%). Hydrochloric acid 

debenzylation produced diphenol 59 (77%), which on irradiation with a mercury 

lamp afforded the key compound (±)-53 in only 1% yield. LiAlH 4 reduction as


PhCH O 

2 

MeN 

Br 

MeO 

MeO 

NH2 

O 

O 

30 

31 

O 

NH 

O 

NH 

MeO Br 

MeO 

O 

Br 

43 44 

OH 

47 48 49 

O 

O 

HO 

O 

HO 

HO 

O 

O 

NH 

MeO Br 

40 41 42 

29 

OCH Ph 

2 

OCH Ph 

2 

PhCH O 

2 

PhCH O 

2 

Br N 

Me 

NH 

Me 

NH 

MeO OCH Ph MeO OCH Ph 

Br 

45 46 

MeN 

Br 

MeO 

2 

OH 

MeO 

Br 

2 

NMe 

O 

10 

11 

1 

1 + 4 

Figure 12.6 Synthesis of galanthamine (1) via N-norgalanthamine (44) (above; 

Kametani et al., 1973) and via belladine-type phenylacetamide (48) 

(below; Kametani et al., 1971a).

MeO 

COCI 

OCH Ph 

2 

28 

50 51 52 

MeO 

O 

53 

O 

NMe 

O 

MeN 


above produced a mixture of (±)-1 and (±)-4. A total yield of 0.093% was obtained 

using this scheme, which was 40 times lower than in the case of (±)-1 synthesis via 

isomeric bromoamide. Thus Kametani’s group demonstrated the necessity of 

blocking the para-position to the hydroxy group by bromination in the benzyl 

moiety in the phenolic coupling process. 

Subsequently, Bulgarian and German scientists tried to improve the synthesis of 1. 

Vlahov et al. (1978), in a modified route of the synthesis, used 4-methoxybenzylation 

for hydroxyl group protection and subsequent de-protection in mild conditions. 

Thus, intermediate amides 60 and 61 were transformed to 33 and 48, 

respectively (Figure 12.9). However, the final cyclocondensation of 48 to 49 (as in 

figure 12.6) proceeded with low yields. In another variation of the synthesis by the 

same authors, in order to avoid the use of LiAlH 4 reduction of the amide 62, the 

following succession was applied: a transformation of 62 to thioamide 63 by P 2 S 5 - 

pyridine treatment (58% yield) with subsequent NaBH 4 -CoCl 2 reduction to the 

substituted N-methyl-phenethylamine (64) in 80% yield. The oxidative cyclocondensation 

of the amide 33 produced the desired racemic bromo-ketone (±)-34 in a 

O 

OCH Ph 

2 

MeO OCH2Ph MeO 

1 + 4 

Figure 12.7 Synthesis of galanthamine (1) via non-brominated belladine-type benzamide 

(52) (Kametani, 1972b; Kametani et al., 1971b). 

MeN 

OH 

O 

OH


CHO 

Br 

MeO OH MeO OCH Ph 

54 55 56 

MeO 

COCI 

Br 

OCH Ph 

2 

28 

57 58 59 

1 + 4 

CHO 

MeN 

Br 

2 

O 

Br 

MeO 

OCH Ph 

MeO 

2 

OCH Ph 

2 

MeO OCH Ph 

2 

yield after recrystallisation below 15% (Vlahov et al., 1978, 1989). The Bulgarian 

group also used microbiological reduction of (–)-34 to (–)-bromogalanthaminone 

((–)-65) with Nematospora coryli and to (–)-bromo-epi-galanthamine ((–)-66) with 

Septomyxa affinis (Vlahov et al., 1984) (Figure 12.9). 

Synthesis of enantiomeric galanthamine starting from chiral synthone 

Shimizu et al. (1977, 1978) (Figure 12.10) have synthesised (+)-1 starting from its 

natural precursor L-(+)-tyrosine methyl ester (67), which, after condensation with 

3,5-dibenzyloxy-4-methoxybenzaldehyde (68) and NaBH 4 reduction, without the 

isolation of the corresponding Schiff base, was converted into L-(+)-N-(3,5-dibenzyloxy-4-methoxybenzyl)tyrosine 

methyl ester (L-(+)-69). Its trifluoroacetyl derivative 

L-(–)-70 was debenzylated by Pd/C hydrogenolysis (100%) and the trihydroxy 

compound L-(–)-71 was oxidised with Mn(III) acetylacetonate in acetonitrile into 

the tetracyclic (+)-72 (34%); its (–)-stereomer (–)-72 was not isolated. The hydroxy 

group was phosphorylated with diethylphosphoryl chloride to produce (+)-73 

and its (–)-stereomer ((–)-73) (13:1), with the former isolated in 81% yield. NaBH 4 

reduction followed, with trifluoroacetyl group elimination giving a mixture of the 

epimeric alcohols (+)-74 (axial) and (+)-74 (equatorial) (4:1). The key compound 

(+)-74 (axial) was isolated in 21% yield on formaldehyde-formic acid methylation, 

MeN 

CO H 

2 

Figure 12.8 Synthesis of galanthamine (1) via photochemical substitution of bromine in 

belladine-type benzamide (59) (Kametani et al., 1972). 

OH 

O 

Br 

Br 

OH 

53

MeO 

MeO 

MeO 

HN 

Me 

O 

O 

O 

O 

OMe 

O 

Br 

Me 

N 

HN 

Me 

S 

O 


HN 

Me 

and subsequent aminolysis produced (+)-carboxamide ((+)-75) (37%). The O-acetyl 

derivative 76 (89%) was transformed into the cyano derivative 77 (POCl 3 ), and 

LiAlH 4 reduction afforded benzazepine of the structure (+)-78 in 42% yield. The 

OMe 

OMe 

O 

OMe 

62 63 64 

HO 

(–)-65 

Br 

O 

NMe 

O 

MeO 

Me Br 

N 

O 

60 

61 

O 

OMe 

O 

OMe 

O 

OMe 

OH 

O 

O 

NMe 

MeO 

O 

Br 

Br 

(–)-34 (–)-66 

Figure 12.9 (Above) 4-Methoxybenzyl-protected intermediates for the synthesis of galanthamine 

(1) (Vlahov et al., 1978, 1989). (Below) Microbiological reduction of 

bromo-narwedinone (34) (Vlahov et al., 1984). 

NMe


MeO 

O 

PhCH2O 

67 

MeO OCH2Ph 

68 69 70 

71 

R=H R=COCF3 

MeO 

O 

F3C 

H2NCO 

MeN 

NH2 

O 

N 

OH 

CHO 

O 

O 

O 

MeO 

O 

O 

MeO 

O 

O 

OMe 

O 

N 

OMe 

HN 

OMe 

72 

OH 

F3C 

73 

OPO(OEt)2 

OPO(OEt)2 

74 

OR 

HO 

MeOCO 

NR 

PhCH2O OMe 

O 

NC 

OMe 

MeN 

OPO(OEt)2 

O 

OCH2Ph 

Me 

O 

final protection group elimination with Na-liquid NH 3 gave the expected compound 

(+)-1 in 72% yield (0.63% total yield). 

Despite the low total yield of galanthamine, this work was of a great importance 

because of its prospects for further improvement. Thus, the tyrosine hydroxyl 

group was not protected, and this shows that the protection and de-protection of 

O 

OMe 

OPO(OEt)2 

MeN 

75 76 77 78 

R=H R=COCH3 

HO 

MeOCO 

N 

COCF3 

HO OMe 

OH 

OH 

OH 

O 

OMe 

OPO(OEt)2 

Figure 12.10 Synthesis of enantiomeric galanthamine (1) (Shimizu et al., 1977, 1978). 

(+)-1

18 

17 

HO 

HO 

HO 

N 

OMe 

HO 


the phenolic hydroxyl groups was not necessary. The amide modification of the 

N-methyl part of the substrate, and the successful oxidation in homogeneous 

conditions, were also important methodological findings. 

Galanthamine synthesis via N-formylnorbelladine derivative 

HO 

NH 

The logical continuation of the syntheses shown in Figures 12.5 to 12.10 was realised 

in the improved method of galanthamine synthesis proposed by Szewczyk et al. 

(1988, 1995) (Figure 12.11). This involved a six-step scheme starting from 4-hydroxyphenethylamine 

(17) and isovanilline (18) via the Schiff base 79, which was 

reduced without isolation using NaBH 4 to a secondary amine 80 in 99% yield. The 

latter, on treatment with ethyl formate, gave the corresponding formamide 81 in 

80% yield, which on bromination with bromine in chloroform – methanol mixture 

(–65 °C) afforded the bromo-formamide 82 (85%). K 3 [Fe(CN) 6 ] oxidation gave the 

expected benzazepine 83 (21%). The final LiAlH 4 reduction produced a mixture 

OMe 

HO 

HO 

79 80 81 

N 

CHO 

Br OH 

OMe 

MeO 

O 

Br 

82 83 

O 

N 

N 

CHO 

CHO 

OMe 

1+4 

Figure 12.11 Synthesis of galanthamine (1) via N-formylnorbelladine derivative (82) 

(Szewcyzk et al., 1988, 1995).


of 53% (±)-1 and 31% (±)-4. The total yield of (±)-1 according to this scheme was 

7.5%, which exceeded by 3.5 times the results published in the work of Kametani 

et al. (1969a,b). Taking into account that isovanilline (18) is usually obtained from 

veratric aldehyde (38) in 61% yield (100% conversion) (Brossi et al., 1967), in this 

scheme Szewczyk et al. (1988, 1995) used all commercially available compounds. 

The synthesis seems to be simple in realisation on a preparative scale, but its basic 

deficiency is the use of the inflammable LiAlH4 . 

Galanthamine synthesis developed by the Russian group 

A comparative analysis of the known approaches to galanthamine synthesis has 

revealed the best route to the production of the key compound containing the 

carbonyl moiety as the formamido group, shown in Figure 12.11. The failing of 

the majority of known methods is the low yields obtained. The present authors 

have used the following strategy in a large-scale approach to its synthesis: the 

use of cheap and commercially available materials, a minimum number of steps, 

and the use of synthones with unsubstituted hydroxy groups. We have proposed 

the synthesis of (±)-1 from 4-hydroxyphenethylamine (17), or its O-alkyl derivatives, 

and 2-bromo-5-hydroxy-4-methoxy-benzaldehyde (39) (Bulavka, 1993; 

Bulavka et al., 1990, 1991, 1993, 1994a,b,c, 1999; Bulavka and Tolkachev, 1995) 

(Figure 12.12). 

The known methods of parent compound production were modified. Thus, 

4-hydroxy- and 4-alkoxyphenethylamines (86, R = H, Me, Et, Pr, PhCH 2 ) were 

obtained from the corresponding 4-substituted benzaldehydes (84) via substituted 

2′-nitrostyrenes (85) through zinc dust reduction in diluted hydrochloric acid or 

in a mixture of hydrochloric and acetic acids. The method was optimised to produce 

the expected 4-hydroxy- and 4-alkoxyphenethylamines in 80–98% yields in 

one operation (Bulavka et al., 1993, 1994a,c), higher yields than in earlier published 

methods. Primary amines 86 afforded formamides 87 (90–95%) with formic 

acid. 87 (R = Me) was reduced to N-methylated amine 91 with NaBH 4 – acetic acid 

(74%) and alternatively with zinc dust and sulphuric acid in tetrahydrofouran 

(67%) (Bulavka, 1993; Bulavka et al., 1994c). The alternative method of N-methyl- 

4-methoxyphenethylamine and N-methyl-4-hydroxyphenethylamine production 

starting from 4-methoxyacetophenone (88) via N-methyl-4-methoxyphenylthioacetamide 

(89) was also studied. The Willgerodt-Kindler reaction of 88 and 

MeNH 2 (160–170 °C in ampoules) gave a complex mixture of products, from 

which 89 (10.4%) was isolated together with the by-product N-methyl-4-methoxyphenylacetamide 

(21.9%). The method was modified as follows: interaction of 

4-methoxyacetophenone (88) with methylamine in the presence of dehydrating 

agents gave the expected Schiff base 90 in 77% yield, which was then transformed 

to desired thioamide 89 in 21.5% yield. The reaction of 88 with methylamine 

hydrochloride and sulphur in dimethylformamide (100 °C, sodium acetate, or 60– 

70 °C, 4-toluenesulfonic acid – Et 3 N) gave 89 in 57–58% yields. The reaction of 88 

with MeNHCHO and sulphur (170–180 °C) afforded the expected 89 in 30–35% 

yield. Subsequent reduction of 89 with zinc dust and hydrochloric acid afforded 

N-methyl-4-methoxyphenethylamine (91) (91%), which was demethylated with 

hydrochloric acid at 170 °C to N-methyl-4-hydroxyphenethylamine (92) (98%) 

(Bulavka, 1993; Bulavka et al., 1999).

RO 

O 

RO 

R=H, Me, Et, Pr, PhCH2 

MeO 

38 

Br CHO 

MeO OMe 

NO2 

39 


Owing to the low yields of 89, preference was given to the route of (±)-1 synthesis 

via the intermediate Schiff base from 17 and 39. The latter was obtained 

from 3,4-dimethoxy-benzaldehyde (38) which was brominated regioselectively 

with bromine in dichloromethane to produce 2-bromo-4,5-dimethoxy-benzaldehyde 

(93) in 98% yield. The latter was selectively O-demethylated with sulphuric 

acid to afford 2-bromo-5-hydroxy-4-methoxy-benzaldehyde (39). The last reaction 

RO 

84 85 86 

O 

Me MeO 

88 89 

93 

N 

Me Me 

90 

MeO 

17 

82 83 1+4 

S 

HN Me 

HO 

MeO 

NH2 

NH 

Me 

N 

Br 

HO OMe 

HO 

RO 

HO 

91 92 

NH 

NH 

Me 

Br 

HO OMe 

94 95 

87 

NHCHO 

Figure 12.12 Synthesis of phenylethylamine intermediates (above; Bulavka, 1993; 

Bulavka et al., 1994a,c, 1999) and galanthamine (1) (below; Bulavka, 

1993; Bulavka et al., 1990, 1994b; Bulavka and Tolkachev, 1995).


was optimised to produce the key compound in 80% yield (92% conversion) 

(Bulavka et al., 1990). Isomeric 2-bromo-3-hydroxy-4-methoxy-benzaldehyde (54), 

suitable for photochemical cyclisation (as on Figure 12.8), was also obtained by 

highly regioselective bromination of isovanilline with N-bromosuccinimide (Bulavka 

et al., 1991). 

Synthesis of 2-bromo-O-methylnorbelladine (95) was carried out according to 

the scheme shown on Figure 12.12 via the Schiff base 94, preferably without its isolation. 

The reductive amination of 39 with 17 (molecular sieves 4 Å, then NaBH4 in 

MeOH) yielded the secondary amine 95 (85–96%), which after N-formylation 

(89%, or 99% conversion) to 82 (Bulavka et al., 1994b) was oxidised with Mn(III) 

acetylacetonate in acetonitrile to 83 (25%) (Bulavka and Tolkachev, 1995), and 

then converted to (±)-1 and (±)-2 (53% and 31%). The total yield of (±)-1 was 9.5%, 

higher than in the previous scheme. 

Industrial synthesis of galanthamine – recent modifications 

Czollner et al. (1997, 1998) recently modified the method of oxidation, optimising 

the phenol coupling process to 45–50%. The final product was obtained as the 

optically active compound. Phenolic oxidation of the formyl-tyramine derivative 

82 in a two-phase liquid system was proposed, while (±)-8-bromo-N-formylnornarwedine 

(83) was obtained in 26% yield (Henshilwood and Johnson, 1996). 

A similar route was described in a patent of Tiffin et al. (1997). Compound 83 was 

asymmetrically reduced to enantio-enriched (–)-galanthamine in 36% yield and 

50% enantiomeric excess with a hydride agent formed in situ from LiAlH 4 , (–)-Nmethyl-ephedrine 

and 2-(ethylamino)pyridine (Dyer et al., 1996; Czollner et al., 

1996). Using complex hydrides (NaBH 4 /CeCl 3 , LiAlH 4 /AlCl 3 , etc., –78 °C in 

tetrahydrofuran), narwedine was recently reduced to (–)-galanthamine in high 

yield (99.5%) (Shieh and Carlson, 1995). Carrying out an oxidative cyclisation of 

the biogenetic precursor O-methyl-norbelladine in the presence of extract from 

Narcissus bulbs produced (–)-galanthamine in yields 3 times higher than that 

obtained in the absence of precursor (Bannister and McCague, 1997). In a similar 

route, Chaplin et al. (1997a) used N-methylation of compound 95 before a phenolic 

coupling reaction (30%). Palladium-catalysed debromination produced 

racemic narwedine in 84% yield. Chaplin et al. (1997b) have also patented a 

resolution of racemic bromo-narwedine as dibenzoyltartrate and reduction to 

(–)-galanthamine with L-selectride. 

Galanthamine synthesis by electrochemical oxidation 

of belladine-type amides 

Vlahov et al. (1980a,b) undertook a broad experimental study of the electrochemical 

oxidative transformation of bis-alkoxy- or benzyloxy-substituted bromo-amides 

into derivatives of narwedine and cyclodienones, intermediates in the synthesis of 

1 (Figure 12.13). They synthesised a series of substituted bromoamides 96–98 (R′, 

R″ = Me, PhCH 2 ) and 102. Anodic oxidation of these compounds (+1.30 V in 

MeCN, ≤ 0°C) gave only trace amounts of the desired enones (34 or 49). The 

main compounds isolated were dienones (99–100 or 103) (25–60%), together with

PhCH2O 

MeO 

MeO 

MeO 

PhCH2O 

O 

101 

MeO 

104 

MeO 

PhCH2O 

MeO 

Br 

NMe 

Br 

O 

OMe 

O 

NMe 

OMe 

N 

Br 

105 (R=H) 

106 ( ) 

R=CF 3 

R''O 

R'O 

MeO 

R 

O 

MeO 

RO 

MeO 

NMe 

Br 


unreacted starting materials. Yields were dependent on the character of substitution 

in the substrate molecules. In the case of benzyloxy-substituted compounds, 

yields of the expected compounds reached 25–40%. 

The incomplete transformation of amides 96–98 and 102 to the key compounds 

99–100 and 103 was explained by the existence of starting compounds as a 

O 

96 (R'=R''=Me) 

97 ( R'=R''=PhCH2 ) 

98 ( R'=PhCH2, R''=Me) 

O 

NMe 

Br 

MeO 

R'O 

MeO 

RO 

O 

Br 

99 (R'=Me) 

100 ( R'=PhCH2) 

O 

Br 

NMe 

+ 34 

O 

O 

+ 49 

NMe 

102 103 

R=Me, PhCH2 

R=Me, PhCH2 

PhCH2O 

MeO 

MeO 

OMe 

N 

OMe 

Br 

107 (R=H) 

108 ( R=CF3) 

Figure 12.13 Synthesis of narwedine-type enones and dienones by electrochemical oxidation 

of belladine-type amides (Vlahov et al., 1980a,b, 1984). 

R 

O


mixture of E- and Z-forms. On anodic oxidation of the corresponding N-formyl 

and N-trifluoroacetyl derivatives 105 and 106 (R= CHO and COCF3 ) in the presence 

of MeCN and 2% MeOH, a high rate of conversion of the parent compounds took 

place (80–100%), while the main reaction products were isolated as ketals (107 and 

108, R = CHO and COCF3 ) and the benzylic methylene group underwent oxidative 

methoxylation (Vlahov et al., 1984; Krikorian et al., 1984). The oxidation of amide 

102 in MeCN and 33% MeOH resulted in 30% of methoxylated ketal 104, while 

in the same conditions amide 98 formed methoxycyclohexadienone 101 (Vlahov 

et al., 1984). The electrochemical oxidation reaction was thoroughly studied in 

respect of the effects of substituents and reaction conditions on the reaction products 

and their yields. It was shown that, owing to the high lability of the spirodienones 

produced, they could not be converted to the expected dihydrofuran 

derivatives. 

Synthesis of alkaloids of the galanthamine group using 

hypervalent iodine (III) oxidation agent 

Kita et al. (1998) extended the phenol-coupling reaction using a hypervalent 

iodine (III) reagent for the synthesis of galanthamine-type Amaryllidaceae alkaloids. 

As a result, total syntheses of (±)-sanguinine, (±)-galanthamine, (±)-narwedine, 

(±)-lycoramine and (±)-norgalanthamine were accomplished (Figures 12.14, 12.15 

and 12.16). 

The starting material 3,4-dihydroxybenzoic acid (109), on esterification to 

methyl ester 110 (96%), and following protection of hydroxy groups with 

diphenyldichloromethane as the cyclic diphenylketal 111 and LiAlH 4 reduction 

to the alcohol 112, was brominated with N-bromosuccinimide to afford 113 in 

98% yield (over the three stages). The bromoalcohol 113 was then converted 

(NaOCH 3 – Me 3 SiOSiMe 3 ) to trimethylsilanyl derivative 114 in 56% yield, which 

was oxidised with active MnO 2 to the corresponding aldehyde 115 (86%). Aldehyde 

115 with tyramine (17) in methanol formed the Schiff base 116, which on 

NaBH 4 reduction to norbelladine derivative 117 and acylation (without isolation 

of the latter) with trifluoroacetic anhydride, was converted to amide 118 in 96% 

yield (over the three stages). Phenolic oxidation of 118 with phenyliodine (III) 

bis(trifluoroacetate) in CF 3 CH 2 OH (–40 °C, N 2 ) led to dienone 119 in 36% yield. 

The latter underwent complete de-protection in trifluoroacetic acid medium 

to form enone 120 in 100% yield. Methylation with dimethyl sulphate gave 

N-demethyl-N-trifluoroacetyl-narwedine 121 in quantitative yield (Kita et al., 1998) 

(Figure 12.14). 

The same authors used compounds 121 and 120 (without isolation) for the 

preparation of alkaloids of the galanthamine group (Figure 12.15). Thus, 121 on 

hydrolysis with K 2 CO 3 in aqueous methanol formed N-demethylnarwedine (122), 

which, without isolation, was methylated with formaldehyde and formic acid to 2 

(100%), reduced with L-Selectride (–78 °C in tetrahydrofuran) to 1 (100%), and, 

following hydrogenation over Pd/C, to lycoramine (123) (100%). According to 

another route 121 was hydrolysed to 122 and, without isolation, was reduced with 

L-Selectride to N-norgalanthamine (44) in 82% yield. Sanguinine was synthesised 

as follows: 119 was de-protected and 120, without isolation, afforded 124 in 72% 

yield on treatment with imidazole and tertiary butyldimethylsilyl chloride. Reduction

Ph 

Ph 

O OH 

O 

O OMe 

O OMe 


OH 

OH 

O 

O 

Ph Ph 

O 

O 

Ph Ph 

OH 

OH 

109 110 111 112 

Br 

HO 

O 

OH 

O 

Ph Ph 

N 

Me3Si O 

Ph 

O 

Ph 

O 

Me3Si 

HO 

O 

of 124 with L-Selectride in tetrahydrofuran at –78 °C with following hydrolysis 

gave 125, which on methylation with formaldehyde and formic acid gave sanguinine 

(126) in 68% yield. 

OH 

O 

Ph Ph 

NH 

Me3Si 

Me3Si O 

Ph 

O 

Ph 

HO 

113 114 115 

O 

SiMe3 

N 

O HO 

CF3 

O 

O 

N 

O 

CF3 

HO 

O 

O 

O 

Ph Ph 

N 

O 

CF3 

Me3Si O 

Ph 

O 

Ph 

116 117 118 

MeO 

119 120 121 

Figure 12.14 Synthesis of N,O-didemethyl-N-trifluoroacetylnarwedine (120) and N-demethyl- 

N-trifluoroacetyl-narwedine (121) (Kita et al., 1998). 

O 

O 

N 

O 

CF3


119 

121 

Ph 

Ph 

O 

MeO 

O 

O 

Me O 

SiMe2 

Me 

Me 

O 

O 

122 

44 

O 

O 

N 

O 

NH 

N—COCF3 

CF3 

O 

O 

Ph 

Ph 

127 128 

OH 

Ph 

Ph 

2 

HO 

The dienone 119 was found easily to undergo various transformations (Figure 

12.15). Thus, on trifluoroacetic acydolysis it produced narwedine-type enone 120 

only. On treatment with concentrated HCl or BCl 3 , 119 selectively lost only the 

trimethylsilanyl group with formation of dienone 127. In a medium of 5N HCl in 

O 

1 

HO 

N 

SiMe3 

MeO 

NH 

O 

HO 

O 

HO 

124 125 126 

119 

129 

Figure 12.15 Synthesis of narwedine (2), galanthamine (1), lycoramine (123) and 

sanguinine (126) (above) and reactions of trimethylsilanyl-substituted 

narwedine-type dienone (119) (below) (Kita et al., 1998). 

123 

HO 

O 

O 

HO 

NMe 

N 

NMe 

O 

CF3


ethanol or BBr3 , 119 underwent elimination of the trimethylsilanyl group and 

dienone-phenolic rearrangement, with the formation of apogalanthamine-type 

compound 128. On alkaline hydrolysis with K2CO3 in aqueous methanol, deacylation 

of 119 accompanied cyclisation to crinine-type Amaryllidaceae alkaloids 

129. 

A number of norbelladine-type amides (118) and similar compounds of the 

structure 130–133 were synthesised, and their oxidation process with phenyliodine 

(III) bis(trifluoroacetate) was thoroughly studied (Figures 12.14 and 12.16). 

The main products were found to be trialkylsilanylsubstituted dienones 134 (32%), 

135 (46%), 136 (37%) and 137 (28%). In the case of 118 and 131, the oxidation was 

accompanied by the formation of by-products, dienones 138 (9%) and 139 (12%), 

respectively. The amide 140 on oxidation under similar conditions produced only 

the trimethylsilanyl substituted compound 141, in 26% yield. 

Thus, the most easily removable group for catechol hydroxyls was the diphenylmethylene 

group, and the transformation of dienone to narwedine-type enone 

was successfully achieved only in the case of 119. The enone 120 formed was a 

suitable intermediate for galanthamine type synthesis. 

Synthesis of galanthamine and lycoramine analogues 

By means of photochemical cyclisation of substituted enamidobenzamides spirocyclohexyl-isoquinolines, 

analogues of (±)-galanthamine and (±)-lycoramine have 

been produced (Missoum et al., 1997). Various chemically modified galanthamine 

derivatives (N-C 1 -C 12 -alkylene substituted analogues) have been obtained recently 

as cholinesterase inhibitors (Thal et al., 1997). 

Classical approaches to lycoramine synthesis 

Several multi-step syntheses of (±)-lycoramine have been also published. They are 

not actually biomimetic, and do not include galanthamine or narwedine intermediates. 

Two main strategic approaches were used. The first strategy was based 

on step-by-step construction of a lycoramine system, containing the properly substituted 

benzene nucleus. Thus, Hazama et al. (1968) offered the 23-step synthesis 

from 3-ethoxy-2-hydroxybenzaldehyde in 0.027% total yield. Misaka et al. (1967, 

1968), in a 20-step synthesis from 2,3-dimethoxybenzaldehyde, reached a 0.39% 

total yield. Martin and Garrison (1981, 1982) published the 15-step synthesis from 

2-hydroxy-3-methoxybenzaldehyde, with a 17.35% total yield. A successful 13-step 

synthesis from 3,4-dimethoxycinnamonitrile, in the scheme of Sanchez et al. 

(1984), was realised with over 22.64% total yield. 

In the second strategic approach, the substituted aromatic and cycloaliphatic 

moieties underwent simultaneous modification. Thus, Schultz et al. (1977) have 

used methyl 3-hydroxy-4-methoxybenzoate and 3-ethoxy-2-cyclohexenone as synthones 

in a 16-step synthesis with 8% total yield. Ackland and Pinhey (1987a) used 

cycloaliphatic synthones, 4-ethoxycarbonyl-cyclohexadienones mixture, prepared 

in two stages from 2-trimethylsilyloxy-1,3-butadiene and methylpropiolate, and 

tributyl-(2,3-dimethoxyphenyl)stannane, in a 14-step synthesis with 8% total yield 

(Ackland and Pinhey, 1987b). Parker and Kim (1992) carried out an eight-step 

synthesis starting from cycloaliphatic synthone trans-2-bromo-4-(tert-butyldimeth-


R' 

R' 

HO 

O 

O 

N 

O 

130 (R'=H, R''=Me) 

131 (R'=R''=Me) 

132 (R'=Ph, R''=Et ) 

133 (R'=Ph, R''=Bu) 

HO 

MeO 

MeO 

SiR''3 

CF3 

N 

SiMe3 

O 

CF3 

R' 

R' 

O 

R' 

R' 

ylsilanyl)oxy-cyclohexanone and 2-bromo-3-hydroxy-4-methoxybenzaldehyde (54) 

with a 2.7% total yield. Ishizaki et al. (1993), in a ten-step synthesis from guajacol 

and benzoquinone ethylene monoketal, obtained a 11.3% total yield. The recently 

published synthesis of Essamkaoui et al. (1996) contained only six steps, starting 

from allylcyclohexenylveratrol. 

O 

O 

O 

SiR''3 

134 (R'=H, R''=Me) 

135 (R'=R''=Me) 

136 (R'=Ph, R''=Et ) 

137 (R'=Ph, R''=Bu) 

O 

O 

138 (R '=Ph ) 

139 (R '=Me ) 

140 141 

O 

N 

N 

O 

O 

CF3 CF3 

Figure 12.16 Oxidative cyclisation reactions of trimethylsilanyl-substituted norbelladine-type 

trifluoroacetamides to benzazepine spirodienones (Kita et al., 

1998). 

MeO 

MeO 

N 

CF3 

O 

CF3

CONCLUSIONS 


A comparative analysis of the known schemes of the total synthesis of galanthamine 

has shown that biomimetic methods are preferable over classical synthetic 

approaches because they are carried out in the most economical way, with a minimum 

of steps, with maximum yields and simple realisation. In a single scheme, 

the production of different natural members of the galanthamine series is possible. 

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13 Compounds from the genus Narcissus: 

pharmacology, pharmacokinetics and 

toxicology 

David Brown 

INTRODUCTION 

The genus Narcissus has yielded several useful or potentially useful compounds, 

although just one of them, galanthamine, has been investigated in any great 

detail. As with most modern phytopharmaceuticals, identification, purification 

and investigation of the active principles have only been accomplished after many 

years, perhaps centuries, of folk medicine use as whole plant material or crude 

extracts. For the earliest account of a clinical use of galanthamine one has to go 

back to ancient Greek times. Homer’s Odyssey has it that the attempts of Circe to 

poison Odysseus were foiled when Hermes gave Odysseus an antidotal root drawn 

from the earth, with a black root and milk-like flower, which was difficult for mortals 

to dig up and which the gods called ‘moly’. On the basis of descriptions of their 

symptoms (memory loss, hallucinations and delusions that they had been turned 

into pigs) it has been suggested that the drug which Circe had used to poison 

Odysseus’ men was a powerful, centrally acting, anticholinergic drug called 

stramonium, derived from the common plant Datura stramonium. The story is told 

much more eloquently by Plaitakis and Duvoisin (1983) who postulate that the 

antidote was derived from the common snowdrop, Galanthus nivalis, which contains 

the anticholinesterase, galanthamine. This is based on the description of the 

antidote or ‘moly’ given by Homer and by later Greek texts and recognition that 

both plants (Datura and Galanthus) would have been a resource native to the area. 

The authors state that if all this were true, then it represents the oldest recorded 

use of an anticholinesterase to reverse anticholinergic intoxication. 

Of course, these accounts refer to snowdrop, not narcissus, but galanthamine 

is a derivative common to both. Originally isolated from Galanthus nivalis and used 

clinically under the name ‘Nivalin’, galanthamine can also be derived from 

Narcissus cultivars (Moraes-Cerdeira et al., 1997) and represents the most pharmacologically 

interesting and to date, clinically useful compound, to be derived from 

the genus. There is at the time of writing, considerable interest in galanthamine as 

a potential treatment for Alzheimer’s disease. This chapter reviews the pharmacology, 

pharmacokinetics and toxicology of galanthamine in animals and man. The 

rationale behind its various putative clinical uses is also discussed. Reference is also 

made to another interesting compound, pretazettine, which has been investigated 

as an adjunct to cancer chemotherapy. Clinical (phase 2 and 3) trials with galanthamine 

in Alzheimer’s disease are discussed in the following chapter of this 

volume.

Pharmacology of Narcissus compounds 333 

PHARMACOLOGY OF GALANTHAMINE 

Galanthamine was first isolated in Russia in 1947 and its structure was determined 

in Japan some five years later (Prokurnina and Yakovleva, 1952; Uyeo and 

Kobayashi, 1953). A milestone in the development of galanthamine came in the 

early 1960s, when the compound was first recognised as a reversible inhibitor of 

acetylcholinesterase (Boissier et al., 1960; Irwin and Smith, 1960a). Soon after this, 

evidence was provided that the drug could cross the blood-brain barrier (Nesterenko, 

1964). 

In the 1960s and 1970s, the drug was used in eastern bloc countries for a range 

of neurological disorders (see Clinical Applications, below) but because of the cold 

war, much of the early literature remained obscure. This, and the fact that galanthamine 

was available only as a natural product from limited Bulgarian and Turkish 

sources, explains why Western investigators were slow to appreciate and investigate 

its potential. 

Galanthamine, a tertiary alkaloid (see Figure 13.1), is a selective and reversible 

inhibitor of anticholinesterase, which is an enzyme responsible for the degradation 

of acetylcholine at the neuromuscular junction. The detailed animal pharmacology 

has been reviewed elsewhere (Harvey, 1995). Essentially, galanthamine has 

been shown to inhibit both dog skeletal muscle in vitro and cat and mouse brain 

anticholinesterase in vivo and in vitro in micromolar quantities. When compared 

HO 

H 

O 

O 

CH 3 

N 

CH 3 

N 

O 

OH 

1-adamantyl demethyl galanthamine 

O 

galanthamine 

OCH 3 

O 

O 

HO 

CH 3 

N 

O 

morphine 

OH 

O 

pretazettine 

OCH 3 

H 

OH 

N CH 3 

Figure 13.1 Chemical structures for galanthamine, morphine, 1-adamantyl demethyl 

galanthamine and pretazettine.

334 D. Brown 

with neostigmine the activity of galanthamine against human erythrocyte, human 

serum and dog skeletal muscle cholinesterases was 400–1000 times less potent 

(Boissier and Lesbros, 1962), although functional studies demonstrated a potency 

reduction of only 10–20 times, suggesting that galanthamine may be able to potentiate 

the actions of acetylcholine also. The pharmacological explanation of this 

remains obscure. When the relative potencies of galanthamine, neostigmine and 

pyridostigmine in reversing pancuronium-induced muscle paralysis were studied 

in anaesthetised rats, galanthamine was 30 times less potent than neostigmine and 

12 times weaker than pyridostigmine (Cozanitis et al., 1981). The dose-response 

curve for galanthamine was shallower than those of the other two drugs, indicating 

some as yet unexplained differences in mode of action. Comparative in vitro 

studies have shown that human erythrocyte anticholinesterase inhibition by galanthamine 

is essentially reversible, whereas that of tacrine, a rival treatment for 

Alzheimer’s disease, is not (Tonkopii et al., 1976). In these early experiments, 

tacrine was considerably more potent than galanthamine against animal acetylcholinesterase, 

but in more recent studies, galanthamine was longer acting against 

mouse brain acetylcholinesterase in vivo (Sweeney et al., 1989; Tonkopii and 

Padinker, 1995). 

Galanthamine also inhibits human erythrocyte anticholinesterase selectively, 

having a much smaller effect (from 60 to 100 times less) on butyrylcholinesterase 

both in vitro and in vivo (Thomsen and Kewitz, 1990). This property may be 

important, as more potent inhibition of the latter might increase toxicity in the 

form of neurological side effects – notably prolonged neuromuscular and respiratory 

distress – particularly if the baseline activity of this enzyme is already 

depressed by liver disease or a hereditary defect (Pacheco et al., 1995) or the drug 

is administered long-term, as is likely in Alzheimer’s disease (Thomsen and 

Kewitz, 1990). This observation is also relevant to the selection of drugs for the 

treatment of Alzheimer’s disease, where it is known that physostigmine inhibits 

both acetyl- and butyrylcholinesterase, while tacrine may have more effect on the 

latter than on the former (Thomsen et al., 1991b). Inhibition of erythrocyte anticholinesterase 

may also be used to relate pharmacological activity to therapeutic 

effect, and thus optimise therapy in Alzheimer’s disease. For example, a therapeutic 

window of dosing that stabilises erythrocyte acetylcholinesterase activity at 30–36% 

of baseline has been proposed (Becker et al., 1991). 

Directly relevant to experimental findings in animals is the observation that the 

pro-cholinergic activity of galanthamine was considerable in both human, postmortem 

brain tissue – particularly in the frontal cortex compared with the hippocampus 

– and in fresh cortex samples obtained from operations to remove brain 

tumours (Thomsen et al., 1991a). In these experiments, tacrine was three times as 

potent as galanthamine and physostigmine some 200 times more effective. In spite 

of these differences, the oral daily doses required to achieve equivalent cognitive 

benefit are approximately 30 and 160 mg/day for galanthamine and tacrine, 

respectively. 

Galanthamine has also been shown to inhibit acetylcholinesterase in human 

volunteers and patients with Alzheimer’s disease (Thomsen and Kewitz, 1990; 

Thomsen et al., 1990a, 1991b), although the peripheral (erythrocyte) activity was ten 

times that of the brain activity. Baraka and Harik (1977) found that galanthamine 

(0.5 mg/kg, intravenous (IV)) reversed scopolamine-induced central anticholinergic


syndrome – drowsiness, disorientation, short-term memory impairment – in ten 

healthy volunteers. Although no objective measurements of cognitive function 

were made, electro-encephalogram (EEG) monitoring in two subjects showed 

changes matching the observed changes in consciousness. Scopolamine replaced 

the dominant awake alpha rhythm with slow, disorganised activity; galanthamine 

promptly restored the EEG pattern to normal. 

PHARMACOKINETICS OF GALANTHAMINE 

The reader is referred elsewhere for detailed reviews of the pharmacokinetics of 

galanthamine in species other than man (Harvey, 1995; Bores and Kosley, 1996; 

Bickel et al., 1991a). Animal data are reported here where they serve to amplify 

what is known in man. For a more detailed description of the human data, see 

Kewitz (1997). 

Galanthamine is commonly administered as the hydrobromide salt, and will be 

abbreviated to galanthamine throughout this review, unless other salts were used. 

In general, the pharmacokinetics of galanthamine are first-order and linear over a 

wide dose range. Table 13.1 contains a summary of values determined for key 

pharmacokinetic parameters in man. Galanthamine has been assayed in various 

body fluids using a variety of techniques, including high-performance liquid 

chromatography (HPLC) and enzyme immunoassay. For details of these, see individual 

references. 

Table 13.1 Galanthamine pharmacokinetics: summary of human 

data a 

Parameter Value/comments 

Overall kinetics Linear, two-compartment, 

first order 

Relative oral bioavailability 

(from solution or tablets) 

85–100% 

Time to peak serum level 

52–120 min 

(Tmax ) after tablets 

Absorption half-life (t1/2 abs ) 

Volume of distribution (Vd) 

20 min 

1.76–2.90 litre/kg 

Distribution half-life (t1/2α) Elimination half-life (t1/2β) Mean serum clearance 

6.6–9.6 min 

4.4–8.1 h 

250–340 ml/h/kg 

Mean renal clearance 82–84 ml/h/kg 

Galanthamine excreted 

unchanged in urine 

30–50% 

Dose excreted in first 24 hours 

Metabolites (see Figure 13.2): 

60% 

O-demethyl galanthamine (iii) 20% as glucuronide 

N-demethyl galanthamine (iv) 5% 

Epigalanthamine (v)

336 D. Brown 

Absorption 

Galanthamine had almost complete (85–100%) oral bioavailability in man, when 

formulated either as a solution or tablets (Mihailova et al., 1989; Bickel et al., 

1991b). In one study (Bickel et al., 1991b), the time to peak plasma concentration 

(T max ) was shorter with a solution (15 minutes against 52 minutes). The half-life 

of the absorption process was 20.4 minutes. In another study (Mihailova et al., 

1989), absorption from oral tablets was slower, with a T max in healthy volunteers 

of approximately 120 minutes. The bioavailability data indicate that there is no 

significant first-pass effect. 

Therapeutically relevant drug concentrations, corresponding to 30–60% inhibition 

of erythrocyte acetylcholinesterase (as recommended by Becker et al., 1991) 

were reached within 30–44 minutes of administration. 

Distribution 

Data from studies in anaesthetised cats indicate that galanthamine has a large 

volume of distribution but does not bind to plasma proteins significantly 

(Mihailova et al., 1985). Single dose (0.3 mg/kg, IV) studies of galanthamine have 

been carried out in eight female patients undergoing gynaecological surgery 

(Westra et al., 1986). Galanthamine was given at the end of surgery to reverse 

pancuronium-induced neuromuscular blockade. Serum levels were determined 

by HPLC. Mean peak levels of 543± 47 ng/ml were observed at approximately 

2 minutes. The serum level versus time curve showed biexponential decay after 

the maximum was reached, indicating extensive distribution in a two-compartment 

model. A mean distribution half-life (t 1/2 α) of 0.11 hours and an elimination halflife 

(t 1/2 β) of 4.4 hours were calculated. The latter was considerably longer than 

the half-lives of neostigmine (80 minutes) and pyridostigmine (46 minutes). The 

volume of distribution at steady state was large (mean: 1.76 litre/kg, with a 95% 

confidence interval (CI) of 1.22–2.30 litre/kg), indicating accumulation by various 

tissues. The mean total serum clearance was 322.2 ml/h/kg and mean renal clearance 

was 81.6 ml/h/kg. 

Mihailova et al. (1989) administered single doses of 10 mg (0.11–0.18 mg/kg) 

subcutaneously or orally, in an open crossover fashion, to eight male volunteers. 

By these routes, the T max was prolonged (2 hours in each case) and peak serum 

levels were 1.1–1.5 µg/ml and 1.0–1.4 µg/ml, respectively. The authors concluded 

that the subcutaneous and oral formulations were essentially bioequivalent. These 

values are higher than one might expect on the basis of the results of Westra et al. 

(1986) who gave, on average, three times the dose of galanthamine intravenously. 

The discrepancy is difficult to explain on the basis of different assay methods 

employed in the two studies alone. Perhaps there are as yet unexplained differences 

in age, sex or clinical status that determine the pharmacokinetics of galanthamine. 

The t 1/2β values were 5.7 and 5.3 hours after oral and subcutaneous 

doses, respectively. 

Thomsen et al. (1990b) reported a pharmacokinetic study in one Alzheimer’s 

patient and one volunteer. Repeated daily doses of 30–50 mg galanthamine 

resulted in steady state plasma concentrations of 50–150 ng/ml. In the volunteer, 

IV injections of 10 mg of the drug produced a peak plasma concentration of


182 ng/ml. The half-life was reported to be 8–10 hours. These values are closer to 

those reported by Westra et al. (1986). In a further study in eight male volunteers, 

Bickel et al. (1991b) observed a maximum plasma concentration of 30–60 ng/ml 

after oral administration of 10 mg galanthamine and 50–100 ng/ml after the same 

dose given by constant rate IV infusion over 30 minutes. Galanthamine showed 

linear, first order pharmacokinetics. The mean values for t1/2α and t1/2β were 0.16 

hours and 5.68 hours, respectively. The mean total clearance was 340 ml/h/kg and 

renal clearance was 84 ml/h/kg. A steady state volume of distribution of 2.64 litre/kg 

(95% CI: 2.41–2.90 litre/kg) was calculated. Twenty-five percent of the dose was 

excreted unchanged in the urine. Little evidence of metabolism was found: negligible 

amounts of epigalanthamine and galanthaminone were detected in blood 

and urine. 

Most recently, Kewitz (1997) reported data from a 15-week clinical trial of 

galanthamine in 34 patients with Alzheimer’s disease, whose mean age was 75 

years. A mean volume of distribution of 2.9 litre/kg, a half-life of 8.1 hours and a 

plasma clearance of 250 ml/h/kg were calculated. As might be expected and 

certainly should be remembered, clearance was 30% less in this elderly population 

than in healthy young volunteers. 

There is no direct in vivo evidence that galanthamine itself enters the human 

brain. However, examination of its structure (see Figure 13.1) reveals a tertiary 

nitrogen atom in common with many molecules that can penetrate the bloodbrain 

barrier. Indirectly, there is biochemical evidence from experiments in rats 

(Mihailova and Yomboliev, 1986). One and 3 mg/kg intravenous doses produced 

peak levels of 5 and 6.3 µg/g of brain tissue, respectively, 10–15 minutes after 

injection, declining to about 10% between 1 and 3 hours. In mice, parenteral 

administration of galanthamine resulted in accumulation in brain, liver and 

kidney tissue (Bickel et al., 1991a). 

Correlation of galanthamine pharmacokinetics with erythrocyte 

cholinesterase inhibition 

The inhibition of acetylcholinesterase in human erythrocytes correlates closely 

with the pharmacokinetics of galanthamine. A median maximal value of 53% 

inhibition was reported after a single, intravenous dose of 10 mg (Bickel et al., 

1991b). The in vitro and ex vivo concentration responses were essentially identical, 

indicating that galanthamine alone, and not its metabolites, was responsible for 

inhibition of cholinesterase. In a previous study (Thomsen et al., 1990a), erythrocyte 

acetylcholinesterase activity returned to normal some 30 hours after both 

single dose and chronic administration at therapeutic doses (10–40 mg daily), illustrating 

the reversible nature of the inhibition of the enzyme by galanthamine. 

Kewitz (1997) recently presented data to show the relationship between the 

serum level of galanthamine, enzyme inhibition in erythrocytes and central 

nervous system (CNS) effects. Simultaneous EEG recordings were made in a 

healthy young male volunteer, during an IV infusion of 15 mg galanthamine. The 

power density of the alpha-one frequency band of the EEG at the occipital region 

began 7 minutes after starting the infusion, when 40% of the enzyme activity had 

been blocked. Power density fluctuated in synchrony with enzyme activity as the 

galanthamine levels fell after the infusion was stopped.

338 D. Brown 

Metabolism and excretion 

Clearly, the metabolism of any drug that is used in an elderly population needs to 

be characterised, given the potential effects of age on liver function. 

In the study by Westra et al. (1986), approximately 30% of the intravenous 

galanthamine dose was excreted in the urine unchanged. Biliary excretion was 

negligible and there was no evidence of conjugation with glucuronide or sulphation. 

Mihailova et al. (1989) detected the metabolites epigalanthamine (the stereo 

isomer of galanthamine – see Figure 13.2) and galanthaminone in plasma and 

urine; these metabolites possess only very weak anticholinesterase activity (


on the excretion of galanthamine or its metabolites in bile or the intestinal tract. 

The notion that galanthamine is not extensively metabolised is also supported by 

animal data (Harvey, 1995). 

More recently, Bachus (1995) showed that in male volunteers, 20% of a dose of 

galanthamine appeared in urine as O-demethylgalanthamine glucuronide, 5% as 

N-demethylgalanthamine and < 2% as epigalanthamine resulting from the intermediate 

metabolite galanthaminone, which itself was present in traces only. 

O-demethylgalanthamine is 3–10 times more active than galanthamine, but rapid 

glucuronidation reduces this by a factor of at least one hundred. All the other 

galanthamine metabolites identified thus far have little anticholinesterase activity. 

O-demethylation is catalysed by the cytochrome isoenzyme CYP2D6. This knowledge 

should allow the avoidance of concomitant drugs that are also metabolised 

predominantly by this isoenzyme (such as beta-blockers, haloperidol, morphine, 

phenothiazines and some tricyclic and selective serotonin reuptake inhibitor 

antidepressants) and thus decrease the potential for drug interactions through 

metabolism induction or inhibition. Fluoxetine, haloperidol, morphine and quinidine 

are known to inhibit CYP2D6, but the problem may be largely theoretical. 

Co-adminstration of therapeutic doses of quinidine, a potent inhibitor of CYP2D6, 

to four healthy volunteers completely suppressed O-demethylation but led to only 

a modest (20%) rise in galanthamine serum levels. Kewitz (1997) claimed that this 

rise was unlikely to cause toxicity and concluded that co-administration of other 

drugs inhibiting the same enzyme would do little damage. Neither would problems 

be likely in poor metabolisers, deficient in CYP2D6 (5–10% of Caucasians and 1% 

of Asians), however caution is advisable. No comment was made on the likely outcome 

if other interacting drugs were used in poor metabolisers or in those with 

kidney dysfunction. Potential inhibitors of the isoenzyme CYP3A4 should also be 

viewed with caution, as this enzyme also plays a part in galanthamine metabolism. 

Comparison with other relevant drugs 

Galanthamine pharmacokinetics appear to be more attractive than those of some, 

but not all, other anticholinesterases currently under investigation for the treatment 

of Alzheimer’s disease. Physostigmine has poor oral bioavailability (approx. 10%) 

and a short half life (

340 D. Brown 

were internationally inaccessible. Western research conducted at this time merely 

paralleled and often duplicated the eastern European effort, rather than building 

upon it. Add to this the fact that early preparations of galanthamine were obtained 

by liquid carbon dioxide extraction of plant tissue and were therefore only available 

in small and often unreliable quantities of uncertain purity. Although a full synthesis 

of galanthamine was published in 1960 (Barton and Kirby, 1960), full-scale industrial 

synthesis was shown to be possible at a much later date, implying that a supply 

of predictable and reproducible purity was not used in early trials. 

Historically, the considerable interest in galanthamine is illustrated by the fact 

that patents have been filed for its use in conditions as diverse as nicotine addiction, 

alleviation of benzodiazepine side effects, male erectile dysfunction, chronic 

fatigue syndrome, bipolar and panic disorders and acute mania (Mucke, 1997). 

There is a rational basis to many of the therapeutic uses to which galanthamine 

has been put. They may be divided into those, where, reversal of excessive anticholinergic 

activity is required, and those where, cholinergic activity is deficient. 

Uses of galanthamine where reversal of excessive 

anticholinergic activity is required 

Reversal of drug-induced paralysis in anaesthetics 

The ability of galanthamine to reverse tubocurarine-induced muscle paralysis was 

discovered almost half a century ago (Mashkovskii, 1955). The drug has been used 

extensively to reverse the effects of curare and other muscle relaxants after surgery 

in eastern European countries, but published reports are largely anecdotal (see 

review by Paskov, 1986). Use has been rare in the West, where reports are fewer 

but better documented (Mayrhofer, 1966; Wislicki, 1967; Cozanitis, 1971; Baraka 

and Cozanitis, 1973). Typically, an intravenous dose of galanthamine is given to 

induce spontaneous respiration and awakening. Sometimes a second dose may be 

given, 5–10 minutes after the first. It is claimed that galanthamine has fewer cholinergic 

side effects than neostigmine, thus reducing the need for compensatory 

atropine, and that galanthamine speeds recovery from respiratory depression 

caused by morphine analgesia, but these effects are not fully substantiated. 

For example, Cozanitis (1971) reported the successful use of galanthamine 

given intravenously in graded doses to a maximum of 20 mg in 40 surgical 

patients, to reverse the effects of alcuronium, pancuronium, gallamine and 

tubocurarine. The recovery rate was slower than that usually seen with neostigmine; 

atropine was not necessary. Side effects included increased pulse rate in 

most patients, increased salivation, dizziness and injection site reactions in some 

patients. Galanthamine was judged to be approximately 20 times less potent than 

neostigmine (Baraka and Cozanitis, 1973). 

Uses of galanthamine where cholinergic activity is deficient 

Research effort in this area has focused almost exclusively on investigating galanthamine 

as a potential treatment for Alzheimer’s disease (AD). Phase 2 and Phase 3 

clinical trials with galanthamine in this indication are reviewed in the following 

chapter of this volume.


Early work in animals 

Galanthamine reversed the amnesia induced in mice by the injection of the anticholinergic 

agent, scopolamine (Chaplygina and Ilyutchenok, 1976). The drug 

was also shown to reverse the cholinergic deficit induced in rats by pre-treatment 

with the neurotoxin ibotinic acid (Sweeney et al., 1988). In a notable series of 

experiments, Sweeney and co-workers produced lesions in the nucleus basalis 

magnocellularis of mice; this region is a major site of cholinergic enervation to the 

fronto-parietal cortex, and lesions here produce significant deficits in choline 

acetyltransferase activity which are linked to deficits in spatial memory. Intraperitoneal 

administration of galanthamine attenuated these deficits (Sweeney et al., 

1989), and improved memory in swim tasks (Sweeney et al., 1988) and passive 

avoidance tests (Sweeney et al., 1990); repeated doses remained effective, indicating 

that tolerance to the drug did not develop. Control animals given sham lesions 

showed impaired performance. At relatively high doses (1–4mg/kg) galanthamine 

impaired the performance of control animals markedly, although an explanation 

for this was not forthcoming (Sweeney et al., 1990). 

Other workers confirmed some of this work: for example, Yonkov and 

Georgiev (1990) showed that galanthamine enhanced the retention of learned 

behaviour in both active and passive avoidance tests in rats. Chopin and Brierly 

(1992) showed that galanthamine was capable of reversing scopolamine-induced 

poor performance in a passive avoidance test in rats at very similar doses to 

tacrine (0.3–10 mg/kg administered intraperitoneally), a measure of the effect on 

memory. The doses used in these experiments were far in excess of those used in 

man to treat AD. Fishkin et al. (1993) showed that intraperitoneal galanthamine 

(1.25–5 mg/kg) significantly attenuated scopolamine-induced learning and memory 

deficits in rats, compared to controls as measured by T maze and Morris 

water maze experiments. 

An interesting hypothesis, relevant to the pathophysiology of AD, is that in addition 

to inhibition of acetylcholinesterase, inhibitors of this enzyme may activate 

normal processing of the amyloid precursor protein which accumulates in the 

disease producing amyloid plaques which are a striking feature of post-mortem 

brain samples (Giacobini et al., 1996). Whether or not galanthamine provides such 

neuroprotection is unknown, and definitive data will come only from long-term 

trials in man, where biopsy or post-mortem samples are available for study. 

Early work in man 

Early reports from eastern Europe claimed that recovery of consciousness after 

surgical anaesthesia was faster if galanthamine was used instead of neostigmine as 

a curare antagonist (Paskov, 1986). The mechanism remains obscure; it might be 

due to a direct central stimulant action or the ability to antagonise the action of 

opiate analgesics. The results from a study in conscious volunteers indicate that 

galanthamine may act as a mild central stimulant (Cozanitis and Toivakka, 1991). 

Galanthamine crosses the blood-brain barrier readily, and because of this and 

the in vitro data discussed above, it was soon proposed as a potential treatment for 

brain diseases where cholinergic transmission appeared to be deficient – notably 

AD. In this disease, destruction of cholinergic neurones in the brain, notably the

342 D. Brown 

cortex and hippocampus, results in reduced levels of acetylcholine which are 

thought to be associated with loss of memory and disrupted cognition. Drugs that 

preserve acetylcholine by inhibiting its destruction by cholinesterase, such as 

neostigmine, rivastigmine, tacrine, metrifonate and galanthamine, can reduce 

AD symptoms and delay, but not halt, disease progression (Mucke, 1997). Several 

small trials and what amount to anecdotal reports provide an impression that 

galanthamine is useful in this respect and, because of this early promise, several 

larger and more carefully controlled trials are underway. This work is reviewed in 

the next chapter of this volume. Specific, non-AD applications are described 

below. 

Neurological injury 

Paskov (1986) reviews the use of galanthamine to treat a wide range of neurological 

disorders. Most of the evidence is anecdotal and unclear, but it is logical to 

expect that if the neurological injury involves a cholinergic pathway resulting in a 

reversible deficit or imbalance of acetylcholine, then galanthamine may be of use. 

Reversal of pathological paralysis 

In the 1960s, galanthamine was used, again in eastern European countries, to 

assist recovery from paralysis associated with poliomyelitis, neuromuscular disorders 

or neuromuscular diseases. Trigeminal neuralgia was managed with oral and 

intramuscular administration and occasionally by infiltration. Doses of 15–25 mg/ 

day were typically used (Kilimov, 1961a). Galanthamine was reported to produce 

motor function improvement in 15 of 25 children with muscular dystrophy (Pernov 

et al., 1961) and in patients with myasthenia and neuromuscular dystrophy (Pestel, 

1961). Sixteen of 20 patients with brain injuries (not specified) showed improvement 

on galanthamine (Kilimov, 1961b). Revelli and Grasso (1962) noted varying 

degrees of benefit in 49 of 52 patients suffering from post-febrile polio during a 

3-month trial of the drug at doses of 2.5–10 mg/day. Gopel and Bertram (1971) 

reported the use of galanthamine in an heterogeneous group of patients studied 

over a 30-month period, using subcutaneous doses of galanthamine in the range 

1.25–25 mg daily. Beneficial effects were seen in the following disorders, some of 

which were resistant to conventional therapy: facial paralysis (22 of 22); peripheral 

neuropathy (38 of 46); paralysis of central origin (11 of 12); progressive myodystrophy 

(1 of 2); poly/dermatomyositis (3 of 3) and multiple sclerosis (4 of 4). The 

authors stated that the drug was well tolerated. 

Gujral (1965) studied 100 patients with post-polio spinal paralysis, seven with 

muscular dystrophy and two with facial paralysis due to lower motor neurones. 

Galanthamine was administered as a series of courses of 40 subcutaneous injections, 

repeated after a 6- to 8-week interval, up to a maximum of three courses per 

patient. Total daily doses were related to age. Drug treatment was combined with 

physiotherapy; this may have confounded the results, as physiotherapy was 

already proven to be effective and was not administered uniformly. In any event, 

the results were far from conclusive. Only modest number of patients showed 

improvement with galanthamine. The author concluded that the drug was most 

likely to have a beneficial effect in early cases of post-polio paralysis (improvement


in muscle tone and isometric contractions) and at an early, rather than late, phase 

of treatment (5–15 days of the first course). No improvement was seen where the 

muscle showed zero power from the onset or in chronic cases. Very little improvement 

was seen in four of the seven cases of pseudo-hypertrophic muscular 

dystrophy and none in the remaining patients. Some improvement in the two 

children with facial paralysis was noted. Galanthamine was described as being well 

tolerated; mild reactions included salivation, nausea and abdominal pain that 

resolved on decreasing the dose. 

Improvement in motor function was observed when galanthamine was used as 

part of a programme of therapies used to treat polyneuropathy-associated Guillain- 

Barre syndrome (Kuyumdzhieva et al., 1996). 

Use in poisoning 

Reports in animal, and later in human, studies, of the reversal of respiratory 

depression caused by opiate analgesics are tempered by observations that cholinergic 

side effects are likely at the doses required and that only temporary reversal 

is achieved (Tassonyi et al., 1976). However, galanthamine has been used to 

reverse CNS effects in several cases of drug overdose, including scopolamine 

(Cozanitis, 1977), Mandrax (a combination of diphenhydramine, a drug with 

marked anticholinergic properties and methaqualone, an hypnotic), and dextromoramide 

(a narcotic analgesic related to methadone, which caused marked 

respiratory depression) (Cozanitis and Toivakka, 1974). 

Analgesia 

Poultices of the leaves and bulbs of the Amaryllidaceae have been reported in 

herbal lore for hundreds of years as being used to treat painful neurological 

conditions such as facial neuralgia; this treatment was said to provide analgesic 

and curative effects (Rainer, 1997). 

Despite its apparent ability to reverse the respiratory depression seen with 

morphine-like analgesics, galanthamine does not appear to antagonise opiateinduced 

analgesia. Indeed, the drug is structurally related to morphine (see 

Figure 13.1). Evidence from standard laboratory tests suggests that the drug may 

have mild analgesic properties (Cozanitis et al., 1983). Suggestions that galanthamine 

may stimulate opioid receptors directly needs substantiation by direct, 

radioligand binding studies. 

Migraine 

Ikonomoff (1968) was the first to report the use of galanthamine in the management 

of migraine patients. Of 140 cases involving galanthamine (in the form of the 

commercially available product ‘Nivalin’), 25 (18%) were headache free for 6 

years, and a further 64 (46%) had shown some improvement. In 42 patients, spontaneous 

epistaxis resolved and in 56 patients, subcutaneous ecchymosis ceased 

when galanthamine was used. Twenty-two of 59 female patients with dysmenorrhoea 

accompanying their migraine experienced resolution of period pains. 

Neostigmine was found to be superior in a similar range of patients. The author

344 D. Brown 

reasoned that the efficacy of these two drugs could not be explained on the basis of 

their anticholinesterase activity alone, and suggested (but presented no evidence 

for) a range of actions, including beneficial effects on blood vessel tissue and a 

reduction of capillary permeability. Treatment, given twice daily, with an 

optimum total daily dose of 250 mg, was well tolerated, and no patient had to 

discontinue therapy because of side effects. 

Mania and schizophrenia 

Disturbances in the balance between neurotransmitters may be manipulated to 

improve a lot of patients suffering from mania and schizophrenia where, complex 

imbalances often occur. Snorrason and Stefansson (1991) have used this argument 

to justify treating manic patients with galanthamine. There is minimal detail in 

this report but galanthamine, 10 mg, three times a day, was observed to improve 

symptoms rapidly when given to a 74 year old lady who was unresponsive to lithium 

and where other agents were contraindicated because of neuroleptic malignant 

syndrome. The patient’s condition deteriorated when galanthamine was 

stopped. The authors referred to ten other patients where the drug was of benefit. 

Galanthamine has been associated with a slow improvement in psychomotor function 

deficits in 18 of 30 patients with schizophrenia over a 3- to 4-week period 

(Vovin et al., 1991). 

Raised intraocular pressure 

One paper mentions experiments where, galanthamine reduced intraocular pressure 

when applied in an eye drop formulation to rabbits (Agarawal and Gupta, 

1990). The effect was slow and peaked at 2 hours. Physostigmine is an established 

drug for the treatment of glaucoma and it is interesting that galanthamine might 

also prove useful. There are no published trials of this use in man. 

Chronic fatigue syndrome (CFS) 

Several symptoms of CFS such as sleep disturbances, poor concentration and generalised 

fatigue are similar to the side effects of anticholinergic drugs; these can be 

reversed with physostigmine, a cholinesterase inhibitor similar to galanthamine. 

The latter has been shown to shorten rapid eye movement (REM) latency, increase 

REM density and reduce slow wave sleep, modify poor concentration, memory 

disturbances and reverse behavioural changes seen in Alzheimer’s disease. In 

addition, galanthamine can elevate plasma cortisol levels, whereas patients suffering 

from glucocorticoid deficiency have similar symptoms to those with CFS. 

These observations have prompted suggestions that galanthamine may be of use 

in this syndrome. Thirty-three patients were enrolled in an 8-week, double-blind, 

placebo-controlled study involving galanthamine (Snorrason, 1993). Patients in 

the active group received total divided, daily doses of 10–50 mg. The following 

symptoms were improved significantly in the galanthamine group: sleep disturbances, 

fatigue, myalgia, anxiety, immediate and delayed recall; improvements in 

cognition were also observed. Similar results were published three years later by 

the same author (Snorrason et al., 1996), highlighting marked improvements in


sleep disturbances as a potential benefit of the drug. Nausea was the most common 

side effect, reversible on drug withdrawal. 

TOXICOLOGY AND TOXICITY OF GALANTHAMINE 

Toxicological data derived from animals appear to be of limited use in predicting 

safety in the intended human population, but some data are of interest. The 

reader is referred to reviews on this topic for further information (Harvey, 1995; 

Mucke, 1997). 

Animal toxicology 

Acute toxicity is manifested mainly as exaggerated cholinergic effects that may be 

reversed with atropine. Death occurs through respiratory depression, whereas 

cardiac function is relatively unaffected. An acute LD 50 of 5.2 mg/kg was determined 

in mice after IV administration (Friess et al., 1961). The latter may have been an 

underestimate, as a value of almost double this was obtained when galanthamine 

was co-administered with 4-aminopyridine, a drug that enhances the release of 

acetylcholine from nerve terminals (Micov and Georgiev, 1986). A chronic toxicological 

study of the combination, with galanthamine doses up to 2 mg/kg daily, 

produced no remarkable changes in blood or major organs (Micov and Georgiev, 

1986). As expected, the oral LD 50 was higher than the IV value, at 18.7 mg/kg 

(Umarova et al., 1965). Chronic oral doses (0.5 mg/kg/day) were shown to reduce 

respiratory volume and to desynchronise EEG patterns in rabbits, but not in other 

species, at this low dose. Maternal and embryo-toxicity were observed in rats and 

rabbits at doses of 10% of the LD 50 (Paskov, 1986). 

Toxicity in man 

As an inhibitor of acetylcholinesterase, one might expect cholinergic side effects 

to be a feature of the side effect profile of galanthamine. Indeed, these have been 

reported following human consumption when narcissus bulbs have been mistaken 

for onions, or the leaves and flowers have been eaten (Vigneau et al., 1984). Early 

reports of successful use in anaesthesia to reverse neuromuscular junction blocking 

agents mentioned few autonomic side effects – which is probably a reflection 

on their anecdotal and enthusiastic nature. More recent and carefully controlled 

studies confirm the suspicion that autonomic effects are indeed to be expected, 

but that these are dose related. Altered electrocardiogram (ECG), blurred vision, 

hypersalivation, nausea, vomiting and dizziness have been noted in conscious 

volunteers given 20 mg doses. Interestingly, galanthamine produced mild eosinophilia 

after injection. According to the authors, this might explain the local tissue 

reaction noted on injection of the drug (Cozanitis and Toivakka, 1971). No 

changes in blood sugar were observed in conscious volunteers (Cozanitis and 

Toivakka, 1971; Riemann et al., 1994) and in patients after surgery (Cozanitis 

et al., 1973b). At a relatively high dose of 25 mg, three out of nine surgical patients 

had sufficient bradycardia to require reversal with atropine (De Angelis and Walts, 

1972).

346 D. Brown 

Galanthamine causes bradycardia and might be expected to have a negative 

effect on blood pressure. However, when the drug has been tested in animals at 

larger doses than those used therapeutically in man, species-specific differences 

have been observed. A significant fall in blood pressure was noted in anaesthetised 

dogs (Irwin and Smith, 1960a,b), but a significant rise was noted in anaesthetised 

rats (Chrusciel and Varagic, 1966). In the latter experiment, physostigmine but 

not neostigmine produced a similar effect, suggesting a central mechanism, but 

the effect was not blocked by autonomic ganglion blocking drugs such as hexamethonium. 

Galanthamine-induced hypertension was attenuated by drugs capable 

of blocking the central action of the drug, such as adrenoreceptor antagonists, 

nicotine and atropine. Galanthamine produced no hypertensive response in 

pithed rats. With the exception of the lack of effect of the sympathetic ganglion 

blockers, the evidence points toward a central stimulant action, at least in anaesthetised 

rats. 

Mild bradycardia was noted as a side effect in a trial of galanthamine in 14 

surgical patients. A dose of 20mg produced a reduction in mean pulse rate to 69 

from 72 beats per minute (Cozanitis et al., 1973a), but this had no appreciable 

effect on blood pressure. Six patients had more marked bradycardia, down to 55 

beats per minute, associated with minor arrhythmias. When the drug was administered 

with atropine, mild bradycardia persisted, but only two patients showed 

anything like the more dramatic falls in pulse rate described above. In a trial by 

Khmelevsky and Gadalov (1980), galanthamine without atropine produced significant 

bradycardia in just two of 40 surgical patients. As with other cholinergic 

drugs, one should be alert to the possibility of cardiac complications in patients 

with pre-existing disease and in those suffering from Parkinson’s disease (often 

co-morbid with AD) where other treatments may interact. Very limited evidence 

suggests that galanthamine may be administered with selegiline or muscarinic 

receptor blockers with appropriate caution (Losev and Kamenetski, 1985; 

Rainer, 1997) but there is no evidence to suggest that co-administration is safe 

with other drug classes that are likely to interact. The effects of galanthamine are 

likely to be additive with other anticholinesterases and cholinergics (including 

those with cholinergic side effects such as the non-selective mono-amine oxidase 

inhibitors) and the drug would be expected to antagonise and be opposed by anticholinergic 

drugs such as the antiparkinsonian agents. Thus, it is feasible that a 

centrally-acting anticholinergic drug might oppose the efficacy of galanthamine in 

AD, while one which acts peripherally might reduce unwanted side effects. 

As with other drugs used in anaesthesia, it is important to know if galanthamine 

affects respiration. Cozanitis et al. (1972) used cine-bronchography to research the 

effect of galanthamine on the bronchial tree in five asthmatic volunteers. Parenteral 

doses of 20 mg produced no suspicious changes and blood gas composition 

was not altered. 

The ability of galanthamine, in common with some other cholinergic drugs, to 

stimulate raised serum levels of cortisol was noted quite early on in the use of the 

drug in anaesthesiology (Cozanitis et al., 1973b; Cozanitis, 1974). Typically, a 

single 20 mg dose of galanthamine produced a 48% increase in plasma cortisol 

from 0.54 to 0.8 µM/litre. No effect was observed with neostigmine, leading the 

authors to suggest that galanthamine was working centrally. A later study by the 

same group (Cozanitis et al., 1980) demonstrated that galanthamine was capable of


stimulating the release of adrenocorticotrophic hormone (ACTH) in eight surgical 

patients. Again, neostigmine failed to produce this effect. Although a central 

action seems likely, the exact mechanism remains obscure. 

No changes in liver function tests were observed in these early studies. Reviewing 

clinical trials to date, there is virtually no evidence that galanthamine has any liver 

toxicity. 

As a result of its proposed use in patients with AD, attempts have been made to 

characterise the CNS toxicity of galanthamine. Because of a previous observation 

that galanthamine was a mild CNS stimulant (Cozanitis and Toivakka, 1971), 

the same authors investigated the epileptogenic potential of galanthamine in 18 

epileptics controlled with phenytoin (Cozanitis et al., 1973b). Six patients showed 

increased abnormal EEG activity with one patient showing marked increases in 

spike and paroxysmal wave activity; no seizures were precipitated. EEG changes 

indicating CNS stimulation have also been reported in cats (Kostowski and 

Gumulka, 1968), but a series of experiments in rabbits indicated that high doses 

(1 mg/kg) could suppress epileptic activity induced by instillation of penicillin into 

the dorsal hippocampus (Losev and Tkachenko, 1986). 

Onset of REM sleep is known to be controlled by cholinergic mechanisms in the 

brain stem. Studies have shown that galanthamine reduces the latency of REM 

sleep and the overall duration of slow-wave non-REM sleep after single, 10 or 15 mg 

doses of galanthamine in healthy volunteers (Rieman et al., 1994). The result is 

typical of other cholinergic agents and mimics some of the sleep abnormalities 

seen in major depressive disorders. The relevance of this to the use of galanthamine 

in AD, where patients can become clinically depressed, is unclear. 

Safety data from clinical trials in Alzheimer’s disease 

The therapeutic activity of galanthamine, studied in a variety of clinical trials 

involving patients with AD, is discussed in the next chapter. Side effects were not 

reported uniformly and in some cases it is not clear whether they were omitted 

from the report or did not occur. The following side effects have been observed 

under clinical trial conditions. 

In a small trial involving four Alzheimer’s patients, who received galanthamine 

15, 30 and 45mg daily in an escalating regimen over 2–3 months, galanthamine 

was well-tolerated at the two lower doses, but only one patient could tolerate 

45 mg/day; the others experienced agitation and insomnia (Thomsen et al., 1990a). 

Dal-Bianco et al. (1991) described a preliminary study where six Alzheimer’s 

patients were given galanthamine (30–50 mg daily) for up to 16 months without 

apparent adverse effects. However, this too was a small trial; larger studies show 

that at therapeutic doses, cholinergic effects are an annoying problem with galanthamine. 

They are usually mild, consisting of nausea and vomiting, which can be 

minimised by careful, stepwise upward titration over 1–2 weeks. Conventional 

antiemetics such as metoclopramide or domperidone may be useful in short 

courses lasting a few days. 

In a placebo-controlled trial, 44 patients received galanthamine 20–50 mg/day 

in two or three divided doses for 10 weeks (Kewitz and Davis, 1994). The most 

common adverse events were nausea (16%) and vomiting (19%). Abdominal pain, 

diarrhoea, agitation and dizziness each occurred in 4% of patients. Kewitz et al.

348 D. Brown 

(1994) go on to mention some patients with light-headedness, agitation and sleep 

disturbances (incidences were not reported), but there were no withdrawals due 

to adverse effects. Liver function tests and serum creatinine remained normal 

throughout. 

DERIVATIVES OF GALANTHAMINE 

As discussed previously, galanthamine is not an ideal drug; cholinergic side effects 

such as nausea and vomiting occur in significant number of patients at doses 

accepted as therapeutic. Consequently, there has been a degree of research interest 

in developing compounds with fewer side effects, and reports of semi- or 

wholly synthetic galanthamine derivatives are beginning to appear. The complete 

chemical synthesis of galanthamine was achieved and reported almost 40 years 

ago (Barton and Kirby, 1960) and an economic production process has recently 

been developed which should reduce the cost and guarantee a supply of parent 

compound with a consistent level of purity (Holton et al., 1998). Bores and Kosley 

(1996) provide an extensive review of attempts to improve upon the safety, 

efficacy and pharmacokinetic profile of galanthamine through the synthesis of 

analogues. Preclinical assessment of the 6-ester derivatives of 6-demethylgalanthamine 

showed most promise, particularly the adamantyl ester (see Figure 13.1). 

This compound, like other 6-ester derivatives appears to act as a prodrug. After 

oral administration and absorption, rapid hydrolysis takes place, presumably by 

non-specific blood and tissue esterases, yielding the potent acetylcholine esterase 

inhibitor, 6-demethylgalanthamine – see Figure 13.2 (Bores et al., 1996). This 

appears to be five times as potent as galanthamine at inhibiting mouse brain 

acetylcholinesterase (Han et al., 1991, 1992), and compared with galanthamine it is 

a more selective inhibitor of acetylcholinesterase rather than butyrylcholinesterase. 

Galanthamine had better overall bioavailability than any of the ester 

analogues, including the adamantyl ester, in rats; but the adamantyl ester was 

judged to have the most favourable pharmacokinetics (i.e., slower onset and 

sustained levels of 6-demethylgalanthamine, combined with the greater potency of 

the latter towards acetylcholinesterase). The compound also had a higher oral 

therapeutic index, higher brain levels and greater activity in vivo, as demonstrated 

by greater efficacy at causing hypothermia (Bores et al., 1996). The authors 

concluded that because of its composite profile, including duration of action, oral 

therapeutic index and pharmacokinetics, the adamantyl ester was the best therapeutic 

candidate for the treatment of AD. This hypothesis awaits confirmation 

from experiments with the compound in higher species and man. 

PRETAZETTINE 

Compounds in Narcissus species have joined the long line of phytochemicals that 

have been investigated for their anti-tumour activity. Several early reports, most of 

them originating from a single research group in Honolulu, mention a compound 

isolated from the bulbs of Narcissus tazetta called pretazettine (Furusawa et al., 

1976a). This agent is one of many that have been investigated for their ability to


inhibit protein synthesis and prevent the growth of cancer cells and viruses 

(see Figure 13.1). 

During antiviral screening of medicinal plants from the Pacific area, Furasawa 

and co-workers found that the alkaloid pretazettine was active against Rauscher 

viral leukaemia and spontaneous T-cell leukaemia in mice (Furusawa et al., 1976b, 

1979). Pretazettine was inhibitory towards murine retroviruses in cell cultures and 

in vivo. The compound was shown to inhibit viral reverse transcriptase (Furasawa 

et al., 1978), and was cytotoxic due to a specific inhibitory action on cellular 

protein rather than nucleic acid synthesis and therefore active against non-viral 

transplantable tumours. Later studies established that pretazettine was active 

against Ehrlich ascites carcinoma – a non-viral, transplantable tumour – in mice 

(Furusawa et al., 1981) and that the compound enhanced the antitumour activity 

of a standard combination of adriamycin, 1,3-bis-(2-chloroethyl)-1-nitrosourea 

(BCNU) and cyclophosphamide – the so-called ABC regimen – when used as an 

adjuvant to the latter. Significant survival rates were seen in inoculated animals. 

The authors also demonstrated that pretazettine could inhibit protein synthesis, 

but not nucleic acid synthesis in human carcinoma cells in vitro. A further interesting 

finding was that adriamycin pre-treated, human carcinoma cells were more 

sensitive to pretazettine; therefore the drug could be used at lower doses which 

were not inhibitory when pretazettine was used on its own. This suggested to 

the authors that the compound might be useful as adjuvant therapy, eradicating 

residual tumour cells by preventing the synthesis of proteins, notably DNA polymerase 

beta, necessary for DNA repair. 

In addition to the above, preliminary experiments suggested that pretazettine 

was effective against intraperitoneally inoculated Lewis-lung carcinoma – another 

non-viral, transplantable tumour – in mice (Furusawa and Furusawa, 1983). Pretazettine 

was also studied when the inoculation was subcutaneous in singeneic 

mice, more resistant to chemotherapy (Furusawa and Furusawa, 1986). In the 

latter model, pretazettine was inhibitory to pulmonary metastases but not growth 

of the primary tumour or in prolonging life span. The compound was more effective 

against the primary tumour when combined with standard cytotoxic agents 

such as adriamycin, cisplatin, 5-fluorouracil, methotrexate and vincristine; these 

were not effective when administered individually. This indicates that pretazettine 

may be useful as an adjuvant to induce positive effects from otherwise inactive 

agents. In allogenic mice, pretazettine inhibited metastases and also prolonged life 

span. 

The same group has also investigated the activity of pretazettine against 

spontaneous T-cell leukaemia in mice compared with that of some established 

antileukaemic agents. The activity of pretazettine was shown to be superior to 

methotrexate, 6-thioguanine and adriamycin, but inferior to vincristine. Combination 

with vincristine, 6-thioguanine and adriamycin was synergistic whereas, 

combination with methotrexate was not. Pretazettine was also shown to reverse the 

leukaemia enhancing effect of cyclosporine in this animal model at the preleukaemic 

stage, although the mechanism was unclear (Furusawa and Furusawa, 

1988). The implications of this work for the treatment of similar cancers in man 

await confirmation by independent study. 

Pretazettine has been shown to be active against selected RNA-containing 

flaviviruses (Japanese encephalitis, yellow fever and dengue) and bunyaviruses

350 D. Brown 

(Punto Toro and Rift Valley fever) in organ culture. Activity in an animal model 

was not studied (Gabrielsen et al., 1992). This activity may reflect a general ability 

to inhibit protein synthesis during viral replication. 

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14 Galanthamine: clinical trials 

in Alzheimer’s disease 

David Brown 

INTRODUCTION 

The isolation, and subsequent chemical, pharmacological and toxicological characterisation 

of galanthamine, is described in the previous chapter of this volume, 

together with a summary of trials in indications other than Alzheimer’s Disease 

(AD). This chapter reviews the early trials of galanthamine in AD and provides 

detail, where it is available, of ongoing clinical research with the compound. For a 

review of the animal work underpinning this application, the reader is referred to 

the previous chapter. 

The development of galanthamine has been slow and clinically multi-faceted, 

but it is far from a dinosaur drug. At the time of writing, at least three pharmaceutical 

companies are developing the drug for the AD market on an international 

basis. Waldheim Pharmazeutika (Austria) has developed a method of synthesis 

that is feasible on an industrial scale. Shire Pharmaceuticals (England) has filed 

patents and is conducting extensive clinical trials with an allied company, Janssen 

Pharmaceutica (Belgium). The understandable reluctance of these companies to 

reveal unpublished data for inclusion in this review is evidence of the highly 

competitive atmosphere surrounding galanthamine. 

BRIEF OVERVIEW OF ALZHEIMER’S DISEASE 

AD is the most common form of dementia, accounting for 50–70% of all cases 

(Rossor, 1996) and is an important cause of morbidity and premature death in the 

elderly, worldwide. Typically, patients experience a slow but inexorable decline in 

memory and cognitive function, which eventually leads to complete dependency 

on family and professional carers and death, on average 4–6 years from diagnosis 

with a spread of 2–10 years. Early symptoms include short-term memory loss, 

progressing to confusion and disorientation with intermittent periods of lucidity, 

which often make the disease more painful to bear for patients and carers alike. As 

AD progresses, gross personality changes and emotional disintegration may occur. 

In long-standing disease, the patient may be mute, inattentive and completely 

incapable of self-care. Late psychiatric symptoms include hallucinations, agitation 

and aggression. Death commonly results from the complications of immobility 

such as bronchopneumonia; a definitive diagnosis of AD requires post-mortem

356 D. Brown 

examination of a brain biopsy, although clinical diagnoses are frequently used to 

decide upon management. 

It is estimated that over half a million people in the UK, 1.5 million in Japan 

and 4 million in the USA have AD (Office of Population Census and Surveys, 

1989). This burden is likely to increase as the average age of these populations 

increases (Jorm et al., 1991). The disease already places huge demands on health 

and social service budgets: for example, the UK spends in excess of £1000 

million (Gray and Fenn, 1993) and the USA some 50 times this amount (Office of 

Technology Assessment, 1987). It is not surprising, then, that so much effort has 

been put into research to understand AD and to find effective therapies. Comprehension 

of this complex disease is far from complete and, presently, realistic treatment 

objectives are to minimise either the degree or rate of cognitive decline and 

preserve functional ability. 

ANTICHOLINESTERASES IN ALZHEIMER’S DISEASE 

AD is characterizsed by depletion of a number of brain neurotransmitters, including 

acetylcholine (Mucke, 1997; Davies, 1983). By the late 1960s, the role of acetylcholine 

in the formation and maintenance of memory was firmly established. 

Seminal experiments demonstrated that a transient AD-like syndrome could be 

produced in laboratory animals and man with the centrally penetrating agent, 

scopolamine (Gruber et al., 1967; Crow and Grove-White, 1973). Furthermore, 

AD patients were shown to be more sensitive to cholinergic blockade than control 

individuals (Sunderland et al., 1987). Davies and Maloney (1976) were the first to 

report the selective loss of cholinergic neurones in AD patients and this finding 

was soon corroborated (Perry et al., 1978). Workers were able to localise the cholinergic 

deficit to those brain areas associated with cortical activation, notably the 

nucleus basalis, an important centre for cortical enervation (Whitehouse et al., 

1981). This deficit is common to the vast majority of AD patients studied. Bartus 

et al. (1982) finally advanced the cholinergic hypothesis of AD, based on accumulated 

experimental and clinical evidence. In this hypothesis, the primary defect is 

dysfunctional synthesis and secretion of acetylcholine from the pre-synapse, rather 

than alteration in catabolism by cholinesterase in the synaptic cleft (see Figure 

14.1). Consequently, intra-synaptic levels of acetylcholine are reduced, resulting in 

impairment of signal transmission to cortical regions. While acknowledging the 

multifactorial nature of the biochemical disorders in brain chemistry in AD 

patients (levels of noradrenaline, dopamine, serotonin, gamma-amino butyric 

acid, glutamate, somatostatin and substance P may also be reduced) and the likely 

role of amyloid protein plaque development in its pathophysiology, the cholinergic 

hypothesis remains a strong one on which to base intervention strategies to 

slow the mental deterioration associated with the disease (Unni, 1998). The 

abundance of experimental and clinical data, which forms this basis, means that of 

all the possible mechanisms, it is the best understood. Secondary benefits which 

might result from cholinergic restoration include an increase in local blood flow, 

thus easing vascular dementia (Geaney et al., 1990) and promotion of normal 

processing of the amyloid precursor proteins, thus reducing amyloid plaque 

formation (Giacobini et al., 1996).

Pre-synapse 

 

 

 

Synaptic cleft 

Clinical trials of galanthamine 357 

Post-synapse 

Figure 14.1 Schematic representation of the action of galanthamine. 

➀ Acetylcholine is synthesised and released by pre-synaptic tissue and diffuses across 

the synaptic cleft ➁. 

➂ In health, sufficient acetylcholine reaches post-synaptic tissue, where it interacts 

with receptors to ensure effective nerve transmission. In Alzheimer’s disease, according 

to the cholinergic hypothesis, acetylcholine levels are reduced and transmission is 

less effective. 

➃ Receptor binding is reversible; released acetylcholine diffuses back into the synaptic 

cleft and is degraded by pre-synaptic acetylcholinesterase ➄, producing choline ➅, 

which is recycled to produce more acetylcholine. 

Galanthamine binds reversibly with acetylcholinesterase, inhibiting the catabolism of 

acetylcholine, which is then free to replenish and sustain more effective levels in the 

synaptic cleft ➆. 

Based on this hypothesis, the aim should be to restore intrasynaptic levels of 

acetylcholine in AD patients to those seen in health. Various ways of doing this are 

reviewed elsewhere (Mucke, 1997; Bartus et al., 1985; Coyle et al., 1983). The 

method which holds most promise and therefore has found most favour is to prevent 

catabolism of existing intrasynaptic acetlycholine by inhibiting the enzyme 

responsible – acetylcholinesterase. A number of anticholinesterase drugs have 

been investigated: physostigmine (plus two longer acting derivatives, heptylphysostigmine 

and phenserine), tetrahydroaminoacridine (tacrine), velnacrine, metrifonate, 

donepezil and rivastigmine, the last two having been licensed in the UK for 

AD in 1997 and 1998, respectively. None of them, including galanthamine, is 

ideal. For example, tacrine, a well-researched drug with which galanthamine is 

often compared, has met with a modicum of success, but the frequency of side 

effects, notably hepatotoxicity (Davies and Powchik, 1995), restricts its use and 

mandates special patient monitoring. 

Early trials with galanthamine were at best inconclusive and of an anecdotal 

nature, but several centres are now working with galanthamine in thoroughly 

designed clinical trials, which should give us a clearer idea of the therapeutic 

potential of the drug in this distressing disease.

358 D. Brown 

A successful cholinergic therapy for AD would have to have the following 

properties: 

• Ability to cross the blood-brain barrier after oral or parenteral administration. 

• Adequate and predictable absorbtion after oral administration. 

• Unaffected by first-pass metabolism. 

• High affinity for brain acetylcholinesterase. 

• Ready reversibility of binding to brain acetylcholinesterase. 

• Selectivity for acetylcholinesterase rather than butyrylcholinesterase (present 

mainly outside the central nervous system). 

• A pharmacokinetic profile allowing once or twice daily dosing. 

• Few side effects, and those that do occur should be well characterised and 

reversible on drug cessation. 

Clinical studies to date indicate that galanthamine fulfils at least some of the 

criteria on this wish-list. 

CLINICAL TRIALS WITH GALANTHAMINE 

Early clinical trials 

Galanthamine was evaluated in the treatment of patients with AD in a few small 

trials that were essentially pilot studies. Most were non-comparative, open-label, 

with low patient numbers making statistical interpretation difficult or impossible. 

The results are often available only in abstract form and interesting data, such as 

subjective impressions, side effects and withdrawals, are often omitted. Furthermore, 

the diagnostic tests used to assess response to therapy have differed from 

study to study, making objective comparisons difficult. Table 14.1 contains definitions 

of the main tests used in galanthamine trials. Table 14.2 summarises the 

main points of galanthamine trials reported so far. Galanthamine is commonly 

administered as the hydrobromide salt. 

The first study to be published came from Austria (Rainer et al., 1989). The trial 

involved ten patients with symptoms indicating Alzheimer-type dementia. Nine 

remained evaluable, eight females and one male; the reason for withdrawal of one 

patient was not given. The age range of the nine evaluable patients was 61–89 

years (mean, 77 years). The mean duration of disease was 2.7 years. Patients were 

initiated on 15 mg galanthamine hydrobromide orally, increasing to 30 mg/day 

after 1 week and continuing for 7 weeks. While the pre- to post-treatment comparison 

did not reach statistical significance in most psychometric and neuropsychiatric 

tests, some improvements in clinical symptoms were observed. Patients 

showed improvement in some standard psychological tests of cognitive function 

and memory conventionally used on AD patients, notably tests of global memory 

performance, attention and concentration (six of nine patients) and visual-motor 

shape perception, visual memory and attentive concentration (six of nine patients). 

There was an improvement in speed but not the degree of performance in some 

patients. Three patients experienced improved quality of life and day-to-day 

performance. No adverse effects were noted.


Table 14.1 Key instruments used to assess AD symptoms in trials with galanthamine 

Instrument (reference) Description 

MMSE (Folstein et al., 1975) Mini Mental State Examination: standardised and 

validated test of cognitive function, covering memory, 

orientation, language and praxis. On a scale of 0–30, 

mild to moderate cognitive impairment is indicated by 

a score between 26 and 10. Less than 10 implies severe 

impairment. Score is influenced by age, socio-economic 

and educational status. 

ADAS-Cog (Rosen et al., 1984) Alzheimer’s Disease Assessment Scale – cognitive sub-scale: 

standardised scale, examining aspects of cognitive 

performance including memory, language and praxis. 

On a scale of 0–70, cognitive impairment is directly 

related to score. Typically, patients with mild to moderate 

disease progress by approximately 7–11 units per year. 

CIBIC-Plus (Jann, 1998) Clinician’s Interview-based Impression of Change scale: 

non-standardised scale assessing changes in patient 

performance in general, cognitive, behavioural and daily 

living activities; ranging from 1 – markedly improved, 

to 7 – markedly worse. 

Around the same time, a German group (Thomsen and Kewitz, 1989; Thomsen 

et al., 1990) reported on a 60-year-old female, suffering from advanced AD with a 

five-year history, who was given oral galanthamine hydrobromide in divided 

doses varying between 15 and 55 mg daily over a 120-day period. This included 

a 14-day washout period to examine the effects of drug withdrawal. Perhaps due 

to the advanced nature of her disease, the results of standard psychometric tests, 

including the widely accepted Mini Mental State Examination (MMSE) (Folstein 

et al., 1975; see Table 14.1), showed no appreciable improvement with therapy 

but neither was clinical deterioration observed over the extended period. Her 

doctor and spouse noted subjective improvement in the patient’s appearance 

and performance in daily tasks. Interestingly, cholinesterase inhibition in the 

patient’s erythrocytes was monitored throughout treatment, and these improvements 

corresponded with high-dose phases of therapy and a cholinesterase 

inhibition of 50–70%. Incidentally, the technique of titrating individual galanthamine 

doses to achieve a target level of erythrocyte cholinesterase inhibition 

was used in some, but not all, subsequent clinical trials. Galanthamine appeared 

to be well tolerated, without side effects except for transient tachycardia, excitation 

and headache that appeared upon re-introduction of the drug after the 

washout period. 

A second Austrian study (Dal-Bianco et al., 1991) examined 18 patients with 

the diagnosis of possible AD, made according to criteria defined by the National 

Institute of Neurological and Communicative Disorders and Stroke with the 

Alzheimer’s Disease and Related Disorders Association (NINCDS-ADRDA or 

simply NINCDS criteria; McKhann et al., 1984). Eighteen individuals, aged between 

53 and 83 years, were given galanthamine, 30–50 mg/day for periods ranging 

from 2–16 months. In this rather incompletely reported study, there were no

Table 14.2 Summary of human trials with galanthamine in Alzheimer’s disease 

Major outcomes Side effects Comments 

Design/tests used a Galanthamine daily 

dosage and duration 

Reference Country Patients (m = male, 

f = female) 

None reported Pilot study 

No statistically significant 

improvements in psychometric 

and neuropsychometric tests. 

Some individual improvements 

in memory, attention, concentra- 

15–30 mg, for 

7 weeks 

Open label, tests 

unspecified 

Austria 10, but 9 remaining 

evaluable, with 

AD-type dementia. 

8 f, 1 m, mean age 

77 years (range 

61–89) 

Germany 1f with advanced 

AD, aged 60 years 

Rainer 

et al., 

1989 

Pilot study. 

Most improvement 

noted with higher 

doses and greater 

erythrocyte 

cholinesterase 

inhibition. 

Effects reversed 

Transient tachycardia, 

excitation, 

headache 

tion and quality of life 

Case study. No significant change 

in MMSE; subjective improvement 

in patient appearance and 

performance of daily tasks 

15–55 mg, for 

120 days 

Open label, 

MMSE 

Thomsen 

and 

Kewitz, 

1989 

during washout 

None reported Pilot study. Effects 

of galanthamine 

reversed on 

withdrawal 

No statistically significant 

improvements. At 12 months, 

the 6 remaining patients showed 

positive changes in everyday 

function and emotional stability 

30–50 mg, for 

2–16 months. 10 

patients remaining 

at 6 months, 

6 patients remain- 

Open label. 

Tests 

unspecified- 

‘standard 

neurophysio- 

Austria 18 with ‘possible’ 

AD (NINCDS), 

aged 53–83 years 

Dal-Bianco 

et al., 

1991 

Pilot study. 

Deterioration 

noted during 

washout period. 

7 stable after 

9 months, 4 stable 

after 12 months 

7 withdrawals due to 

mainly gastro-intestinal 

side effects on high 

dose regimen 

Non-statistically significant gains 

in MMSE score. 

Significant improvements in 

CGIC (

No significant 

correlation 

between dose 

and cognitive 

improvement 

Nausea and vomiting 

dose-related: mean 

dose 29.4 mg: 21%; 34.7 

mg: 29%; 37.9 mg: 63%. 

Diarrhoea, 4%; abdominal 

cramps, 4%, weight 

loss, 1.2%, anorexia, 

3%. 78% tolerated individual 

best dose 

Mean improvement in ADAS- 

Cog of 5.14, MMSE 1.72 after 3 

weeks dose adjustment phase. At 

13 weeks, further improvement 

(1.7 points) in ADAS-Cog. 

Patients taking placebo deteriorated 

by 1.4 points. MMSE 

improved by 1.7 at 3 weeks and 

2.5 at 13 weeks; placebo score 

dropped 1.7 points below baseline. 

Changes in CGIC described 

Initial 3-week 

dose titration 

phase, then 

20–50 mg best 

dose or placebo 

for 10 weeks 

Multicentre. 

Randomised, 

double-blind, 

placebo 

controlled 

after initial 

recruitment 

of responders. 

MMSE 

ADAS-Cog 

CGIC 

141 with mild/ 

moderate, 

probable AD 

(NINCDS) 

International 

Berzewski 

et al., 

1994 

Pilot study. 

Number of patients 

on which analyses 

based unspecified 

Nausea and vomiting at 

start of therapy. Light 

headedness, agitation 

and sleep disturbance. 

Blood chemistry 

normal 

as favourable on galanthamine 

Mean improvement in ADAS- 

Cog of 1.33 with galanthamine 

and deterioration of 0.81 points 

with placebo. Results not statistically 

significant. Significant 

improvement in physicians global 

evaluation on galanthamine 

20–50 mg for 

10 weeks 

Multicentre. 

Double-blind, 

placebo 

controlled. 

ADAS-Cog 

Germany 95 (60–87 years) 

with mild/moderate 

primary degenerative 

dementia; 

recruited after a 

positive initial response 

under single 

blind conditions 

England 235 with mild/ 

moderate AD 

(NINCDS) 

Kewitz 

et al., 

1994 

Interim report of 

on-going trial. 

Some evidence of 

dose-response 

correlation. 

Authors suggest 

that 30 mg/day may 

be the optimum, to 

balance side effects 

with efficacy 

Incidence of nausea 

dose-related: 0, 13, 18, 

35% in placebo, 22.5, 

30.0 and 45.0 mg 

galanthamine groups. 

The groups had 8, 19, 

12 and 38% withdrawals 

respectively. Adverse 

events occurred mainly 

during dose escalation. 

No changes in liver 

In patients remaining at 12 

weeks, galanthamine attenuated 

decline in ADAS-Cog at all 3 

doses. In the 30 mg/day group, 

difference from placebo 

(3.4 points) statistically, highly 

significant, on intention 

to treat basis. Positive trends in 

CIBIC Plus also observed 

22.5, 30.0 or 

45.0 mg for 

10 weeks 


double-blind, 

placebo 

controlled 

after initial 

1/2 week dose 

adjustment. 

ADAS-Cog 

CIBIC Plus 

Wilcock 

and 

Wilkinson, 

1997 

function tests 

Not reported Pilot study. 

Improvements 

reversed on 

stopping drug. 

No statistical tests 

reported 

‘Significant’ improvement in 

attentional tasks starting as 

early as 2 weeks 

After 4-week dose 

titration, either 30 

or 40 mg for 12 

weeks 


double-blind, 

parallel group. 

Battery of 

attentional tests 

England 30 with AD 

(diagnostic 

criteria not 

reported) 

Wesnes 

et al., 

1998 

Notes 

a 

ADAS-Cog: Alzheimer’s Disease Assessment Scale – Cognitive Subscale (a positive number indicates decline and a negative number, improvement); CIBIC Plus: 

Clinician Interview Based Impression of Change; CGIC: Clinical Global Impression of Change; MMSE: Mini-Mental State Examination.

362 D. Brown 

statistically significant improvements in non-standard neuropsychological tests of 

verbal, non-verbal, language, motor or attention functions at 2 months, nor at 

6 months in ten patients still receiving the drug. At 12 months, six patients still 

taking galanthamine showed positive changes in everyday function and/or emotional 

stability, as noted by their carers. These patients appeared to deteriorate during 

drug withdrawal periods of unspecified length. No adverse events were reported. 

This pilot study suggested that there may be a sub-group of AD patients responding 

to galanthamine, whose pathology had a major component of cholinergic deficit. 

In an effort to determine any relationship between dose and clinical benefit, 

Wilcock et al. (1993) treated 19 probable AD patients (as defined by NINCDS 

criteria) with either low dose (30–40 mg) or high dose (45–60 mg) daily doses of 

galanthamine for two, six-week periods, separated by a three-week washout. Seven 

patients withdrew due to unacceptable side effects (mainly gastrointestinal, in the 

high-dose group) leaving 12 evaluable patients: five males with a mean age of 58 

years (range 48–74 years) and seven females of mean age 67 years (range 55–79 

years). Patients were assessed by a battery of recognised tests of neuropsychological 

and mental function, including the MMSE and the Alzheimer’s Disease Assessment 

Scale (ADAS, or if referring to the cognitive section alone, then ADAS-Cog, 

after Rosen et al. (1984); see Table 14.1). While a trend towards improvement was 

observed in several of these tests which, curiously, was most marked during the 

lower rather than higher dose regimen, the sole statistically significant change was 

an improvement in the cognitive component of the ADAS. Some deterioration was 

seen during the washout phase. A clinical, but statistically insignificant, improvement 

was recorded in the MMSE during the low-dose regimen; no clinically or 

statistically significant changes were observed in the Functional Life Scale, the 

Digit Span test or the Relatives’ Stress Scale. Four patients extended from the 

trial on galanthamine remained stable for 12 months; three others were stable for 

9months. 

These early pilot studies do provide an impression of benefit for galanthamine 

in AD; however, larger clinical trials were clearly needed to examine issues such as 

what dose to use in relation to disease severity, duration of therapy and medium to 

long-term side effect profile. 

Later clinical trials 

As with earlier work, interpretation of the results is made difficult by the 

nature of the publications. Most trials are reported in abstracts of symposium 

proceedings and, while the most important information is present, qualifying 

data on side effects, withdrawals, dose adjustments and assessment techniques 

are lacking. 

Rainer et al. (1994) recruited 58 patients with AD who were treated with galanthamine, 

20–50 mg daily for 3 weeks, during a single-blind preliminary investigation 

to find galanthamine responders for entry in a subsequent placebo-controlled 

trial. Galanthamine produced a mean decline (which represents a favourable 

effect) in the ADAS-Cog of 4.21 (from 32.72 to 28.51), while the MMSE score 

increased to 20.15 from 18.65 (a favourable result, indicating improvement in 

cognitive function). As with any such short-term open study, improvement might 

have been a practice, rather than a drug effect.


Berzewski et al. (1994) reported in abstract a multi-centre, international, 

placebo-controlled trial involving 141 patients with mild or moderate probable AD 

(according to NINCDS criteria) who were started on a 3-week, single blind dose 

titration phase of 20 mg galanthamine daily. Doses were titrated upwards in 10 mg 

steps, every 3 days until individual best-doses were reached, aiming for inhibition 

of erythrocyte cholinesterase by at least 40% of pre-therapy baseline. The upper 

dose limit was 50 mg/day. These patients showed a mean dose-related improvement 

of 5.14 ± 0.53 ADAS-Cog points, which is similar to that reported by Wilcock 

et al. (1993). Patients were then randomised to continue galanthamine at a personal 

best dose or switched to placebo for the next 10-week, double-blind phase. 

The results were encouraging. At the end of the study, those on galanthamine had 

improved by a further 1.66 ADAS-Cog points, but those on placebo had deteriorated 

by an average of 1.40 points. The MMSE scores showed a similar trend, with 

modest improvements during the optimisation phase (mean increase: 1.72 points) 

which rose to 2.50 points at the end of the trial. The placebo group dropped an 

average of 1.70 points below baseline. Standard neuropsychiatric tests reflecting 

drug effects in both cognitive and global evaluation scales showed the same 

favourable trend in 72% of patients treated with galanthamine – particularly the 

assessment of clinical global impression (effectiveness, tolerance and acceptance by 

the patient). Cholinergic side effects were mild, transient and dose-related. For 

example, the incidence of nausea and vomiting was 21, 29 and 63% in subgroups 

of patients taking average galanthamine doses of 29.4, 34.7 and 37.9 mg/day. 

Other cholinergic side effects were diarrhoea and abdominal cramps (4% each), 

weight loss (1.2%) and anorexia (3%). The study failed to demonstrate a statistically 

significant relationship between dose and cognitive improvement, although a 

trend was evident. 

Kewitz et al. (1994) reported a multi-centre, placebo-controlled, double-blind 

trial of the safety and efficacy of galanthamine in 95 patients, aged 60–87 years, 

with mild to moderate primary degenerative dementia. These patients had 

demonstrated a primary response to galanthamine under single blind conditions. 

Galanthamine was started at 10 mg twice a day and was increased to a 

maximum of 50 mg/day during the 3 weeks of dose optimisation. While there 

were no significant changes between performance on placebo and galanthamine 

in terms of performance on the ADAS-Cog scale (–0.81 for placebo, +1.33 for 

galanthamine), there was a significant improvement in the physicians’ global 

evaluation demonstrating significantly less deterioration in patients receiving 

galanthamine. Nausea and vomiting were the most common side effects, occurring 

usually at the start of therapy and tending to resolve with continued treatment. 

Light-headedness, agitation and sleep disturbances were also noted. No 

alterations in blood chemistry or liver function were noted. Some withdrawals 

due to gastrointestinal symptoms were noted, but these were not quantified in 

this abstract. 

Wilcock and Wilkinson (1997) reported the interim results of an on-going, 

randomised, double-blind study. After an initial adjustment phase lasting up to 14 

days, 235 patients with mild to moderate AD, as judged by the NINCDS criteria, 

received placebo or 22.5, 30.0 or 45.0 mg/day of galanthamine, in three divided 

doses with food. Therapeutic effects were measured by the ADAS-Cog scale and 

the CIBIC Plus scale (global Clinician Interview Based Impression of Change

364 D. Brown 

( Jann, 1998); see Table 14.1). Adverse events were recorded and haematological and 

biochemical parameters were monitored. Patients were assessed at baseline and at 

6 and 12 weeks during treatment. An analysis of patients remaining in the trial at 

12 weeks showed that the cognitive performance of placebo-treated patients had 

deteriorated during the treatment phase, but this was attenuated in all the 

galanthamine groups, with mean ADAS-Cog scores one or two points greater than 

zero. A trend in relationship to dose was apparent, although no statistical analysis 

of this was reported. The differences between the placebo and active scores for the 

30 mg and 45 mg/day groups were 3.8 and 3.9 points, respectively. Positive trends 

on the CIBIC Plus scale were also associated with galanthamine. On an intentionto-treat 

basis, the 30 mg/day dosage produced a statistically significant rise in 

ADAS-Cog score of 3.4 points from placebo. Side effects were mostly related to the 

cholinergic action of the drug, and occurred during the initial dose adjustment 

phase. The incidence of nausea was 0, 13, 18 and 35% in the placebo, 22.5, 30 and 

45 mg/day groups, respectively; the corresponding incidences for vomiting were 6, 

19, 7 and 17%, respectively. Overall, galanthamine was described as being well 

tolerated, although 38% of patients in the 45 mg/day regimen (compared with 8, 

19 and 12% of patients taking placebo, 22.5 mg or 30 mg doses, respectively) withdrew 

due to cholinergic side effects, mainly during the dose-escalation phase. This 

is an indication of the likely maximum dose at which galanthamine may be tolerated 

or, at least, a warning that if such high doses are to be used, a more cautious 

dose escalation technique should be employed. No abnormalities in liver function 

tests were observed. The authors concluded that patients received optimal benefit 

from 30 mg/day, and that a longer titration period to achieve this dosage might 

avoid the cholinergic symptoms of more rapid initiation. 

Wesnes et al. (1998) published a brief report focusing on the effect that galanthamine 

might have on the attention deficit seen in AD patients. They described a 

double-blind, parallel group trial in 30 patients randomised to receive either 30 or 

40 mg/day of galanthamine for 12 weeks, following a dose titration period of 4 

weeks. Attention deficit was measured using an attentional sub-battery of tests of 

cognition. No statistical analysis was reported, but improvements described as 

significant were observed in all three attention tests used, starting early on in 

therapy, increasing with time, but which rapidly declined on stopping galanthamine. 

The attention tests used in this trial were not part of the more usual 

ADAS-Cog test battery, and the authors pointed out that the tests they used may 

provide important additional information on the use of drugs for AD. 

Longitudinal studies 

Patients with AH may live for many years and clearly, the long-term efficacy and 

safety of a therapeutic agent used in the disease is of great importance. As yet, few 

data are available for galanthamine. Recently, the interim results from an open, 

3-year follow-up evaluation of patients using galanthamine have been published 

(Rainer and Mucke, 1996). Patients receiving galanthamine with or without additional 

non-anticholineterase therapy (data reported on 21), such as antidepressants 

and nootropics, showed a significantly lower rate of cognitive decline than 

those taking placebo (data reported on 23), as shown by the mean ADAS-Cog 

scores. The degree of cognitive stabilisation achieved during the first year was a


good indicator of 3-year outcome. No clinically significant disturbances in blood 

biochemistry or liver function tests were observed. 

SUMMARY 

The results of early trials of galanthamine compare well with those of later studies, 

but one should remember that today’s criteria for classification of patients with 

AD are much more precise. It has been estimated that early inclusion criteria may 

have been only 85% specific for AD (Wade et al., 1987). Most of the early studies 

reported here are really no more than pilots, and leave much to be desired; however, 

they were conducted by three, independent research groups and the results attracted 

sufficient attention to prime the planning of more extensive and sophisticated trials. 

Such trials are at present ongoing and results have not yet been published. 

One reason why the anticholinesterase approach is only partially successful in 

AD is the multifactorial nature of the neuropharmacological deficits seen in the 

disease (Perry, 1987). As AD appears to be a disorder involving deficits in a 

number of neurotransmitters, it seems logical to assume, and on the basis of the 

evidence, correct, that the strategy of acetylcholine preservation slows the progression 

of the disease but does not halt it. Secondly, a multifactorial approach, 

involving several pharamcological interventions at once (for example, combined 

strategies to boost levels of noradrenaline, serotonin, gamma-aminobutyric acid, 

somatostain and corticotrophin releasing factor, all of which have been shown to 

be deficient in AD), may hold more promise. 

From the trial data so far, it would appear that there is a sub-group of patients 

who respond well to galanthamine, and in whom the cholinergic deficit is a major 

component of their disease. 

The side effect profile of galanthamine appears favourable, particularly when 

compared with tacrine, where hepatotoxicity is a particular worry (Knapp et al., 

1994; Wagstaff and McTavish, 1994) and metrifonate, where clinical trials have 

recently been suspended because of concerns about muscle weakness (personal 

communication, Bayer UK). Anticholinergic side effects, particularly nausea and 

vomiting, may be a problem, and measures to minimise these, such as slow 

upward titration of the initial dose and co-administration of antiemetics, should be 

investigated further. 

Galanthamine is approved by the Austrian authorities for use in AD. However, 

there remains at present a lack of controlled, double-blind trials of sufficient quality 

and length to define clearly the role of this interesting agent. Head-to-head 

comparisons with other agents such as tacrine have not been performed, and 

more information is also required on long-term use, dose response, use in old age 

(where renal and hepatic impairment are common), and potential interactions 

with other drugs used in co-morbid conditions (such as Parkinson’s disease, agitation, 

anxiety, aggression and depression). 

In terms of pharmaceutical development, a once-daily dosage form would be a 

useful advance, having obvious applications in elderly or forgetful patients and at 

least one transdermal, controlled release patch is under development (Morierty, 

1995). An alternative is to synthesise galanthamine analogues with extended halflife 

and improved efficacy.

366 D. Brown 

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15 Screening of Amaryllidaceae 

for biological activities: 

acetylcholinesterase inhibitors 

in Narcissus 

Kornkanok Ingkaninan, Hubertus Irth and Rob Verpoorte 

INTRODUCTION 

Narcissus are well-known cultivated plants belonging to the Amaryllidaceae family. 

Tyler et al. (1988) classified them as poisonous plants, since ingestion of narcissus 

bulbs produces severe gastroenteritis and nervous symptoms. Narcissus plants 

contain similar alkaloids which can also be found in other members of the Amaryllidaceae. 

The Amaryllidaceae alkaloids have been shown to possess a wide spectrum 

of biological activities such as anticholinergic and analgesic (Harvey, 1995), 

antimalarial (Likhitwitayawuid et al., 1993), antiviral (Gabrielsen et al., 1992; Lewis, 

1996) and anti-neoplastic (Jimenez et al., 1976; Ghosal et al., 1988; Pettit et al., 

1990, 1993). However, galanthamine is so far the only Amaryllidaceae alkaloid 

that has gained a wide spread commercial application because of its inhibitory 

activity for acetylcholinesterase (AChE). 

GALANTHAMINE 

Galanthamine is a tertiary amine alkaloid belonging to the phenanthrene group. 

It was first isolated from bulbs of Caucasian snowdrop, Galanthus woronowi, in 1952 

(Proskurnina and Yakovleva, 1952). It has also been found in various species of 

Narcissus (Paskov, 1986; Bastida et al., 1990). The compound was first applied for 

medical purpose by Bulgarian and Russian researchers in the 1950s. In 1955, 

Mashkovskii reported that galanthamine could reverse tubocurarine-induced 

muscle paralysis and potentiate the actions of acetylcholine on skeletal muscles. It 

was then used as a potent natural cholinergic substance showing strong AChE 

inhibitory activity in both the central and peripheral system, and for the reversal 

of the neuromuscular blockade caused by various curare-like agents (Irwin and 

Smith, 1960; Cozanitis, 1971; Baraka and Cozanitis, 1973; Cozanitis and Rosenberg, 

1974; Krivoi, 1988). This drug has been available for clinical use as Nivalin ® 

(Pharmachim, Sofia, Bulgaria) and Galanthamine ® (Medexport, Moscow, USSR). 

It was not until 1986, after the cholinergic hypothesis of Alzheimer’s disease was

370 K. Ingkaninan, H. Irth and R. Verpoorte 

postulated, that the application of galanthamine in Alzheimer’s disease was 

studied. Based on the cholinergic hypothesis (Perry, 1986), memory impairments 

in patients suffering from this disease result from a defect in the cholinergic system. 

One approach to the treatment for this disease is to enhance the acetylcholine 

level in the brain by AChE inhibitors (Winblad et al., 1993). Galanthamine hydrobromide 

is now being developed for Alzheimer’s disease. It has shown clear evidence 

for being suitable for the treatment of mild and moderate Alzheimer’s disease. At 

least three corporate pharmaceutical companies are involved in developing galanthamine 

for the Alzheimer’s disease market: Waldheim Pharmazeutika (Austria) 

for developing a method of synthesis, Shire Pharmaceuticals (UK) for clinical studies, 

and Janssen Pharmaceutica (Belgium) for final registration and marketing. 

The preclinical and clinical studies of galanthamine were reviewed by Mucke 

(1997a,b) and Rainer (1997), respectively. 

Galanthamine is commercially isolated from Leucojum aestivum (Amaryllidaceae) 

(Paskov, 1986). Although the chemical synthesis of galanthamine was accomplished 

in 1960 (Barton and Kirby, 1960), it was only very recently that an industrial 

scale synthesis of galanthamine hydrobromide was established (Czollner et al., 

1996). Several analytical techniques have been described to quantify galanthamine 

in natural sources. Kreh et al. (1995) and Bastos et al. (1996) reported capillary 

column gas chromatographic methods for quantification and identification of 

galanthamine and other Amaryllidaceae alkaloids. A radioimmunoassay developed 

by Tanahashi et al. (1990) provided specific and precise quantitation of galanthamine 

in unpurified plant extracts. Three years later, Poulev et al. (1993) established a 

more sensitive enzyme immunoassay for the quantitation of fmol amounts of 

galanthamine. 

SCREENING ASSAYS FOR ACETYLCHOLINESTERASE INHIBITORS 

Several assays have been established for the detection of AChE activity and its 

inhibitors. The colorimetric method developed by Ellman et al. (1961) is the most 

widely used. This assay is based on the enzymatic hydrolysis of acetylthiocholine 

iodide (ATCI) to yield thiocholine. When thiocholine reacts with 5,5′-dithiobis-2nitrobenzoate 

(DTNB), it will produce the yellow product of 5-thio-2-nitrobenzoic 

acid which can be detected at 405 nm by a spectrometer. 

acetylthiocholine + H2O AChE 

acetate + thiocholine 

thiocholine + DTNB 5-thio-2-nitrobenzoic acid 

+ 2-nitrobenzoate-5-mercaptothiocholine 

This spectrometric assay has been used for the screening of plant extracts for anti- 

AChE activity (Park et al., 1996; Kim et al., 1999). The use of a 96-well plate reader 

for measuring the AChE activity, reported by Ashour et al. (1987), allowed the 

rapid screening of series of samples. 

A radiometric technique for the detection of AChE activity based on hydrolysis 

of [ 3 H] labelled acetylcholine was reported by Johnson and Russell (1975). Guilarte 

et al. (1983) developed a simpler method using [ 14 C]sodium bicarbonate. The 

acetic acid formed from the enzymatic hydrolysis of acetylcholine reacts with

Acetylcholinesterase inhibitors 371 

[ 14C]sodium bicarbonate to generate [ 14C]carbon dioxide, which is measured using 

an ionisation chamber system. Compared with Ellman’s colorimetric method, these 

radiometric techniques are more sensitive. However, the radiometric method is an 

endpoint measurement and cannot be used for kinetic studies. Furthermore, the 

limited linear range of the method demands the proper calibration of the activity 

prior to incubation. 

A fluorometric method has also been developed based on the reaction of 

thiocholine produced by the enzyme from acetylthiocholine with the fluorogenic 

compound N-(4-(7-diethylamino-4-methylcoumarin-3-yl)phenyl) maleimide (CPM) 

(Parvari et al., 1983). This assay combines the specificity and the technical advantages 

of the Ellman technique with a wide linear range of accuracy and a limit of 

detection which is 100-fold lower than that of the radiometric method. Roger et al. 

(1991) developed the optical sensor for anti-AChE using immobilised fluorescein 

isothiocyanate (FITC)-tagged eel electric organ AChE on quartz fibres for monitoring 

enzyme activity. This biosensor is reusable, sensitive and easy to operate. 

It shows potential adaptability to field use as the instrument is portable. 

Recently, Pasini et al. (1998) reported a chemiluminescent (CL) method for a 

384-well microtiter format assay for high throughput screening of AChE inhibitors. 

The CL detection of AChE was based on coupled enzymatic reactions involving 

choline oxidase and horseradish peroxidase as the indicator enzymes. The reaction 

leads to light emission and luminol is used for the detection of the peroxide 

formed. This method had a detection limit 1000-fold lower than that of the colorimetric 

method. 

THE SEARCH FOR AChE INHIBITORS FROM NATURAL SOURCES 

Nature is a rich source of biological and chemical diversity. It has been shown 

many times that natural products could be developed as drug candidates. The 

unique and complex structures of natural products such as paclitaxel cannot be 

obtained easily by chemical synthesis. The well-known AChE inhibitors, galanthamine 

and physostigmine, are also obtained from plants. The clinical studies of 

AChE inhibitors for Alzheimer’s disease are still ongoing. Until now, no drug of 

choice has been decided upon. Therefore, the search for new AChE inhibitors 

from natural sources is of great interest. However, in searching for new leads from 

crude natural products extracts, one might encounter the problem of isolation of 

known active compounds or compounds that give aspecific effects on the bioassays 

used. A strategy of ‘dereplication’ of active compounds, or rapid identification of 

known active compounds at an early stage, is important in the lead finding 

process. One of the approaches for dereplication is to couple a separation technique 

with a bioactivity detection method. The known active compounds can be 

rapidly identified by the aid of data analysis and the unknown active fraction can 

be further investigated. 

Narcissus plants are a potential source for AChE inhibitors as they contain a 

variety of compounds, especially alkaloids, the group of compounds that shows 

many biological activities including AChE inhibitory effect. Since narcissus are 

cultivated plants, there could be a large variation of chemical constituents among 

different cultivars. Figure 15.1 shows the inhibitory effect of the methanol extracts


% Inhibition of acetylcholinesterase 

100 

90 

80 

70 

60 

50 

40 

30 

20 

10 

0 

Avalanche Carlton Grand 

Soleil d'Or 

Sir Winston 

Churchill 

Figure 15.1 Percentage of inhibition effect of methanol extracts from some narcissus 

cultivars at the concentration of 0.1 mg/ml measured with the microplate 

assay. 

from some narcissus cultivars tested in our laboratory. The microtiter plate assay 

used was according to the method of Ellman et al. (1961), as described by Ingkaninan 

et al. (2000a). All extracts showed an inhibitory activity for AChE. However, the 

activity might be due to the known active compound, galanthamine. A dereplication 

step for the rapid identification of galanthamine in crude extracts is necessary. 

The extracts also contain other unknown, active compounds that will be selected 

for further studies. 

IDENTIFICATION OF AChE INHIBITORS BY HPLC WITH ON-LINE 

COUPLED UV, MS AND BIOCHEMICAL DETECTION 

Recently, strategies for the on-line coupling of biochemical detection to separation 

methods such as high-performance liquid chromatography (HPLC) have been 

developed (Irth et al., 1995). This technique allows the simultaneous separation 

and detection of bioactive compounds from complex mixtures such as natural 

products. To obtain additional information on the compounds separated, the 

on-line system can also be coupled with other detection methods such as mass 

spectrometry (MS) and ultraviolet (UV) or photodiode array (PDA) detection. In a 

natural products-based drug discovery programme, this technique can be useful 

for the detection of new active compounds in the presence of the already known 

active compounds. It opens the possibility for the rapid screening of galanthaminecontaining 

plants for other AChE inhibitors. 

The colorimetric assay for AChE as reported by Ellman et al. (1961) is suitable 

for development into a continuous-flow biochemical detection system. AChE from 

electric eels is inexpensive and has satisfactory stability at room temperature. The 

assay reagents, DTNB and ATCI, are also commercially available. The short reaction 

time of the assay, approximately 2 min, made it possible to develop it for continuousflow 

biochemical detection system without causing much band-broadening.

Identification section 

Separation section 

HPLC 

pump 

injector 

Bioactivity test section 

DTNB 

pump 

ATCI 

pump 

HPLC column 


50°C 

AChE 

pump 

Figure 15.2 shows the scheme of the on-line HPLC-UV-MS-biochemical detection 

system developed in our laboratory (Ingkaninan et al., 2000a). The separation 

is performed in an HPLC column. The eluate is split into three flows: the major 

flow travels to a UV detector, while two minor flows go to the MS and the 

biochemical detection system. In the biochemical detection system, the eluate is 

mixed with substrate (ATCI), enzyme (AChE) and DTNB for approximately 2 min 

in the reaction coil. A spectrometer set at 405 nm detects the yellow product 

obtained from the enzymatic reaction. Any inhibitory activity from the HPLC eluate 

will result in a negative peak on the biochemical detector. After determination of 

the delay times of the three detection lines, the results of these detections can be 

related and thus UV spectra and molecular weight of the active compounds can 

be determined. In this way, known inhibitors such as galanthamine can easily be 

recognised. 

The strongly AChE inhibiting extract from narcissus ‘Carlton’ (Figure 15.1) was 

fractionated by means of centrifugal partition chromatography (CPC) and the 

active fraction was injected into the on-line HPLC-UV-MS-biochemical detection 

system (Ingkaninan et al., 2000a). The chromatogram from two detectors and the 

mass spectra are shown in Figure 15.3. Although the chromatogram from the 

HPLC is very complex, the biochemical detection system showed a high selectivity 

for the AChE inhibitors. Two broad negative peaks from the biochemical detector 

proved the presence of at least two inhibitors in the extract. The retention time of 

the main active peak and the molecular weight derived from MS data ([M + H] 

of 288) corresponded to those of galanthamine, a well-known AChE inhibitor. 

MS 

UV or PDA 

Detector 

Vis detector at 

405nm 

Figure 15.2 Scheme of the on-line HPLC-UV-MS-bioactivity detection for acetylcholinesterase 

inhibitors.


Figure 15.3 The spectra and the chromatograms obtained after injection of the fraction 

from narcissus ‘Carlton’ extract into the on-line HPLC-UV-MS-bioactivity 

detection system. Left: ESI-MS. Right: Chromatograms from the biochemical 

detection set at 405 nm (upper line) and from the UV set at 215 nm 

(lower line). The delay time of the peaks between the biochemical detection 

and the UV was 2.3 ± 0.1 min and the delay time of the peaks between 

the biochemical detector and the MS was 2.1 ± 0.1 min. 

However, MS showed that there was another molecule present in the same peak at 

[M + H] of 290. It would be interesting to identify this compound further and test 

it for inhibitory effect. The minor peak in the chromatogram detected by the biochemical 

detector was caused by an unknown AChE inhibitor. Further studies 

should, therefore, focus on the isolation and identification of this active compound. 

A second example is a study of the extract from narcissus ‘Sir Winston Churchill’ 

(Ingkaninan et al., 2000b). From the microplate assay, this extract showed much 

less activity than that of narcissus ‘Carlton’ (Figure 15.1). The extract was fractionated 

by the same CPC procedure as that of narcissus ‘Carlton’. The active fraction 

was also obtained at the same retention. However, when the active fraction was 

injected into the on-line system, no activity corresponding to galanthamine was 

found. The activity was from another compound with a different molecular weight 

and retention time. As this extract possibly contains new AChE inhibitors, it was 

chosen for further investigation. The active fraction was further separated by 

another CPC run and the fractions obtained were injected into the on-line system 

(Figure 15.4). 

The active peak from the biochemical detector at 15.2 min corresponded to the 

UV peak at 12.9 min and the MS peak at 13.1 min. However, MS spectra showed 

that there were two compounds present at 13.1 min having molecular weights of 

265 ([M + H] of 266) and 317 ([M + H] of 318). These two compounds were 

isolated by preparative HPLC and tested for AChE inhibitory activity by the


Figure 15.4 The spectra and the chromatograms obtained after injection of the fraction 

from narcissus ‘Sir Winston Churchill’ extract into the on-line HPLC- 

UV-MS-bioactivity detection system. Left: ESI-MS. Right: Chromatograms 

from the biochemical detection set at 405 nm (upper line) and from the 

UV set at 215 nm (lower line). The delay time of the peaks between the 

biochemical detection and the UV was 2.3 ± 0.1 min and the delay time of 

the peaks between the biochemical detector and the MS was 2.1 ± 0.1 min. 

O 

O 

HO 

H 

16 

OCH3 

OH 

3 

H 

N 

HCO 

3 

ungiminorine galanthamine 

O 

microplate assay. The inhibitory activity was found only in the compound with 

molecular weight of 265. This active compound was identified as an alkaloid, 

ungiminorine (Figure 15.5). This compound was first isolated by Normatov et al. 

(1965). The IC 50 value of ungiminorine was 86 ± 7 µM while that of galanthamine 

was 0.98 ± 0.07 µM. Although this compound shows much lower AChE inhibitory 

activity compared with that of galanthamine, it clearly demonstrates the usefulness 

of the on-line HPLC-UV-MS-biochemical detection for the search of new leads 

from natural products. 

N 

CH3 

OH 

Figure 15.5 Structure of ungiminorine and galanthamine.


FUTURE PERSPECTIVES 

It is to be noted that in the on-line HPLC-UV-MS-biochemical detection, only a 

small amount, i.e., one-thirtieth of the eluate from the HPLC, was split to the 

biochemical detector. Therefore, it is possible to apply the on-line system for a 

preparative separation technique such as preparative HPLC or CPC and on-line 

fraction collection can be performed. In both separation techniques, gradient 

elution might be useful for broad range separation. Furthermore, to gain more 

information for the identification of active peaks, a PDA and an NMR can be coupled 

to this on-line system. 

Although the Ellman colorimetric method used in this on-line biochemical 

detection system is not the most sensitive, it is the simplest and the most 

inexpensive screening method for AChE inhibitors. In natural productsbased 

research, where milligrams of new leads are still needed for the structure 

elucidation, selectivity for the bioactive compounds is more important 

than sensitivity. However, the sensitivity of the on-line biochemical detection 

could be increased by using a fluorometric method. Some of the fluorescent 

probes such as N-[4-[7-(diethylamino)-4-metylcoumarin-3-yl]phenyl]maleimide 

(CPM), which forms a fluorescent product when it reacts with thiocholine 

(Berman and Leonard, 1990) or the substrate, 7-acetoxy-N-metylquinolinium 

iodide (Menger and Johnston, 1991), have been used in an ultrasensitive 

assay for acetylcholinesterase and are commercially available. These fluorescent 

probes could be further developed for the on-line biochemical detection 

system. 

Another method that could be useful for the screening of AChE inhibitors from 

natural products is thin-layer chromatography (TLC). Some TLC methods using 

an enzyme, a substrate and a dye as the spraying reagent have been reported 

(Bunyan, 1964; Menn and McBain, 1966; Kiely et al., 1991). The inhibition zone 

from the TLC indicates the inhibitory activity of the test compound to the enzyme. 

When complex mixtures such as crude plant extracts are tested, the presence of 

known inhibitors in the extract will be rapidly identified. This technique can be 

helpful in the selection of extracts to be further investigated for new AChE 

inhibitors. 

In conclusion, narcissus cultivars are potential sources for AChE inhibitors. As 

these plants commonly contain the known active compound galanthamine, the 

dereplication step of galanthamine is crucial. This step can be done by using the 

on-line HPLC-UV-MS-biochemical detection that has already been developed, or 

by other techniques that could lead to the rapid separation and identification of 

the active compounds from complex mixtures. From our studies, it is clear that 

screening on the level of cultivars using such methodology may result in new compounds 

with AChE inhibitory activity. 


The authors would like to acknowledge the van Leersum Fund for the financial 

support for an HPLC pump.

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16 Narcissus lectins 

Els J.M. Van Damme and Willy J. Peumans 

INTRODUCTION 

Many plants accumulate so-called ‘lectins’, ‘agglutinins’ or ‘haemagglutinins’ in 

their seeds and vegetative tissues. Lectins can be defined as carbohydrate-binding 

proteins which recognise and bind specifically and reversibly to certain mono- or 

oligosaccharides without altering the structure of the bound ligand (Peumans and 

Van Damme, 1995; Van Damme et al., 1998a). In this way lectins are also able to 

interact with glycans of different glycoconjugates such as glycoproteins, glycolipids 

and oligo- or polysaccharides. 

Since the discovery of the first lectin in extracts from castor beans by Stillmark 

(1888), numerous agglutinins have been detected. The rapid progress made in 

biochemical research, and especially the development of very efficient and highly 

specific affinity chromatography techniques for the purification of lectins, has 

resulted in the isolation of a steadily increasing number of agglutinins from varying 

sources. At present over two hundred plant lectins have been isolated and 

characterised in some detail with respect to their molecular structure, biochemical 

properties and carbohydrate-binding specificity. Evidence has accumulated that 

lectins occur in seeds as well as in vegetative tissues of many plant species, and are 

widespread in a large number of plant families belonging to all major taxonomic 

groups (Van Damme et al., 1998b). However, because of the obvious differences in 

molecular structure and sugar specificity between the plant lectins that have been 

characterised thus far, one has to conclude that plant lectins have only a single 

common property, namely their ability to recognise and bind to carbohydrates. 

Until recently, plant lectins have been considered as a very heterogeneous group 

of proteins widely differing from each other with respect to their biochemical 

properties and biological activities. Poor insight into taxonomic relationships 

within the heterogeneous group of plant lectins was mainly due to the lack of 

detailed sequence information. Therefore, the rapid progress in molecular cloning 

and structural analysis of numerous lectins and lectin genes during the last 

fifteen years provided, a powerful means to re-address the question of the classification 

of plant lectins. Based on the available sequence data seven distinct families 

of structurally and evolutionary related proteins are distinguished, which comprise 

the great majority of all currently known lectins. These seven lectin families are 

the legume lectins, the chitin-binding lectins containing hevein domains, the 

monocot mannose-binding lectins, the type 2 ribosome-inactivating proteins, 

the amaranthins, the jacalin-related lectins and the Cucurbitaceae phloem lectins 

(Van Damme et al., 1998a).

Narcissus lectins 381 

This chapter will focus on the lectins present in different species and cultivars of 

Narcissus (daffodil). As will be shown below, narcissus lectin has been studied in 

detail and is shown to be a typical representative of the family of monocot mannosebinding 

lectins. After a brief description of the occurrence of the lectin in different 

Narcissus species and cultivars and the purification of the lectins therefrom, the 

most important findings on the molecular characteristics, carbohydrate-binding 

properties and biological activities of the narcissus lectin will be summarised. 

Furthermore, we will elaborate on the possible function of the lectin in planta as 

well as on possible applications of the lectin in biomedical and glycoconjugate 

research. Finally, a comparison of the narcissus lectin to other known plant lectins 

will be made. 

OCCURRENCE OF LECTINS IN DIFFERENT NARCISSUS 

SPECIES AND CULTIVARS 

The occurrence, isolation and characterisation of a lectin in bulbs of Narcissus was 

first described in 1988 (Van Damme et al., 1988). Since then lectin activity has 

been detected in more than 25 species and varieties of Narcissus (Table 16.1; Van 

Damme and Peumans, 1990a). 

Table 16.1 Narcissus Species and Cultivars with Lectin Activity a 

Botanical species, etc 

Narcissus × medioluteus (N. biflorus) 

N. bulbocodium subsp. bulbocodium var. conspicuus 

N. jonquilla 

N. nanus (N. lobularis) 

N. × maclaeyi 

N. moschatus 

N. nanus 

N. obvallaris 

N. poeticus var. physaloides 

N. pumilus 

N. poeticus var. recurvus 

N. tazetta 

Cultivars derived wholly or partly from: 

N. poeticus: ‘Glory of Lisse’ 

‘Horace’ 

‘Ornatus’ (N. ornatus) 

N. cyclamineus: ‘Bartley’ 

‘Peeping Tom’ 

N. jonquilla: ‘Bobbysoxer’ 

‘Rugulosus’ (N. odorus rugulosus) 

N. pseudonarcissus: ‘Carlton’ 

‘Codlins and Cream’ 

‘Colleen Bawn’ 

‘Empress’ 

‘Queen of Spain’ 

‘W.P. Milner’ 

Other: ‘Double Campernelle’ 

Note 

a Van Damme and Peumans (1990a); names in parenthesis are synonyms.

382 E.J.M. Van Damme and W.J. Peumans 

PURIFICATION OF NARCISSUS LECTINS 

Since preliminary experiments with crude extracts from Narcissus indicated that 

they contained an agglutinating factor that can be inhibited only by mannose, a 

purification scheme based on affinity chromatography on immobilised mannose 

was developed to purify the lectin. Although the affinity purified lectin was virtually 

pure, as could be judged from sodium dodecyl sulphate – polyacryl amide gel 

electrophoresis (SDS-PAGE), additional hydrophobic interactions chromatography 

and ion exchange chromatography were included to ensure complete purity of 

the lectin (Van Damme et al., 1988). The same protocol was used to purify the 

lectin from different species and cultivars of Narcissus. Furthermore, the protocol 

was also applicable for the purification of the lectin from different tissues of 

narcissus, e.g., bulbs, leaves and ovaries. 

CHARACTERISATION OF NARCISSUS LECTINS 

Molecular structure 

The molecular structure of narcissus lectin was determined using SDS-PAGE, gel 

filtration and ultracentrifugation. Upon SDS-PAGE the lectins isolated from all 

Narcissus species and cultivars yielded a single polypeptide band of 12.5 kDa. Since 

the results were identical when electrophoresis was conducted either in the presence 

or absence of β-mercaptoethanol, it can be concluded that the lectin subunits 

are not held together by disulphide bonds (Van Damme et al., 1988). 

Gel filtration experiments on a Superose 12 column using a phosphate buffer 

containing 0.2 M mannose (to avoid binding of the lectin to the matrix) demonstrated 

that the lectins isolated from narcissus elute with an apparent molecular 

mass of 25 kDa, indicating that they are probably dimers. Because aspecific interactions 

of lectins with the gel filtration matrix often occur and cannot be abolished 

completely by the addition of the specific sugar to the running buffer, the molecular 

mass was also determined by ultracentrifugation of the lectins. Since the 

molecular mass of the lectin was calculated to be 36 kDa after centrifugation, the 

lectin may be a trimer (Van Damme et al., 1995). 

It should be indicated here that all lectins isolated from different species and 

cultivars of Narcissus show the same molecular structure and biochemical characteristics 

(E. Van Damme, unpublished data). 

Detailed analyses of the narcissus lectins have shown that the amino acid 

composition of these lectins is typified by high contents of asparagine/aspartic acid, 

threonine, glycine, serine, glutamine/glutamic acid and leucine. The lectins 

contained no amino sugar and only low levels of neutral sugars, most probably 

contaminants, indicating that the lectins are not glycosylated (Van Damme et al., 

1988, 1991a). 

Narcissus lectins are complex mixtures of isolectins 

When the purified lectins from different Narcissus species were analysed by high 

resolution ion exchange chromatography it became evident that they all yielded a


very complex elution pattern, indicating that the lectin preparations are mixtures 

of isoforms (Van Damme et al., 1988; Van Damme and Peumans, 1990a). The 

occurrence of multiple isolectins was confirmed by isoelectric focusing, where the 

purified lectins yielded an extremely complex pattern of polypeptide bands of 

different intensity, indicating that many isolectins were present, some in higher 

concentrations than others. Molecular cloning and sequence analysis of different 

cDNA clones encoding the lectin from Narcissus ‘Fortune’, combined with Southern 

blot analysis, further revealed that the different isoforms were encoded by 

different genes which differed slightly from each other in their sequences (see 

below; Van Damme et al., 1992). 

A detailed study of the isolectin composition in different species and cultivars of 

Narcissus revealed pronounced inter- and intraspecies differences in the isolectin 

patterns. Furthermore, analyses of lectin preparations isolated from different 

tissues at different developmental stages indicated that the isolectin composition is 

both tissue-specific and developmentally regulated. Finally, it was shown that related 

cultivars show similar isolectin patterns (Van Damme and Peumans, 1990a). 

Developmental regulation of lectin concentration 

A detailed study of the developmental changes and tissue distribution of the lectin 

in Narcissus ‘Carlton’ using a very sensitive enzyme-linked immunosorbent assay 

(ELISA) has shown that the lectin occurs in almost all plant tissues, where it is 

present in a very high concentration at the beginning of the growing season (Van 

Damme and Peumans, 1990b). 

In the bulb, the lectin accounts for 10 to 15% of the total tissue protein during 

the dormant phase. However, as the shoot starts to grow the lectin concentration 

in the bulb rapidly decreases. At flowering time almost all lectin has disappeared 

from the bulb. By the end of the growing season the outer bulb scales have been 

degraded. By this time new bulb units have been formed inside the bulb, and by 

the end of the growing season they accumulate high concentrations of lectin as 

they expand. 

High lectin concentrations are also found in the aerial plant parts of narcissus. 

However, the lectin concentrations in the aerial parts are about one order of magnitude 

lower than in the bulb. As the shoot emerges from the bulb, lectin concentrations 

in leaves, stems and flower parts gradually decrease. By flowering time 

almost all lectin has disappeared from these tissues. 

Molecular cloning of Narcissus lectin 

In order to clone the narcissus lectin a cDNA library was constructed from 

poly(A)-rich RNA isolated from young developing ovaries, which are known to 

contain high concentrations of lectin. Screening of the cDNA library constructed 

with mRNA isolated from Narcissus ‘Fortune’, using a previously isolated cDNA 

encoding snowdrop (Galanthus nivalis) lectin (Van Damme et al., 1991b), resulted 

in the isolation of multiple lectin cDNA clones (Van Damme et al., 1992). Although 

the lectin clones showed a high degree of overall sequence homology within their 

coding region, they clearly differed from each other in their nucleotide sequences 

and deduced amino acid sequences. All cDNA sequences contained an open


Figure 16.1 Schematic representation of the biosynthesis, processing and topogenesis 

of the narcissus lectin (NPA). The primary translation product Prepro- 

NPA is processed in the endoplasmic reticulum (ER) by co-translational 

removal of a signal peptide. Upon transport of Pro-NPA to the vacuole a 

C-terminal propeptide is removed, resulting in mature NPA. 

reading frame encoding a preprolectin. This precursor contained, besides the 

coding sequence of the mature lectin of 109 amino acids, a signal peptide (24 

amino acids) and a C-terminal peptide (30 or 38 amino acids) which are co- and 

post-translationally removed, respectively (Figure 16.1; Van Damme and Peumans, 

1988). The calculated molecular mass of the mature narcissus lectin polypeptides 

varied between 11.6 and 11.9 kDa, which is in good agreement with the molecular 

mass of 12.5 kDa determined by SDS-PAGE (Van Damme et al., 1988). 

Interestingly, some differences in the deduced amino acid sequences of the 

different cDNA clones resulted in different charges along the lectin polypeptides, 

resulting in isoelectric points ranging from 3.66 to 4.24 for the mature polypeptides 

encoding the narcissus lectin. These results indicated that the different cDNA 

clones encoded isolectins with different isoelectric points. Hence the detailed 

sequence analysis of different cDNA clones explained the occurrence of multiple 

isolectins at the molecular level. Furthermore, since Southern blot analysis of 

genomic DNA isolated from narcissus yielded numerous restriction fragments 

hybridising with the lectin cDNA probe, it was evident that the expression of 

the isolectins is under the control of a family of closely related lectin genes 

(Van Damme et al., 1992). 

Three-dimensional structure of the Narcissus lectin 

A detailed analysis of the sequence encoding the mature narcissus lectin revealed 

that the sequence shows strong homology with the sequence of snowdrop lectin, 

and likewise is also composed of three very homologous domains (Figure 16.2), 

each of which contains a carbohydrate-binding site (see below). 

Molecular modelling of the deduced amino acid sequence of the mature 

narcissus lectin (derived from the cDNA sequence) using the co-ordinates of the 

related snowdrop lectin as a model, allowed the determination of the threedimensional 

structure of the narcissus lectin (Barre et al., 1996). Like the snowdrop 

lectin, the three-dimensional structure of the narcissus lectin (Figure 16.3) corresponds 

to a β-barrel built up of three antiparallel four-stranded β-sheets (domains) 

interconnected by loops (Hester et al., 1995). A detailed comparison of the 

snowdrop and narcissus lectin sequences revealed that the residues forming the


Figure 16.2 (A) Deduced amino acid sequence of the narcissus lectin precursor encoded 

by cDNA clone LECNPA1. The arrow indicates the cleavage site of the 

signal peptide. (B) Internal sequence similarity of different segments of the 

mature narcissus lectin sequence of 109 amino acids. The amino acids 

composing the carbohydrate-binding site are indicated in bold. 

Figure 16.3 Three-dimensional structure of the narcissus lectin monomer. Strands of 

antiparallel β-sheet are represented by arrows. Asterisks indicate the location 

of the monosaccharide binding site. 

mannose-binding sites are perfectly conserved and that the narcissus lectin 

subunits also contain three functional carbohydrate-binding sites. Hence, as in 

snowdrop, the narcissus lectin monomer possesses three mannose-binding sites, 

each composed of four amino acid residues, Gln, Asp, Asn and Tyr, that bind O2


(Asp and Asn), O3 (Gln) and O4 (Tyr) of mannose through a network of four 

hydrogen bonds. Another hydrophobic residue, namely Val, interacts with C3 and 

C4 of mannose through hydrophobic interactions. 

Recently the narcissus lectin has been crystallised, and its crystal structure in 

complex with α1,3 mannobiose has been determined by X-ray crystallography at 

2 Å resolution (Sauerborn et al., 1999). 

CARBOHYDRATE-BINDING PROPERTIES 

The carbohydrate-binding specificity of the narcissus lectin was assessed by quantitative 

inhibition, sugar hapten inhibition assays using a series of simple sugars, 

and affinity chromatography of glycoconjugates on the immobilised lectin (Kaku 

et al., 1990). Of all monosaccharides tested, only D-mannose was inhibitory in hapten 

inhibition assays. A more detailed study of the carbohydrate-binding specificity 

of narcissus lectin revealed that it had the highest affinity for both terminal and 

internal α1,6-linked mannosyl residues. The narcissus lectin also strongly precipitates 

several yeast mannans (e.g., Saccharomyces cerevisae and Pichea pastoris mannans 

containing multiple D-mannosyl side chains attached to the α1,6-linked 

mannose backbone) but does not precipitate α-D-glucans. Since oligosaccharides 

are better inhibitors than the methyl-α-D-mannoside (e.g., α1,6-linked mannotriose 

being twice as good an inhibitor as Manα1,6Manα-O-Me, and ten times better 

than methyl-α-D-mannoside), it can be concluded that the lectin possesses an 

extended binding site complementary to at least three 1,6-linked α-mannosyl units 

(Kaku et al., 1990). Glycosylasparagine glycopeptides containing α1,6-Man units 

were retarded on a column with the immobilised narcissus lectin. However, glycopeptides 

with hybrid type glycans were not retarded. Therefore the lectin will 

prove to be a useful tool for the detection and preliminary characterisation of 

glycoconjugates. 

It should be indicated that the analyses of the carbohydrate-binding specificity 

of the narcissus lectin were performed with a total lectin preparation containing 

several isolectins. Affinity chromatography experiments suggested differences in 

affinity of different isolectins for a mannose-Sepharose 4B column. Indeed, affinity 

chromatography of the lectin from Narcissus ‘Fortune’ on a column of immobilised 

mannose and elution of the lectin with a linear gradient of increasing mannose 

(0–0.2 M) revealed that some isolectins desorbed from the column at low concentrations 

of mannose, whereas other isolectins were still retained on the column 

after washing with 0.2 M mannose (Van Damme et al., 1990a). These results clearly 

suggest differences in affinity for mannose among the different isoforms of the 

narcissus lectin. 

BIOLOGICAL ACTIVITIES 

The narcissus lectins readily agglutinate rabbit erythrocytes but are completely 

inactive towards human red blood cells, irrespective of the blood group. Agglutination 

of the rabbit red blood cells is enhanced after treatment of the erythrocytes 

with trypsin. The minimum concentrations required for agglutination of


untreated and trypsin-treated rabbit red blood cells were 1.25 and 0.25 µg/ml, 

respectively (Van Damme et al., 1988, 1991a). 

Analysis of the mitogenic activity of the narcissus lectin towards human lymphocytes 

revealed that the lectin is virtually non-mitogenic towards human mononuclear 

cells within the concentration range of 1–200 µg/ml (Kilpatrick et al., 1990). 

In vitro tests demonstrated that the mannose-specific narcissus lectin shows activity 

against human immunodeficiency virus (HIV). The narcissus lectin inhibits 

infection of MT-4 cells by HIV-1, HIV-2 and simian immunodeficiency virus (50% 

effective concentration being 0.5–0.6 µg/ml; Balzarini et al., 1991; Weiler et al., 

1990) at concentrations comparable to those for dextran sulphate inhibition of 

these viruses. Unlike dextran sulphate, the narcissus lectin did not inhibit the replication 

of other enveloped viruses except that of human cytomegalovirus. Furthermore, 

the lectin suppresses syncytium formation between persistently HIV-1 

or HIV-2 infected HUT-78 cells and uninfected MOLT-4 cells (Balzarini et al., 

1991). The narcissus lectin was also shown to prevent rabies virus attachment to 

susceptible cells, and affect rubella virus multiplication after the attachment step 

(Marchetti et al., 1995). 

The lectin was also tested for its toxicity to insects when incorporated into an 

artificial diet, as part of a search for the possible function of the lectin in the plant. 

The narcissus lectin shows antimetabolic effects towards nymphal stages of the rice 

brown planthopper Nilaparvata lugens (Powell et al., 1995) and the peach-potato 

aphid Myzus persicae (Sauvion et al., 1996). Insect feeding trials with the narcissus 

lectin showed that it exhibits a significant antimetabolic effect towards third instar 

nymphs of the rice brown planthopper, although it is less active than the Galanthus 

nivalis lectin, the LC50 for the narcissus lectin being 11 µM compared with 4 µM for 

the snowdrop lectin (Powell et al., 1995). Similarly, addition of the narcissus 

lectin to artificial diets in a concentration range of 10–1500 µg/ml to test the 

toxicity and growth-inhibitory effects on nymphal development of the peachpotato 

aphid revealed that, although narcissus lectin does not induce significant 

mortality, its addition to the diet at 1500 µg/ml resulted in growth inhibition 

(Sauvion et al., 1996). 

PHYSIOLOGICAL ROLE OF NARCISSUS LECTIN IN PLANTA 

The concentration of lectin in various tissues throughout the life cycle of narcissus 

was studied by Van Damme and Peumans (1990b). This study revealed that the 

lectin is present in almost all plant tissues, representing as much as 10% of the total 

protein content during certain stages of development. This is in contrast to most 

of the other plant lectins studied thus far, since their distribution is in most cases 

confined to one or a few tissues and their concentrations are much lower. Since in 

narcissus, as in other Amaryllidaceae species, the changes in lectin content of the 

old and new bulb units coincides with the loss and accumulation of storage compounds, 

it has been suggested that these lectins may function as storage proteins 

which are rapidly degraded as the shoot starts to grow. Furthermore, these lectins 

are present in large quantities in a typical storage organ, the bulb, supporting this 

hypothesis. Most probably these lectins must be considered as storage proteins 

which, as well as their storage role, have an additional carbohydrate-binding


activity. Since the lectin is present only at certain stages of development and occurs 

in almost all plant tissues, it might also play an active role in plant defence. For 

example, the recent discovery that snowdrop and narcissus lectins exhibit toxicity 

towards some insects certainly points in this direction. At present the Amaryllidaceae 

bulb lectins are considered as storage proteins that can also be mobilised as 

defence proteins whenever the plant is attacked by phytophagous invertebrates 

(Peumans and Van Damme, 1995; Van Damme et al., 1998a). The occurrence of 

multiple isoforms may equip the protein with a broad spectrum of biological activity. 

APPLICATIONS OF NARCISSUS LECTINS 

Because of their unique and exclusive carbohydrate-binding specificity, lectins 

have become very useful tools in glycoconjugate research. In this respect, plant 

lectins are often used for the isolation and fractionation of glycoproteins and for 

the study of oligosaccharides and glycopeptides (Osawa and Tsuji, 1987; Cummings, 

1997; Peumans and Van Damme, 1998). Furthermore, lectins are important 

probes in histochemistry and histopathology for the detection of specific carbohydrates 

on cells or in tissue sections (Schumacher et al., 1991). An important prerequisite 

for the successful application and exploitation of a lectin is its commercial 

availability. At present the purified narcissus lectin and preparations derived 

therefrom (immobilised lectin, labelled lectin) are available from at least four companies 

(EY Laboratories Inc., Sigma Chemical Company, Vector Laboratories Ltd. 

and Leuven Bioproducts). 

Hitherto, the narcissus lectin has been successfully applied in the purification of 

glycoproteins and characterisation of glycopeptides on the surface of cells and/or 

proteins. For instance, it was shown that the immobilised narcissus lectin can be 

used for the purification of human α 2 -macroglobulin (Van Leuven et al., 1993). 

As already mentioned above, the narcissus lectins have a strong inhibitory effect 

on the infection of target cells by retroviruses including HIV and cytomegalovirus 

in vitro, which makes them very interesting probes in the study of surface components 

of some viruses. Based on the highly specific interaction between glycoprotein- 

120 (gp120) and the narcissus lectin, an ELISA was developed to determine the 

concentration of gp120 in HIV-infected CEM cells in vitro (Weiler et al., 1991). 

COMPARISON WITH OTHER LECTINS 

The narcissus lectin is a typical representative of the group of monocot mannosebinding 

lectins which was first studied in snowdrop (Galanthus nivalis) (Van Damme 

et al., 1987). Later it was shown that lectins with similar characteristics and biological 

properties also occur in other species of Amaryllidaceae, e.g., Narcissus, Hippeastrum 

and Clivia (Van Damme et al., 1988, 1994). It is now clear that this group of 

lectins is not confined to Amaryllidaceae species, since examples of lectins isolated 

from species belonging to the plant families Alliaceae, Orchidaceae, Araceae, Bromeliaceae 

and Liliaceae clearly show DNA and peptide sequence similarity to the 

Amaryllidaceae lectins. Hence all these lectins are now considered as members of 

the family of monocot mannose-binding lectins (Van Damme et al., 1995, 1998a,b).


All monocot mannose-binding lectins are composed of mature lectin polypeptides 

of 11–14 kDa. Like the narcissus lectin, many other monocot mannose-binding 

lectins are complex mixtures of isolectins resulting from the expression of a family 

of closely related genes (Van Damme et al., 1998a). All DNA sequences encoding 

these lectins show a high degree of homology, resulting in a highly conserved 

three-dimensional structure for the different lectins (Barre et al., 1996; Hester et al., 

1995; Chantalat et al., 1996; Wright et al., 1997). 

Hapten-inhibition assays with simple sugars have demonstrated that all currently 

known monocot mannose-binding lectins are exclusively inhibited by mannose 

(Van Damme et al., 1995). However, the mannose concentrations required for an 

efficient inhibition are high (IC50 =20–200 mM), suggesting that these lectins have 

a low affinity for the monosaccharide. The ability of the monocot mannose-binding 

lectins to distinguish D-mannose from D-glucose units distinguishes this family 

of lectins from the mannose/glucose-binding lectins belonging to the family of legume 

lectins and from the mannose/maltose-binding lectins of the family of jacalinrelated 

lectins (Van Damme et al., 1998a). 

Despite the fact that all monocot mannose-binding lectins will react with mannose, 

they differ in their fine carbohydrate-binding specificity and interaction with 

oligo- and polysaccharides containing D-mannosyl groups. In their immobilised 

form the lectins will also show different chromatographic behaviour towards 

glycosyl-asparagine glycopeptides (Kaku et al., 1990, 1991). 

GENERAL CONCLUSIONS 

At present the lectin is one of the very few proteins from narcissus that has been 

studied in detail. The lectin represents an important fraction of the total protein 

content, at least in resting bulbs. Developmental regulation of the lectin concentration 

and toxicity of the lectin towards insects indicates a possible involvement of 

the lectin in storage metabolism and/or plant defence. 


This work was supported in part by grants from the Katholieke Universiteit 

Leuven and the Fund for Scientific Research Flanders. E.J.M.V.D. is Postdoctoral 

Fellow and W.J.P. a Research Director of this fund. We would like to thank Prof. 

P. Rougé and Dr. A. Barre for their help with the figures. 

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17 Narcissus in perfumery 

HISTORY 

Christian Remy 

The olfactory qualities of the narcissus flower have made it a valuable component 

of luxury perfumes since time immemorial. Originally the flower perfume was 

extracted by the method of enfleurage, in which the substance was absorbed by 

animal fat. The fat was then washed with alcohol, filtered and concentrated to 

obtain a material called absolue des pommades. Later, extraction was carried out by 

using a solvent, generally hexane, which allowed better extraction and faster 

processing. This procedure, equally applicable to other flowers such as rose, 

jasmin, orange-blossom, etc., was perfected towards the middle of the nineteenth 

century, but it was at the start of the twentieth century that it began to be used on 

an industrial scale. 

When first used, wild narcissus flowers were harvested in Provence, in the 

hinterland of the Côte d’Azur, where the growing of lavender, lavandin and sage 

is now predominant. Narcissus grow in grassy areas (Figure 17.1), and the gradual 

decline in livestock farming in the region and its replacement by arable farming 

resulted in the scarcity of the flower, to a point at which its harvesting was no 

longer profitable. From 1950, it has been necessary to look elsewhere for supplies. 

The manufacturers in Grasse, who already knew the southern Auvergne well for 

its lichen (tree moss) growing on forest pine, also turned towards this area for 

narcissus (Narcissus poeticus) and Jonquil (N. jonquilla). Jonquils are used in perfumery 

as well as narcissus, but in smaller amounts. The Auvergne is traditionally 

a land of flower, herb and mushroom gathering, indeed the very phrase ‘flower 

gathering’ evokes a complimentary, almost trivial activity, or in any case an activity 

on the fringes of agriculture. But it is nothing of the sort here, if one takes into 

account the volumes involved and their commercial importance. In the Auvergne, 

flower gathering must be considered as a vital source of revenue, and a real part of 

the overall rural economy of the region. But there was a time when flower gathering 

was done just for the sake of it, as much as for sale, selling at the fair what was 

surplus to the family’s needs. 

A major change came about at the end of the Second World War, when there 

was a rise in flower gathering specifically for sale. In many cases, and especially 

with medicinal plants, this was linked with the development of the pharmaceutical 

industry, but other crops have remained stable right to today. It was at this time 

that plant collection was organised into a network, leading to the advent of 

collectors and gathering teams. As well as narcissus, they also gather violets, wild

Narcissus in perfumery 393 

Figure 17.1 Narcissus (Narcissus poeticus) can be seen like white sheets in pastures. On 

the right of the photograph there are some Jonquils (N. jonquilla), which 

generally flower 2–3 weeks before the narcissus. Both flowers can be 

found in the same area, but they never blend. 

anemone, arnica, St. John’s wort, wild pansy, mallow, burdock, cat’s foot, foxglove, 

balm and gentian. Today, all these products have become part of a more modest 

flower gathering operation, sometimes very small scale, and among some of the 

principal products its scale is strictly controlled: bilberry and mushroom on the 

one hand, and tree moss, narcissus and jonquils on the other. The last three products 

are virtually the only materials of the region destined for use in perfumery. 

The narcissus, the jewel in the crown of the region’s plants, has become its 

emblem. In the last days of May, the meadows suddenly become white in successive 

waves, reaching ground as far as 1500 metres above sea level. Narcissus generally 

grow in damp meadows, and the intensity of the white covering that forms in the 

fields can reliably indicate the presence of water or a stream from a distance. 

FLOWER GATHERING 

Thirty years ago, some flower picking was still done by hand, but even at that time 

the use of the ‘comb’ (le peigne) was becoming more common (Figure 17.2). The comb 

was a kind of rake with cutting teeth, and it was designed to allow the collection of 

just the flower head on its own, without the stalk. In fact, it is only the centre of the 

flower itself that is of interest in perfumery. Gradually, one saw appearance of a 

larger, improved comb, a cart mounted on wheels (le chariot). The cart is somewhat 

cumbersome, but is more efficient than the comb on large areas lush with flowers; 

however, it is not particularly suited to all situations, and the comb continues to be

394 C. Remy 

Figure 17.2 Flower collecting implements: the comb (right) and the cart (left). 

used for small isolated patches of flowers or on sloping terrain where agility is 

needed. The principle of the cutting teeth is the same on both tools. The cart 

allows the cutting of three-four times as many flowers as the comb. 

As stated above, narcissus grow in meadows, which can apparently lead to conflicts 

of interest. Unlike most forests where tree moss is collected, and which are 

public places, the meadows are privately owned. Meadows are always carefully 

fenced, since they are used for livestock farming, and this systematic fencing, 

added to the temperament of the Auvergne farmers, reinforces the fact of ownership. 

Two situations are possible: 

• Where the owner has authorised access to his field for picking (possibly sharing 

the profit with the picker). 

• Where access has been prohibited since the owner plans to crop it himself, 

because he believes that, in view of his decision not to exploit the field, there is 

no reason for somebody else to earn money from it. 

This reasoning perhaps appears to have an odd logic, but it is still frequently 

encountered. If it is only a small part of a field that is covered with flowers, it is 

very tempting to leave it to waste! These conflicts can sometimes lead to violence, 

and, each year, some settling of scores are ended by lead shot. Sometimes there is 

even clandestine picking, at night. Thus, a landowner who one evening sees before 

his very eyes a field of white flowers, can find a green field the next morning! 

As one can imagine, one of the problems posed by the gathering of narcissus is 

the fragility of the flower. More so than in the case of tree moss, which is not 

perishable, narcissus gathering demands a network of strict organisation because


collection is carried out in an area 200 km across and 500 km from Grasse. It is 

therefore practically impossible for a flower destined to be treated at Grasse to 

arrive in a satisfactory state of freshness. Picking, collection and transport by truck 

takes two to three days, and the flowers are largely degraded during this length of 

time. For this reason, a central extraction plant has been set up right in the heart 

of the collection area, allowing the treatment of flowers on the same day that they 

are collected. Thanks to this proximity, the quality of a product has been transformed 

and greatly improved. The central facility alone processes 50–60% of the 

total flowers collected each year. 

QUANTITIES 

The gathering of narcissus flowers is, as is the case for most plant material, subject 

to the weather. This situation is perhaps accentuated by the fact that flowering 

takes place in mountainous regions, at altitudes between 600 and 1500 metres. 

Thus, cold weather persisting in the preceding months, or late frosts, can jeopardise 

flowering. Collection may equally be spoilt in years with strong rain, because 

of the excessive, rapid growth of grass, practically covering the narcissus flowers 

and making cropping impossible in some places. However, it is true that, even if 

the flowers are poor, in theory there will be more than enough to satisfy the needs 

of the perfume industry. The problem is simply that the pickers, after having evaluated 

the density of flowering and having carried out a rapid calculation of profitability 

by the hour, decide to crop . . . or not to crop. Their decision will depend on 

whether collection levels that year will be normal or low. The quantities gathered 

will also be adjusted according to the needs indicated by the industry. This is why, 

with medium sized crop being between 300 and 400 tonnes, quantities can vary 

between 200 and 600 tonnes in different years. These are the weights of flowers 

alone, excluding the stems and leaves. 

EXTRACTION 

Narcissus flowers are fragile and delicate. It is desirable to treat them as soon as 

possible after collection, in order to preserve their olfactory qualities. While waiting 

for treatment, the flowers are spread out on a cement slab at the factory (Figure 

17.3). They are stored for as little time as possible, in a thin layer which is turned 

over constantly to avoid fermentation starting. As is the case for the flowers of rose, 

jasmin, orange blossom and many others, they are treated by extraction with solvents 

(hexane being the usual solvent), in order to obtain a viscous product called a 

‘concrete’ (Figure 17.4). The concrete contains a high proportion of vegetable waxes 

that render it useless, in that state, in perfumes. It is, therefore, necessary to remove 

these waxes in order to render the product soluble in alcohol. The concrete is a 

thick substance that hardens on cooling, and it has to be heated in a bain-marie to 

soften before being processed. The concrete is subjected to a second transformation 

through washing with alcohol in order to separate the waxes and to obtain 

the final product, referred to as an ‘absolute’. It is the absolute that is used in 

the manufacture of perfumes. The narcissus belongs to the category of noble

Figure 17.3 Narcissus flowers spread out to dry, and a drum holding 20 kg of concrete, 

the equivalent of 10 tonnes of flowers. 

FLOWERS 

Extraction with solvent (hexane) 

(0.2% yield) 

CONCRETE 

Washing with alcohol 

(37% yield) 

Removal of insoluble 

vegetable waxes 

ABSOLUTE 

Figure 17.4 Scheme for the production of narcissus absolute.


perfumes, a title shared by other flowers such as rose, jasmin and bitter orange 

blossom. 

The low productivity of absolute from the narcissus makes it an expensive product 

that cannot be used other than in luxury perfumes: 

• 1000 kg of flowers provides about 2 kg of concrete (0.2%). 

• From this amount of concrete, after removing the waxes, 750 g of absolute is 

obtained. 

It therefore takes a total of 1300–1400 kg of flowers to obtain 1 kg of absolute. 

However, it is not only its price that determines the value of narcissus absolute, but 

also the olfactory quality, and these factors determine the success of a perfume. 

OLFACTORY DESCRIPTION 

It is always difficult to describe an olfactory sensation in words. Our vocabulary is 

just not adapted to it, whatever language we use. It is also very difficult to know if 

one particular scent is perceived in the same manner by two different people. All 

this makes the description of a scent a difficult and hazardous exercise. In the case 

of narcissus, an olfactory description is so much more difficult since this absolute is 

almost a perfume by itself. Its scent is rich and complex. In it one can sometimes 

be reminded of the elusive fragrance of iris, rose, jasmin, tuberose, ylang-ylang, 

orange blossom, storax and oak moss. All these brought together form a scent that 

is intense and lasting, flowery and exciting at the same time. 

The main components of the volatile part of narcissus absolute are listed in 

Table 17.1. 

Table 17.1 Main Components of the Volatile 

Part of Narcissus Absolute 

Compound % of Volatile 

α-terpineol 23.7 

Methyl trans-isoeugenol 20.0 

Benzyl benzoate 19.4 

Coumarin 8.9 

Benzyl alcohol 5.0 

δ-3-carene 3.4 

Phenylethyl alcohol 2.2 

Ethyl palmitate 2.2 

Cinnamyl alcohol 2.0 

Phenylpropyl acetate 1.7 

1,8-cineole 1.5 

β-caryophyllene 1.0 

Benzyl acetate 0.7 

Isoeugenol 0.6 

cis-3-hexenyl acetate 0.5 

Cinnamyl cinnamate 0.5 

Total 91.3

398 C. Remy 

CONCLUSION 

The use of narcissus in perfumery has to take account of practical realities. Modern 

perfume making is very cost-conscious, and much emphasis is placed on the 

appearance of the packaging. Perfumes have to be sold in the face of ever increasing 

competition, and the advertising budget necessary for launching a perfume 

swallows up considerable sums. Then, far too often, manufacturers economise on 

the quality of the perfume. The imagination of the perfumier is also constrained, 

and much of the time he is forced to exclude the more expensive components from 

his formula. In perfumery, these are often the choicest ingredients. 

This is why one has to say that the use of narcissus in perfumery is rather in 

decline. One can hope, in the years ahead, to see a reversal of this trend, when it 

will be realised afresh that the success of a perfume depends on its quality. The 

secret of the really great perfumes is that they endure all fashion trends. 

All the information in this chapter has been based on experience gained at 

Laboratoire Monique Remy. Some additional sources of information may be 

found in the “Further reading” section. 

FURTHER READING 

Bruno, S., Laurentis, N. de, Amico, A. and Stefanizzi, L. (1994) Chemical investigation and 

cytologic localisation of essential oils in the flowers of Narcissus tazetta. International Journal 

of Pharmacognosy, 32, 357–361. 

Dobson, H.E.M., Arroyo, J., Bergstrom, G. and Groth, I. (1997) Interspecific variation in 

floral fragrances within the genus Narcissus (Amaryllidaceae). Biochemical Systematics and 

Ecology, 25, 685–706. 

Dort, H.M. van, Jagers, P.P., Heide, R. ter and Weerdt, A.J.A. van der (1993) Narcissus 

Trevithian and narcissus Geranium: analysis and synthesis of compounds. Journal of Agricultural 

and Food Chemistry, 41, 2063–2075. 

Ehret, C., Maupetit, P. and Petrzilka, M. (1990) New organoleptically important components 

from Narcissus absolute (Narcissus poeticus L.). In Proceedings 11th International Congress of 

Essential Oils, Fragrances and Flavors, New Delhi, November 1989, S.C. Bhattacharyya, 

N. Sen and K.L. Sethi (eds.), Vol. 5, Aspect Publishing, London, pp. 49–55. 

Ehret, C., Maupetit, P. and Petrzilka, M. (1992) New organoleptically important constituents 

of narcissus absolute (Narcissus poeticus L.). Journal of Essential Oil Research, 4, 41–47. 

Lawrence, B.M. (1995) Progress in essential oils. Perfumer and Flavorist, 20, 35–48. 

Lawrence, B.M. (1997) Progress in essential oils. Perfumer and Flavorist, 22, 59–67. 

Li, B.L., Lu, B.Y., Luo, Y.J., Chen, Z.H. and Zhu, L.F. (1992) GC/MS analysis of the 

fragrances of five narcissus. Acta Botanica Austro Sinica, 8, 165–172 (in Chinese). 

Loo, A. and Richard, H. (1988) A study in Narcissus absolute composition. In: Proceedings 

10th International Congress of Essential Oils, Fragrances and Flavors, Washington, November 

1986, B.M. Lawrence, B.D. Mookherjee and B.J. Willis (eds.), Developments in Food Science, 

18, 355–373.

18 Harmful effects due to Narcissus 

and its constituents 

Celia G. Julian and Peter W. Bowers 

INTRODUCTION 

The genus Narcissus has been grown commercially in the far south-west of England 

for more than a hundred years. The warm climate and small sheltered fields 

(particularly on the Isles of Scilly) have helped this industry, for which significant 

expansion appears likely in the next few years. Many hundreds of seasonal workers 

are employed, the larger labour force being needed for 6–8 weeks in the spring 

when the flowers are picked, bunched and packed for distribution. A smaller 

number of seasonal workers are employed in summer for lifting and cleaning the 

bulbs for re-planting or sale. Thus, any adverse effects from contact with narcissus 

(daffodil) can influence a potentially large number of workers and have a significant 

sequel. 

At present, more than 1180 ha are planted with narcissus in south-west England, 

which includes a significant expansion within the last two years. This provides a 

considerable revenue for Cornwall and the Isles of Scilly, areas which otherwise 

rely on farming, fishing and tourism and which have a high rate of unemployment. 

Cornwall accounts for about 35% of the bulb acreage in the UK, other significant 

areas being in Lincolnshire, Norfolk and Scotland. 

EFFECTS ASSOCIATED WITH HANDLING NARCISSUS CROPS 

Handling flowers 

‘Daffodil itch’ or ‘lily rash’ associated with picking narcissus is well recognised and 

was first described by Walsh in 1910 in relation to flower pickers on the Isles of 

Scilly. The name ‘lily rash’ arose as narcissus were originally thought of as lilies, 

although they are now placed in the Amaryllidaceae because they have an inferior 

ovary. An eczematous rash develops from direct contact between the skin of the 

picker and sap from the flower stem. Granulomatous sores may develop at any 

point of direct trauma and paronychia can occur. These changes may be quite 

disabling, but as the season is short and the rash therefore self-limiting, the pickers 

rarely present to a doctor. Hands and wrists are the most commonly affected 

areas, but the rash may also appear beneath the chin, on the forearms, in the axillae 

and on the genitalia. 

The way in which the flowers are picked and collected accounts for the distribution 

of the rash. Picking takes place when the flowers are still in bud. The picker

400 C.G. Julian and P.W. Bowers 

Figure 18.1 Daffodil picking: snapping off the flower stem. 

Figure 18.2 Granulomatous rash on the wrist. (See Colour plate 3) 

slides a hand down to the base of the flower stem and then snaps, cuts or pulls off 

the stalk (Figure 18.1). During this process, the finger webs are particularly vulnerable 

to trauma by the ends of the daffodil leaves or by stalks from previously 

picked flowers. The base of the stalk continues to grow after the flowers have been 

cropped, resulting in an increasing hazard to the pickers as each row may be 

sequentially picked up to four times, with more protruding stalks on each occasion. 

As the flowers are gathered, sap drips out from the stems onto the wrist and 

forearm, causing further rash (Figure 18.2).

Figure 18.3 Daffodil picking: gathering up the bunches. 

Harmful effects of Narcissus 401 

On the mainland daffodils are cropped mainly in the January to March period. 

The flowers are picked by one hand and transferred to the other until ten have 

been collected. The ends of the stems are then levelled against the palm of the 

hand and the bunch is secured with an elastic band from a supply kept in a pot at 

the waist. The picker straddles the row of plants and works along it, laying the 

bunches down in groups of three or five for collection on return. Working back 

down the row, the bunches are gathered in the crooks of the arms and under the 

chin and armpits (Figure 18.3), to be deposited in a personally labelled tray. This 

is a rapid procedure, as an experienced operator is capable of collecting in excess 

of 2000 bunches per day (20 000 flowers). The money earned is related directly to 

the number of bunches picked. Workers are told at the beginning of the day the 

price payable per bunch. They therefore have a financial incentive to hold on to as 

many bunches as possible. This practice is reflected in the more extensive distribution 

of the rash. 

On the Isles of Scilly, the earlier flowering Tazetta varieties are grown, cropping 

from October onwards. The method of flower picking is the same as on the 

mainland, but the flowers are carried by tractor to sheds for bunching and 

packing. The pickers are paid an hourly rate, rather than by the number of 

bunches gathered, and do not need to hold large quantities of flowers in close 

bodily contact. On the Islands, the primary daffodil rash is therefore usually 

confined to the hands and wrists. In both groups of workers, secondary rash may 

develop on the face (Figure 18.4), or on the scrotum in men as a result of transfer 

of daffodil sap by direct contact with the hand while urinating.


Figure 18.4 Secondary facial rash. (See Colour plate 4) 

Apart from ‘daffodil itch’, there are other medical problems associated with 

picking narcissus flowers. The workers are perpetually bent forwards, with their 

backs to the wind, and are prone to backache. All pickers run the risk of having 

repetitive strain injury to the wrist, from the twisting action as the flower stem is 

plucked from the plant. The more inexperienced pickers sometimes wear inappropriate 

clothing, and can develop chilblains on the backs of their thighs and 

buttocks (Julian and Bowers, 1997). 

Handling bulbs 

Bulb lifting takes place during a six week period in June/July. Between 12 and 

20 tonnes of bulbs are planted per hectare. After two or three years, depending on 

the variety, the yield of bulbs should be at least twice the planted weight, bulb 

quality is at its peak, and the bulbs are harvested. With a total UK crop area of 

over 4000 ha, huge numbers of bulbs are involved. In Lincolnshire, where the 

land was reclaimed from the sea and there are no large stones in the soil, the 

lifting process is often largely mechanical. Lifting machines (complete harvesters) 

lift the bulbs and transfer them to a hopper, removing much of the soil in the 

process, although workers on the lifting machine may remove clods. The bulbs are 

then transported to sheds for grading and further treatment. In Cornwall, where 

the soil contains large stones and the fields are often small and steeply sloping, the 

lifting process is only partly mechanical, the bulbs being elevated by a lifting 

machine to the surface where they are left to dry for several days before being 

gathered into baskets by hand and transferred to large boxes. Seasonal workers 

are again employed to pick up the bulbs, making the procedure more labourintensive 

in Cornwall than in Lincolnshire. In Cornwall the bulbs are dry when 

picked and are handled relatively little, and in practice this gives few problems to 

those that collect them.

Prevention of daffodil rash 


Protective clothing is an important aspect in the prevention of daffodil rash. 

Experienced workers are well aware that they should minimise any contact 

between daffodils and the skin. Flower picking often takes place during wet and 

windy weather. Waterproof clothing is essential to prevent sap soaking through to 

the skin as the bunches are collected. The majority of pickers invest in good 

rubber, or even nitrile, gloves, and are aware that they must avoid any gap 

between the end of the glove and the sleeve of their jacket, and elastic bands or 

neoprene cuffs are utilised for this. Bulb lifting takes place in the summer months, 

generally under drier conditions. The workers universally wear gloves, but need 

encouragement to provide adequate protection from the sun by wearing a hat and 

applying sunscreen regularly. 

The rash resulting from direct contact between daffodil sap and the skin of susceptible 

individuals is mainly a primary irritant dermatitis. Experienced workers, 

picking up to 20 000 flowers per day, have potential contact with a large volume of 

sap. The amount of sap produced varies between cultivars, and also within the 

same cultivar under different conditions, as direct observations confirmed (Julian 

and Bowers, 1997). More sap is produced during wet weather. Pickers commented 

to the present authors that the ability of a daffodil to produce an irritant rash varies 

with the variety. The highly scented, multi-flowered ‘Soleil d’Or’ and ‘Paperwhite’ 

Tazetta narcissi, grown almost exclusively on the Isles of Scilly, rarely cause any 

problem. The trumpet varieties ‘King Alfred’ and ‘Princeps’ were mentioned 

specifically by older flower farmers on the Isles of Scilly. These cultivars are not 

grown in quantity at the present time, but were notorious in the past for the problems 

they caused. Pickers are aware of these differences: when gathering Tazetta 

narcissus they hold the flower stems facing inwards, but when picking standard 

varieties they turn round the stems, to avoid sap dripping onto their clothing. 

Constituents of Narcissus sap 

Narcissus sap contains calcium oxalate crystals, which are polymorphic and may be 

multi-faceted cubes or needle-shaped (Figure 18.5). The needle-shaped crystals 

are grouped in bundles known as raphides. A study by Sakai et al. (1984), which 

did not include narcissus, demonstrated an increase in irritancy in species with 

long raphides. With the electron microscope, it was possible to determine the 

presence of barbs and grooves in the crystal structure. Their presence was associated 

with irritancy. In their absence, only crystals with a length exceeding 180 µm 

produced irritation in Sakai’s study. The relationship between crystal appearance 

and sap viscosity to irritancy in a number of narcissus cultivars is the subject of a 

current study by the present authors. 

In narcissus the calcium oxalate crystals are contained within ‘slime vessels’. 

These are formed by the elongation of normal parenchymatous cells and the subsequent 

breakdown of their dividing cell walls. As well as calcium oxalate crystals, 

they contain mucilage and alkaloids. When the flower stalk is twisted off during 

picking, the slime vessel is ruptured and its contents extruded by the force of 

turgidity of the adjacent intact cells (J.J. Beijer, personal communication). In this 

way, calcium oxalate crystals are able to penetrate intact skin (van der Werff, 1959).


Figure 18.5 Polymorphic calcium oxalate crystals. The needle-shaped crystals are 

ca. 100 µm in length. (See Colour plate 5) 

The alkaloids found in the slime vessels may be responsible for the rarely occurring 

allergic reactions to narcissus occasionally reported (Hausen and Oestmann, 

1988). Traumatisation of the skin by calcium oxalate crystals allows the penetration 

of these substances from the slime vessels. At least 15 different alkaloids have 

been isolated from narcissus (Barton et al., 1963), but only two of these were found 

to be capable of producing a weakly positive allergic response when injected into 

sensitised guinea pigs (Gude et al., 1988). These alkaloids, homolycorin and masonin, 

were not detected in the stems or leaves of narcissus: their highest concentration 

was found in the bulbs, although sap extracted from bulbs produced minimal 

allergic effect. It would seem from this work that narcissus has very little ability to 

produce an allergic response. Additionally, a Finnish study produced only two


patients with allergic contact dermatitis to narcissus over a fourteen-year period 

(Lamminpää et al., 1966). Both were gardeners with many years’ exposure, and 

produced positive patch tests to a 1% concentration of narcissus plant extract. 

During discussions with workers in the Cornish narcissus industry, the present 

authors met a farm owner who developed an immediate facial erythema and swelling 

on entering the packing sheds, as a result of sensitisation from previous handling 

of the flowers. 

Pesticides 

Pesticides are important in the flower industry and worldwide have been noted to 

produce increasing skin problems in agricultural workers (Lisi et al., 1987). This 

is predominantly a contact dermatitis where protective measures are poor. It has 

been reported in the tulip bulb industry, after use of the fungicide fluazinam. In 

this case the bulbs were handled after spraying, contrary to the manufacturer’s 

instructions (Bruynzeel et al., 1995). This fungicide is not used in the daffodil bulb 

industry in the UK, where the list of fungicides includes chlorothalonil, carbendazim, 

iprodione, vinclozolin, benomyl and mancozeb, and no particular problems 

have been observed with these. On one farm where we have observed bulb processing, 

carbendazim is currently used, but all bulb dipping takes place in tanks 

outside, not in bulb sheds. The same material is also used as a spray in the field to 

control fungal foliar diseases and delay leaf senescence. Fungicides may be present 

on the leaves at the time of picking, but field workers questioned by us did not 

report any particular problem with this. However, some farmers reported they 

were unable to spray the crops because of the severity of the facial rash which they 

develop while doing so. This is allergic dermatitis to airborne particles which contain 

the fungicide to which they have become sensitised. No specific product was 

mentioned. 

Conclusions 

Our study of those involved in the narcissus industry has shown that most harmful 

effects are produced as a result of direct contact between irritant sap and the skin. 

Many pickers are affected, but they have developed ingenious strategies to avoid 

this contact and allow them to work. The season is short and the condition selflimiting, 

so they rarely present to a doctor and the problem is under-reported. 

OTHER HARMFUL EFFECTS OF NARCISSUS 

Ingestion by humans 

A rare but harmful effect of narcissus is toxicity as a result of the inadvertent 

ingestion of the bulbs (Venner and Gibbons, 1995). When mistaken for small 

onions and included in a stew, acute vomiting resulted from the effects of toxic 

alkaloids, including lycorine which is stable to heat. Recovery was rapid, due to 

the early onset of severe emesis. This probably accounts for human toxicity being 

limited to vomiting, abdominal cramps, shivering and diarrhoea. The onset of


symptoms is rapid, but usually resolves spontaneously within 3 hours (Litovitz 

and Fahay, 1982). In a second report (Vigneau et al., 1984), nausea persisted in 

one subject for 10 days. However, all involved were completely recovered 15 days 

later. 

Effects on other cut-flowers 

The presence of daffodils in a vase with other cut-flowers has a deleterious 

effect on them (Barendse, 1974; van Doorn, 1998). This effect was very noticeable 

with tulips, whose flower stems droop and wilt within a few days, as was 

shown experimentally by Gugenhan (1970) who studied the effects of placing 

narcissus and tulip in separate vases and together. Narcissus and tulip kept 

separately remained normal, but within 6 days tulips placed with narcissus had 

deteriorated. While the narcissus still looked healthy, the tulips had aged, with 

mottling of the leaves and curvature and wilting of the flower stems. A similar 

but lesser effect was seen if tulips were placed in water previously occupied by 

narcissus, suggesting that the daffodil sap, rather than the actual flowers, was 

harmful. This effect was noted by Sytsema and Barendse (1975) to vary between 

cultivars, the sap of ‘Carlton’ being particularly aggressive. From a study with 

different flower foods, Terfrüchte (1981) reported that one sort could prevent 

this damage to tulip flowers. It would, however, seem advisable to confine all 

cut-flowers of narcissus to their own vase and refrain from mixing them with 

other species. 

Blankenship and Richardson (1987) attempted to identify the component of 

narcissus sap responsible for causing damage in tulips. Over a 2-year period they 

studied the effects of ethylene, ethylene inhibitors and auxins, and concluded that 

the major senescence effect of stem curvature was caused by auxins. Auxins have 

been isolated both from the plant and from the water in which narcissus have been 

placed, and may be released into the water to produce their deleterious effect on 

flowers of other species. In their earlier study of narcissus auxins, Edelbluth and 

Kaldewey (1976) had found that auxin-inhibitory activity occurred in diffusates of 

stem segments, buds and flowers, and could be removed by washing to reveal 

auxin activity. Since narciclasine is present in narcissus in considerable quantities 

and is strongly inhibitory to cereal seedling growth (Ceriotti, 1967; Bi et al., 1998), 

this could be the material interfering in the bioassays used to detect auxins. Van 

Doorn (1998) later investigated the deleterious effects of narcissus mucilage on 

cut-flowers of tulip and rose. Fractionating the narcissus mucilage, this author 

suggested that the effect on roses was due to the sugar and polysaccharide fraction, 

which resulted in increased bacterial growth and blocked water uptake, while 

the effect on tulips was due to the toxicity of a fraction that contained several 

alkaloids. 


The authors would like to acknowledge the help of Allen Scrimshaw and family, 

Roger Rosier and Ian Wort in the preparation of this work.

REFERENCES 


Barendse, L.V.J. (1974) Schade door narcisseslijm bij verschillendse bloemsorten. Vakblad 

voor de Bloemisterij, 29 (2), 12–13. 

Barton, D.H.R., Kirby, G.W., Taylor, J.B. and Thomas, G.M. (1963) The biogenesis of 

Amaryllidaceae alkaloids. Journal of the Chemical Society, Part 6, 4545–4558. 

Bi, Y.R., Yung, K.H. and Wong, Y.S. (1998) Physiological effects of narciclasine from the 

mucilage of Narcissus tazetta L. bulbs. Plant Science, 135, 103–108. 

Blankenship, S.M. and Richardson, D.G. (1987) Auxin–like activity of daffodil exudate 

reduces vase life of cut tulips. Gartenbauwissenschaft, 52 (3), 130–135. 

Bruynzeel, D.P., Tafelkruijer, J. and Wilks, M.F. (1995) Contact dermatitis due to a new 

fungicide used in the tulip bulb industry. Contact Dermatitis, 33, 8–11. 

Ceriotti, G. (1967) Narciclasine: an antimitotic substance from Narcissus bulbs. Nature, 213, 

595–596. 

Doorn, W.G. van (1998) Effects of daffodil flowers on the water relations and vase life of 

roses and tulips. Journal of the American Society for Horticultural Science, 123, 146–149. 

Edelbluth, E. and Kaldewey, H. (1976) Auxins in scapes, flower buds, flowers, and fruits of 

daffodil (Narcissus pseudonarcissus L.). Planta, 131, 285–291. 

Gude, M., Hausen, B.M., Heitsch, H. and König, W.A. (1988) An investigation of the irritant 

and allergic properties of daffodils (Narcissus pseudonarcissus L., Amaryllidaceae). 

Contact Dermatitis, 19, 1–10. 

Gugenhan, E. (1970) Schnittblumen – Haltbarkeitsversuch. Der Erwerbsgärtner, 24, 656–657. 

Hausen, B.M. and Oestmann, G. (1988) Untersuchungen über die Häufigkeit berufsbedingter 

allergischer Hauter frankungen auf einem Blumengrossmarkt. Dermatosen in Beruf 

und Umwelt, 36, 117–124. 

Julian, C.G. and Bowers, P.W. (1997) The nature and distribution of daffodil pickers’ rash. 


Lamminpää, A., Estlander, T., Jolanki, R. and Kanerva, L. (1996) Occupational allergic 

contact dermatitis caused by decorative plants. Contact Dermatitis, 34, 330–335. 

Lisi, P., Caraffini, S. and Assalve, D. (1987) Irritation and sensitisation potential of pesticides. 


Litovitz, T.L. and Fahay, B.A. (1982) Please don’t eat the daffodils. New England Journal of 

Medicine, 306 (9), 547. 

Sakai, W.S., Shiroma, S.S. and Nagas, M.A. (1984) A study of raphide microstructure in 

relation to irritation. Scanning Electron Microscopy, 2, 979–986. 

Sytsema, W. and Barendse, L. (1975) Houdbaarheid snijbloemen krijgt steeds meer aandacht. 

4. Vakblad voor de Bloemisterij, 30 (49), 16. 

Terfrüchte, J. (1981) Narzissen nun doch in ‘Bunten Strausse’. Flora Bric gegen Narzissenschleim. 

Gärtnerbörse unt Gartenwelt, 81, 305–306. 

Venner, E. and Gibbons, D. (1995) The Lady of Shallot. The Remedy – Cornwall Clinical 

Journal, 4 (1), 11. 

Vigneau, C., Tsao, J., Ducluzeau, R. and Balzot, J. (1984) Two clinical cases of daffodil 

poisoning. Journal of Toxicology and Medicine, 4 (1), 21–26. 

Walsh, D. (1910) Investigation of a dermatitis amongst flower pickers in the Scilly Islands, 

the so-called ‘Lily Rash’. British Medical Journal, 2, 854–856. 

Werff, P.J. van der (1959) Occupational disease among workers in the bulb industries. Acta 

Allergologica, 14, 338–355.

19 Review of pharmaceutical patents 

from the genus Narcissus 

James R. Murray 

INTRODUCTION 

This chapter is to give the reader a flavour of the richness of concepts associated 

with the Narcissus genus whilst bowing to the practicalities of reporting complicated 

patent processes that evolve over a period of years. No attempt is made to 

give definitive lists of published versus granted patents, as such lists would, by 

necessity, be out of date by the time of publication of this book. Instead, the 

chapter uses the listings of published patents and applications as of mid-September 

1999 as a basis of a comprehensive review of the areas of research and development 

of products and projects associated with the cultivation and utilisation of 

narcissus (daffodil) plants and their constituents. 

On undertaking the search for patents published around the genus Narcissus, 

28 were identified under the title ‘narcissus’, 11 under ‘daffodil’ and 55 under 

‘galanthamine’. As galanthamine was originally extracted from the Caucasian 

snowdrop it was, perhaps, not surprising that only two of the 39 patents identified 

under ‘narcissus’ or ‘daffodil’ referred to galanthamine. Although not strictly 

related to Narcissus species, these patents are included here for completeness. The 

‘narcissus’ and ‘daffodil’ titles are mainly related to use in cosmetic agents and 

fragrances, and many of these patents are from Japan and without equivalents 

outside Japan. These patents refer to synthetic molecules designed to mimic narcissus 

fragrance, as well as extracts of the plant. The galanthamine patents cover 

manufacture, intermediates used in the production of galanthamine, analogues of 

galanthamine, and the use of the cholinesterase inhibitory and nicotinic agonist 

pharmacological activities of the compounds in the clinic. Such clinical indications 

range from the original Eastern European folklore use of galanthamine in the 

management of patients with infantile paralysis (poliomyelitis) to the treatment of 

erectile dysfunction! 

NARCISSUS AND DAFFODIL PATENTS 

Patents related to non-therapeutic uses 

A fascinating array of patents describing the utilisation of agents derived from 

daffodils, to mimic the fragrance of daffodils, or for agents or methods used to 

influence daffodils themselves, was uncovered. Who could fail to be intrigued by 

the grappling shovel with twin concave blades mounted on handles, operated with

Narcissus patents 409 

the aid of a foot pedal that has been developed for optimisation of bulb, corm or 

tuber planting? Or by the fact that it is possible, by fusion bonding a laminate of 

two sheets of hot-melt type adhesives with dry pressed flowers (including daffodils 

as an example) in between, to form a decorative sheet-laminated fabric? 

Oscillating digging shears for bulb lifting, with the front undercutting shear 

oscillating in the opposite phase to the following lift section, and processes to turn 

waste flower bulbs into chipboard by a pressing treatment at high temperatures 

and pressures (utilising the starch, cellulose and water in the bulbs), featured with 

an intriguing family of patents relating to optical and electronic uses, including 

endoscope diagnostic systems, in the patent search relating to the title ‘narcissus’. 

The latter optical patents referred, in fact, to ‘the narcissus type radiation effect’ – 

clearly not related to the genus but more to the original Narcissus himself! These 

patents bloated the original ‘narcissus’ hit list by 15! 

Other patents relating to the inhibition of stem elongation in flowering bulbous 

plants using 1-amino-cyclopropane-1–1carboxylic acid synthase inhibitors, or to 

methods of protecting foliage from browsing deer by spraying with deer-repellant 

extracts of narcissus bulbs, contrasted somewhat with a series of patents for hair 

and skin cosmetics containing waxes (Krause et al., 1998), ‘sedative essential oil 

comprising of narcissus absolute oil’ (Kanebo Ltd., 1994) or a ‘new’ ortho-methyl 

cinnamic acid phenylethyl ester perfume (Hofmannor et al., 1989) said to have a 

‘flowery aroma’. Such ‘flowery aromas’, if containing terpenic fragments, can further 

be protected from degradation, according to another patent, by adding rosemary 

and/or sage components! Other fragrances include 6-acyloxy-hexanoate 

esters (Ochsner, 1986) and 4-hydroxy or alkanoyl 3-ethoxy benzyl alkyl ethers, 

which have a ‘narcissus aroma’ (Ochsner, 1985). The plant-derived waxes 

are hydrophobic and biodegradable, and can be used as hair lotions, soaps or 

shampoos. The waxes are extracted from the flowers and form a protective film on 

the skin or hair. They are also said to improve ‘combability’ of hair. Daffodil 

fragrances are variously described as ‘warm’, ‘fruity’ or ‘floral’ and are often used to 

augment ‘woody’ or ‘musk’ aspects of perfumes. 

Lectins derived from a number of plants, including the genus Narcissus, have an 

anti-nematode activity, causing mortality, reduced larval weight and/or delayed 

development, whilst being non-toxic to animals and birds. They can be used to 

protect a variety of crops including grain, cotton, potatoes, sugar cane, tomatoes, 

etc. (Birch et al., 1995). 

It was not surprising to find patents protecting the use of a medium containing 

abscisic acid in facilitating the multiplication of bulbs by tissue culture, or the 

method of encouraging vegetative bulb multiplication by cutting the bulb prior to 

deposition into substrate. The problems of policing some of these patents may be 

fraught with difficulties! 

Therapeutic uses 

The therapeutic uses of agents derived from daffodils under the narcissus/daffodil 

titles also included those that could be regarded as ‘cosmeceuticals’. However, of 

particular interest are patents referring to the potential anti-viral activity of a 

mannose-specific lectin from, for example, Narcissus pseudonarcissus. A vaccine, produced 

from antibodies raised in vivo or in vitro to the lectin, is particularly relevant

410 J.R. Murray 

to certain RNA viruses, which contain glycoproteins with mannose linkages, 

including Human Immunodeficiency Virus (HIV) and Human T lympho-tropic 

Virus (HTLV), either as a therapy or as a diagnostic (Forrest et al., 1991). 

In addition, the use of bulb extract with vinegar has been claimed for the treatment 

of fungal diseases of the skin, in particular, tinea pedis (Sugimoto, 1994). 

Topical treatments derived from hot-water extracts of ground daffodil bulbs are 

claimed to dilate peripheral veins, improve blood flow to improve metabolism and 

thus complexion, and also to prevent skin ageing and damage to skin by ultraviolet 

light (Kobayashi, 1982). 

PATENTS RELATED TO GALANTHAMINE 

Galanthamine patents fall into four groups – production, formulation and use 

patents and galanthamine analogues. The parent molecule has been well categorised 

many years ago, so compound patents on the molecule itself do not feature in 

the literature. However, compound patents around analogues and derivatives of 

galanthamine and precursors of galanthine do. 

Production patents 

Introduction 

Galanthamine was first extracted from members of the snowdrop family (hence 

galanthamine from the genus Galanthus). Commercially, daffodil bulbs are far 

more appropriate, as they are available in bulk supplies. Extraction from daffodils 

in the UK has been particularly relevant, as some 70% of the world’s supply of daffodil 

bulbs come from the British Isles, with growers in the east of England playing 

a major role. Handling large volumes of bulbs has its problems. McFarlan Smith 

(Meconic Ltd.) in Edinburgh now has one of the largest plant extraction units in 

Europe dedicated to galanthamine extraction. 

Clearly with bulb production being rather a slow process, the extraction method 

does lack some flexibility, and much effort has been put into exploring methods of 

synthetic production. Unfortunately, the galanthamine molecule has three chiral 

centres with eight possible enantiomers. In practice, one of these centres is effectively 

‘locked’ and thus four stereoisomers are possible. However, the plant produces 

only one stereoisomer, the (–)-galanthamine, and it is this isomer that is most 

active pharmacologically. Synthetic production problems lie in producing the one 

active isomer and in overcoming a crucial oxidative phenyl-coupling step that has 

a particularly low yield. 

Extraction patents 

Given that extraction technologies contain as much art as science, it is not surprising 

that only a few patents exist around this method of production. However, such 

patents do exist, particularly from Eastern European authors, with the starting 

material sometimes involving sources other than daffodils. For example, the roots


and bulbs of the ‘sunflower Galanthus Kzasnovii’ (sic) are said to give an extraction 

galanthamine yield of 0.5% (dry weight) (Asoyeva, 1968). 

Galanthamine can also be extracted from the summer snowflake Leucojum aestivum 

(Proskurnina, 1961), using an 8% ammonium aqueous solution plus dichloroethane 

or other organic solvents followed by a sulphuric acid treatment, with 

subsequent crystalline precipitation of other alkaloids such as lycorine and thebaine. 

Further treatment of the liquor with chloroform allows for extraction of the 

galanthamine itself by vacuum distillation. 

The snowdrop (Proskyrni, 1960) is the starting material in another patent, the 

final liquor being treated with a solution of hydrobromic acid and acetone to form 

galanthamine hydrobromide. Ground, fresh amaryllis bulbs are moistened with 

7% ammonium hydroxide and extracted with dichloroethane in another patent 

process file that leads to a yellow oily liquid which crystallises on standing 

(Asoyeva, 1963). Another variant of the Leucojum aestivum source involves treatments 

with aluminium oxide in the final stages of extraction (Ordzhonikidze 

Chem-Pharm, 1962). 

A Japanese patent file describes the production of galanthamine, utilising clay 

or ion-exchange resins, from Lycoris squamigera (Shionogi & Co. Ltd., 1960). Extraction 

from the leaves and flowers of Galanthus nivalis (the common snowdrop) is the 

subject of another patent, but the practicalities of isolating from plants that are not 

available commercially in sufficient biomass limit the value of such approaches 

(Chimiko Pharmazetitschen Zavod, 1968). Production from Ungernia victoris 

(Aleksandrova et al., 1994) in tissue culture is also cited as a commercially viable 

process. The claim that by first bringing the extract of bulb derived material to 

pH4 can lead to a pure form of the alkaloid galanthamine, is made in yet another 

publication (Hille et al., 1996). 

Production of synthetic galanthamine 

The chiral nature of the galanthamine molecule with its three asymmetric carbon 

atoms has led to considerable problems in synthetic manufacture. Even though 

processes have been described several decades ago, commercialisation has, until 

recently, been severely limited by low yields. Obtaining an optimal starting point 

for (–)-galanthamine has been the subject of much research. In particular, narwedine 

has been the subject of several patent filings as a potential means of addressing 

the problem. For example, the preparation of a single (–)-narwedine enantiomer 

by seeding a solution of racemic narwedine with single enantiomer (–)-narwedine 

in the absence of amine base, plus the reduction of the resultant (–)-narwedine to 

the single (–)-galanthamine enantiomer is claimed, with the advantage that such 

(–)-narwedine prepared by this method is configurationally stable to racemisation 

in the solid state and can be produced on a large scale with very high purity (Potter 

and Tiffen, 1998). 

Another approach has sought to limit the problems of possible contamination 

with unwanted epi-galanthamine compounds by incorporating specific reducing 

agents in the process of converting (–)-narwedine to (–)-galanthamine (Carlsson 

and Shieh, 1995). 

Attempts to increase the yields in the phenolic coupling stage of the preparation 

of new and known narwedine derivatives include a method in which oxidation


occurs in a two-phase liquid system comprising aqueous base and organic solvent 

of dielectric constant of less than 4.8. The process improves the yield of sufficiently 

pure products in the organic phase to allow recovery by evaporation, avoiding 

relatively expensive chromatographic purification. These products can then readily 

be converted to corresponding galanthamine structures (Henshilwood and Johnson, 

1996). Other filings from the same group describe asymmetric transformation of 

racemic narwedine-type compounds by reacting these racemates with an enantiomerically 

enriched acid to form a diastereomeric salt (Chaplin et al., 1997b). 

A method for ‘easily’ preparing narwedine, lycoramine, norgalanthamine and 

sanguinine by the preparation of intramolecularly coupled compound comprising 

of a phenol derivative with a supervalent iodine reagent is described elsewhere 

(Dyer et al., 1996). As far as galanthamine itself is concerned, similar techniques 

can be used to prepare specific enantiomers, for example by seeding a supersaturated 

solution of racemic galanthamine salt with an enantiomerically enriched 

form of the salt, with or without the utilisation of an achiral counter-ion (Kagaku 

Gijutsu Shinko Jigyodan, 1999). 

Of particular interest, as it has already led to a commercially available source of 

galanthamine, is an Austrian patent which embraces methods of producing new or 

known galanthamine products from benzaldehyde and phenethylamine derivatives. 

By reacting these together and reducing the product, an N-benzylphenylamine 

derivative is formed. This is then subjected to oxidative cyclisation and the 

product is then reduced (Tiffen, 1997). 

A fascinating method of ‘feeding’ galanthamine precursors (derivatives) to plant 

extract to allow the enzymatic conversion by the plant material of the oxidative 

cyclisation precursor, is described. The object is to enhance yields of optically pure 

forms over and above those expected from straightforward extraction techniques 

(Czollner et al., 1995). 

Formulation patents 

These patents fall into two categories – those related to combination therapy and 

those describing galenic presentation forms. 

The combination of an acetyl-cholinesterase inhibitor with a muscarinic agonist 

(citing galanthamine as one of the preferred cholinesterase inhibitors with a variety 

of possible muscarinic agonists quoted) has been claimed to be useful in treating 

central and peripheral nervous system diseases (Bannister and McCague, 1997). 

A fast-dissolving galanthamine hydrobromide tablet is the subject of published 

patent applications. The tablet includes a spray-dried mixture of lactose monohydrate 

and microcrystalline cellulose (75:25) as a diluent with a new disintegrant 

(Callahan and Schwarz, 1999). 

Transdermal delivery of galanthamine is claimed to give better control of 

release of drug over a longer period of time (at least 24 hours), steadier serum levels 

and higher therapeutic effects at lower dosages with better patient acceptability 

(De Conde and Gilis, 1997). The system utilises a reservoir layer of active agent in 

a polymer matrix plus a penetration enhancer. This homogenous reservoir 

mixture is coated onto a backing layer and then covered with a protective layer. 

Interestingly, the same company has filings around the recovery of the active 

agent from unused or discarded transdermal therapeutic systems using solvent


extraction (Deurer and Hille, 1994). Another transdermal delivery system containing 

pergolide free base or its mesylate or hydrochloride salt is also described (Asmussen 

et al., 1997). 

An oral sustained release composition, in the form of a tablet or capsule, incorporates 

the use of active ingredient particles coated with an enteric protection agent, 

polyvinyl pyrrolidone (Fischer et al., 1998). 

New galanthamine analogues or derivatives 

Benzazepine analogues and diaza-bi cycloalkanes (Davis and Goodman, 1994), 

methylapogalanthamine hydrochloride (Czollner et al., 1997) (the latter having 

hypotensive properties), 1-aminomethyl-1-phenyl-4-hydroxy-cyclohexane derivatives 

said to be analgesic in activity (Abdusamatov, 1963), spiro-benzazepines (also 

analgesic and sedative) (Grelan Pharmaceutical KK, 1975) and 6-O-demethylgalanthamine 

(Grelan Pharmaceutical Co., 1975) are amongst a whole host of 

analogues and other salts (Davis et al., 1995a,b,c; Chaplin et al., 1997a; Christen 

et al., 1997) that are claimed to be more potent, or less toxic or more brain-specific 

cholinesterase inhibitors, than galanthamine itself. Some of these patents cover not 

only the novelty of the molecules, but also method(s) of production. 

Specific production patents include means of ‘producing galanthamine derivatives 

by condensing and reducing a benzaldehyde derivative and a tyramine derivative, 

oxidatively cyclising the protected product and reducing the keto group’ 

(Czollner et al., 1996) and another by a ring closure method (Grelan Pharmaceutical 

Co., 1972). 

Use patents 

Patents discussed under this heading are often broader in scope than just being 

related to galanthamine. Analogues, and even cholinesterase inhibitors in general, 

are often also included in these claims. 

The original uses of crude concoctions of snowdrop bulbs and, later on, of more 

highly purified materials, included an array of neuro-muscular disorders, such as 

poliomyelitis and myaesthaenia gravis, as well as more vague neurological conditions. 

The discovery of the reduction of levels of choline acetyl transferase in the 

brains of Alzheimer’s Disease victims led to the search for compounds that could 

cross the blood-brain barrier and boost cholinergic activity in the basal nuclei, hippocampus 

and cortex. Tacrine was the first acetyl cholinesterase, to be developed 

for such a therapeutic use in Alzheimer’s Disease, but it has a greater affinity for 

butyryl than for acetyl cholinesterase, and may lead to raised liver enzyme activity 

that can seriously restrict its use. Galanthamine is one of a number of so-called 

second generation inhibitors that do not have adverse activity on the liver, readily 

pass the blood-brain barrier, and are more specific inhibitors of the acetyl form of 

cholinesterase. In addition, galanthamine has nicotinic activities, which could well 

differentiate it from other second-generation cholinesterase inhibitors in the 

clinic. 

The major indication for galanthamine at present is Alzheimer’s Disease. Oral 

(10–2000 mg/day), parenteral (0.1 to 4 mg/kg) and intracerebroventricular administration 

via an implanted reservoir (0.01–5.0 mg/kg/day) are claimed to improve


cognitive function (Davis, 1987). A later patent was taken out by the same author 

for the same indication and routes of administration, but covering new derivatives 

of galanthamine or their analogues (Davis and Joullie, 1988). 

Patients with Down’s Syndrome (trisomy of chromosome 21) can develop 

histological changes resembling those seen in the brains of Alzheimer’s patients. 

Learning ability in patients with Down’s Syndrome may be improved by galanthamine 

(Fransits and Mucke, 1997). 

One prolific investigator from Iceland has taken out a series of patents addressing 

the treatment of mania (Snorrason, 1994), fatigue syndromes (Snorrason, 

1992a), various forms of arthritis (Snorrason and Murray, 1997), attention deficit 

disorders (Murray and Snorrason, 1999a) and proteolytic diseases (Murray and 

Snorrason, 1999b), as well as methods for countering the side-effects of benzodiazepine 

therapy (Snorrason, 1992b,c). Acute mania is a difficult condition to treat 

and often requires high doses of neuroleptics given for a number of days. The 

author claims galanthamine to be a faster treatment than other drugs. Stress is 

placed within the patent on the activity of galanthamine at nicotinic receptors. 

Fatigue is a common enough symptom in primary care medicine, but its clinical 

importance cannot be over-estimated. It can be a major aspect of significant morbidity. 

Fatigue associated with viral infections, especially the Epstein Barr virus, 

can be prolonged and debilitating. Mononucleosis in late-teens or the University 

years can result in long periods of marked fatigue leading to considerable impairment 

of performance. In its severe form – myalgic encephalitis (ME) or Chronic 

Fatigue Syndrome (CFS) – patients experience debilitating fatigue lasting for 

more than 50% of the day over a period of six months or more. Such patients are 

usually in the ‘bread-winning’ years of life and may be unable to work for several 

years and can even be dependent upon a wheelchair. No effective treatment presently 

exists for this condition. Galanthamine is claimed as a treatment for fatigue, 

muscle pains and sleep disturbances in CFS. Claims are made for galanthamine, or 

its derivatives, to be effective in the treatment of various fatigue syndromes, 

including those associated with HIV infections and pre-eclampsia. 

Cholinesterases also have proteolytic enzyme activity. Two patents from the 

Icelandic author, working with a British colleague, address the potential for galanthamine 

to have a positive effect on disease conditions in which proteolytic 

enzymes appear to play an important role. The first addresses the role of raised 

proteolytic enzyme and cholinesterase activity in the synovial fluid in the joints of 

patients with rheumatoid arthritis. Claims are made for galanthamine in the treatment 

of rheumatoid and osteo-arthritis, arthritis associated with auto-immune 

disease, psoriasis, chronic inflammatory bowel disease, ankylosing spondylitis, as 

well as a range of other joint disorders. Such treatment, if successful, could be 

expected to be disease-modifying rather than simply palliative. The second patent 

addresses head-on the concept that proteolytic activity in other diseases is associated 

with acetyl cholinesterase activity, and that this could be blocked or inhibited 

by treatment with galanthamine, epigalanthamine or norgalanthamine. Such diseases 

would include psoriasis, Crohn’s Disease and ulcerative colitis. 

Attention deficits and hyperactivity disorders (ADHD) are thought to affect 5% 

of children in the USA and Europe. Moreover, such behavioural problems can be 

carried into adult life where they may manifest themselves as alcohol or drug 

abuse and other anti-social behavioural traits. Current therapy in childhood


includes the use of methylphenidate and amphetamines. Claims for the modification 

of these behavioral problems if proved in the clinic could lead to an effective 

therapy that avoids the use of potentially habit forming drugs. (Another patent 

from different authors claims ADHD, but also the rarer and more bizarre 

Tourette’s syndrome, as an indication for galanthamine (Davis, 1999).) 

Counteraction of sedation, hypnosis or respiratory depression seen during 

benzodiazepine therapy, especially when high doses are given, can be achieved by 

co-administering galanthamine or its analogues or derivatives without interfering 

with the anxiolytic, anti-psychotic, anti-convulsant or muscle relaxant activities of 

the benzodiazepines. Particular emphasis is made in one of two patents from the 

same authors stressing the advantage of such combined therapy in patients with 

schizophrenia, but the patent also includes those suffering from anxiety, anxiety 

neurosis, panic disorders and agitating depression, as well as other psychiatric 

conditions. 

Patients suffering from schizophrenia may experience improvement in states of 

apathy and aboulia when treated with 2 mg of amizil 1 to 3 times daily in conjunction 

with injections 30 minutes later of 2 ml of 0.5% galanthamine solution once or 

twice a day according to another patent (Kamenetski and Losev, 1985). 

Alcohol and nicotine dependency can both be favourably influenced by galanthamine 

therapy according to two patents (Oplitz, 1991; Moormann and Moormann, 

1994), who particularly stress the possibility of reducing the desire for 

alcohol or nicotine in addicted patients. These patents particularly, but not only, 

mention the transdermal delivery of galanthamine in such cases. 

Given the early use of galanthamine it is not surprising, but very pleasing, to 

find a reference to treating spastic forms of cerebral paralysis in children with 

galanthamine and other drugs prior to bio-training sessions. Motor function was 

improved by 46% when galanthamine was used, compared with 33% for biotraining 

without galanthamine or 14% when galanthamine was used alone (Bogdanov 

et al., 1984). 

Electrodiagnostic methods for determining the degree of damage to a nerve can 

be improved in terms of accuracy by administering to the patient 1 ml of 1% galanthamine 

in solution a half to one hour prior to testing (Korletyanu, 1986). 

Administering a cholinesterase inhibitor, such as galanthamine, either alone or 

in combination with levodopa, reduces rigidity, improves muscle function and 

alleviates dementia in Parkinson’s Disease (Hutchinson, 1998). 

Acute di:phenyl-hydramine poisoning can be treated more effectively, quicker 

and in a shorter period of time with riboxin (inosine) given together with galanthamine 

(Afanasev and Lukin, 1993). 

A bio-stimulant composed of a lyophilised hydrosylate of royal jelly, creatinine, 

a calcium-magnesium salt of inositol phosphoric acid and galanthamine hydrochloride 

is claimed to be useful for patients recovering from severe or chronic illnesses. 

The combination is also claimed to help persons such as sportsmen or 

soldiers under great physical stress by acting on ‘the energy balance of the body, 

improving the activity of the central and peripheral nervous systems and of 

cardiac and striated muscle’ (Kirov et al., 1983). 

Another patent covers the alleviation of jet-lag, whereby galanthamine is used 

as a means of resetting a person’s internal clock. The compound is given as an 

attention increasing agent to travellers prior to departure. The drug should be


given at a time between 4 a.m. and 3 p.m. of the time zone to be visited. To ‘reset 

the clock’, galanthamine should be administered at least 8 hours prior to the 

desired time of sleep. The authors believe such therapy could also be relevant to 

shift workers (Davis, 1996). 

CONCLUSIONS 

The powers of the genus Narcissus can be seen to be many and varied. Each plant 

should have its specific medical use, and clearly cognitive disorders and Alzheimer’s 

Disease in particular must be the candidates for the daffodil. However, 

various forms of sexual dysfunction that can lead to severe strain on otherwise 

meaningful partnerships, and cause considerable distress to the sufferer, are not 

uncommon and can also be addressed by the noble daffodil. Physiological erectile 

impotence has been the subject of much recent research and has led to effective, if 

presently expensive, therapies. However, the use of galanthamine for erectile 

dysfunction due to physiological causes secondary to other disease states, disorders 

of the urogenital or endocrine systems, due to drug therapy (e.g., anti-hypertensives, 

anti-depressant and anti-psychotic drugs) or related to psychiatric problems, has 

been advocated (Katz, 1993). In addition, the Japanese patent that advocates the 

use of crude extracts of plants, such as Lamium album, Sesamum indicum and 

Narcissus tazetta, together with a number of fatty acids and surfactants to improve 

humectant properties, elasticity and smoothness of ageing skin would add to the 

surmise that daffodils could provide means of addressing the failings of advancing 

years (Taisho Pharm Co. Ltd., 1993). In any event, strong pharmacological 

reasoning, if backed by effective clinical studies, could well mean that the Narcissus 

genus will play an important role in medical and cosmetic practice well into the 

new millennium. 

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Index 

For further alkaloids, readers should also consult Tables 6.1, 6.3 and 6.4; for further species 

and cultivars, Table 6.2; and for further biological and pharmacological effects, Table 6.4. 

Abscesses, 25, 26 

Acetyl-10-norpluviine, 258 

Acetylcholine, 358, 371 

Acetylcholinesterase, 336, 338, 343, 416 

Acetylcholinesterase inhibitors, 195, 215, 

243, 256, 273, 282, 286, 293, 328, 335f, 

339, 371ff, 410, 414f 

Acetylgalanthamine, 258 

Advisory information (bulb growing), 59f 

Agavaceae, 276 

Agricultural subsidies, 135 

Agrochemical use, reduced, 101, 280 

Air pollutants, 97 

Aizoaceae, 141 

Ajax (genus), 30 

Alcohol abuse, 256, 274, 417 

Aldrin, 85 

Alkaloids, 2, 24, 141ff, 215ff, 258, 264, 

273ff, 286ff, 371ff, 405ff; analysis, 249, 

257; biosynthesis, 142, 163, 169, 171, 

179; composition (cultivars), 373; 

distribution, 277; structure, 141f, 185; 

synthesis, 196, 306ff 

Allelopathic, 185 

Allergenic, 185 

Allergic dermatitis, 407 

Allergic reactions, 406f 

Alliaceae, 400 

Allium cepa (see onions) 

Alzheimer’s disease, 25, 195, 215, 243, 

250, 273ff, 279ff, 286, 304, 334ff, 357ff, 

366, 372f, 415, 418 

Amaryllidaceae (in general), 1, 7, 31, 141, 

215, 242, 256ff, 276, 286ff, 371ff 

Amaryllis (Hippeastrum), 244, 290, 

413 

Ambeline, 288, 290 

Amino acid sequences, 386 

Amino-cyclopropane-carboxylic acid 

synthase, 411 

Amyloid plaques, 343 

Anaesthesia, 273, 343, 347 

Analeptic, 195 

Analgesic, 26, 195, 256, 345, 415 

Angel’s Tears daffodils, 38 

Anhydrogalanthamine, 269 

Anodic oxidation, 320 

Antiarrhythmic, 243 

Anticancer, 197, 274 

Anticholinesterase activity, 215, 242ff, 

274, 304, 334ff, 341, 346, 358f 

Antifertility, 185 

Antifungal, 185 

Anti-inflammatory, 196 

Antileukaemic, 351 

Antimalarial, 196 

Antimetabolic, 389 

Antimicrobial, 185 

Antimitotic, 185, 196 

Antinociceptive, 185 

Anti-social behavioural traits, 416 

Antispasmodic, 26 

Antitumor, 185, 196, 350 

Antiviral, 185, 196, 274, 389, 411 

Anxiety, 417 

Apex size, 11 

Aphelenchoides subtenuis, 60, 100 

Aphids, 61f, 81, 88, 389 

Aphrodisiac, 26 

Apical dominance, 10 

Apical meristem, 10, 14 

Araceae, 390 

Arthritis, 416 

Ascorbic acid metabolism, 196 

Asphodel, 21, 24 

Atropine, 342 

Attention deficit and hyperactivity 

disorder, 414 

Auto-immune diseases, 416 

Autumn-flowering species, 4, 31 

Auxins, 4, 217, 408 

Azospirillum spp., 71

420 Index 

Backache, 403 

Bacterial diseases, 61 

Baldness, 26 

Barley, 134 

Basal (base) rot, 59, 61, 68–102 

Basal plate (stem plate), 4, 14 

Basal plate disease, 100 

Beans, 134 

Bees, 13 

Behavioural problems, 416 

Benomyl, 76 

Benzazepines, 243 

Benzimidazole fungicides, 94 

Benzodiazepine therapy, 273, 414f 

Benzylaminopurine, 220ff 

Bible, 23, 25 

Bioavailability, 338 

Biochemical detection system, 374f 

Biological activities, 186ff, 388 

Biological control, 101 

Biology, 1ff 

Biomass, 247, 277ff 

Bio-stimulant, 417 

Black slime, 61 

Blood-brain barrier, 339, 343 

Bombus spp., 13 

Boron, 71 

Botrytis narcissicola (see smoulder) 

Bradycardia, 347f 

Brain injuries, 195, 344 

Breeding, 1, 27, 31, 102ff 

Bromeliaceae, 390 

Broncholytic, 243 

Bruising (of bulbs), 71 

Brunsvigia boophane, 178 

Bud abortion, 2 

Bufadienolides, 215 

Bulb; branching system, 4ff; certification, 

90, 98; cleaning, 93, 97f; clusters, 5, 11; 

dips, 77, 94; drying, 78, 94ff, 100f; forcing, 

55; grade, grading, 11, 64, 82, 93ff; growing 

equipment, 131, 135, 137, 139; growth/ 

development, 5, 280; handling, 93f; 

inspection and sorting, 63, 93, 97f; lifters, 

lifting, 91ff, 281, 411; orientation, 84; 

packing, 97; prices, 55, 132f; quality, 62ff; 

ripening, 90f; scales, 4ff; shapes, 7, 64; size, 

64, 276, 279; source, 62ff, 215; splitting, 97; 

storage, 2, 72f, 77ff, 93, 96ff, 139; structure, 

4ff, 275ff; yields, 91, 132, 279, 281 

Bulb and leaf nematode, 60 

Bulb mites, 60 

Bulb planting, 79, 84f, 411; arrangement, 

84f; band, 84; date, 82; density, 11, 

80ff, 276, 279f; depth, 83ff, 276, 279; 

machines, 79, 84; material, 62ff; nets, 86; 

row orientation, 84 

Bulb producing areas, 55f 

Bulb production, 53ff, 131ff; for processing, 

59, 81, 88ff, 98, 101, 109; seeds, 109; 

small-bulbed types, 100; specialist types, 

99ff; species, 100; sustainable, 1; tazetta 

narcissus, 99f 

Bulb scale mite, 60, 72 

Bulb units, 4ff 

Bulbing, 8, 14 

Bulbocodium group, 30, 47 

Bumble bees, 13 

Butyrylcholinesterase, 336 

Calcium, 71 

Calcium oxalate, 12f, 405f 

Callus culture, 215ff 

Cancer, 24ff, 286, 338 

Captan, 76 

Caranine, 166f, 277 

Carbendazim, 76 

Carbofuran, 78 

Carbohydrates, 2 

Catarrh, 26 

Cats, 26 

Cattle, 26f 

Cauliflower, 134 

Cell cultures, 248 

Cell growth inhibition, 196 

Cell wall structure, 14 

Centrifugation, 287 

Cereals, 85, 135 

Cerebral paralysis, 415 

Chemiluminescence methods, 373 

Chemotaxonomy, 141 

Chemotherapy, 334 

Chilblains, 404 

Chinese Sacred Lily, 42 

Chipboard, 411 

Chipping (propagation), 11, 14, 64, 97, 

103ff, 108, 279, 281 

Chlidanthine, 170 

Chlorpyrifos, 76, 78 

Chocolate spot, 62 

Cholinergic activity, 195, 215, 336, 342, 

371, 414 

Cholinergic hypothesis, 358, 371 

Cholinesterase, erythrocyte, 339 

Chromatography, 249ff; affinity, 382, 384; 

capillary gas, 249, 288, 299, 372; counter 

current, 295; electrokinetic capillary, 294; 

gas, 257, 288ff; HPLC, 216, 249, 257ff, 

289ff, 337ff, 374; paper chromatography, 

249, 291f; reversed-phase HPLC, 290f, 

295; TLC, 290ff 

Chromatography-mass spectrophometry, 

288ff 

Chromosome numbers, 102

Chronic fatigue syndrome, 25, 346, 416 

Chronotropic, 185 

Classified Lists, 50, 62 

Climate, 64ff 

Clivia, 163, 390 

CNS toxicity, 349 

Cognitive function and memory, 337, 361, 

416, 418 

Colchicine, 215 

Colchicum, 215, 226 

Cold hardiness, 65 

Cold period (cold requirement), 2, 4, 14, 60 

Collar daffodils, 47, 49 

Colour code, 47, 50 

Commercial floriculture, 1ff, 11, 53, 80, 

242ff, 246ff, 276 

Common Agricultural Policy, 135 

Common names of daffodils, 21 

Complexion, 412 

Composted waste, 71 

Concrete, 397 

Conspectus, 34ff 

Consumer tastes, 59 

Consumption (eating), 26f, 184, 347, 407 

Contraceptive, 26 

Cooperanthes, 244 

Corbularia, 30 

Corona (paracorolla, trumpet or cup), 1, 13 

Cortisol levels, 348 

Cosmeceuticals, 411 

Cosmetics, 410f, 418 

Costs; bulb growing, 136ff; capital, 137ff; 

fixed, 135ff 

Cover crops, 85f 

Criglaucine, 290 

Crinamidine, 295 

Crinine, 142, 168f, 290, 295 

Crinine/haemanthamine-type alkaloids, 

168ff, 178, 181 

Crinum, 244, 288ff, 294f, 

Criwelline, 182 

Crohn’s Disease, 414 

Crop; covers, 65, 85f; duration, 11, 53, 73, 

79ff, 105ff, 132f; inspection, 90; rotation, 

68f, 280 

Crop handling; bulbs, 404, 406; flowers, 

401, 407 

Crops, common, 134 

Cross-cutting (propagation), 106 

Crusades, 24 

Cultivars, 31, 47, 49, 57, 59, 62, 102, 275f, 

280, 381; choice, 57; collections, 62; 

double, 13, 49; dwarf, 76f, 82, 100; groups, 

57, 289; most grown, 57ff; trials, 62 

Cultivars (names); Carlton, 11, 59, 61, 85, 

102, 257, 271, 289, 375, 385, 408; 

Cheerfulness, 59, 279f; Deanna Durbin, 

Index 421 

166; Dutch Master, 59; Fortune, 10, 59, 65, 

248f, 257, 388; Geranium, 279f; Gigantic 

Star, 257; Golden Harvest, 11, 59, 61, 85, 

102f; Hawera, 88; Ice Follies, 59, 249, 257, 

279ff; Inglescombe, 276ff, 282; King Alfred, 

10, 163, 166f, 169, 405; Mount Hood, 279; 

Odorus Rugulosus, 245; Princeps, 405; 

Sempre Avanti, 166; Sir Winston Churchill, 

376; Soleil d’Or, 42, 99, 105, 405; St. 

Keverne, 102; Tête-à-Tête, 58, 59, 65, 77, 

85ff, 100, 106; Texas, 163; Ziva, 99 

Curare, 342 

Cut-flowers, phytotoxic effects, 405, 408 

Cyclamineus narcissus, 31, 57, 74, 100 

Cytogenetics, 102 

Cytokinins, 4, 197, 218ff 

Cytology, 31 

Cytotoxic, 185 

Daffodil itch or rash, 401, 404f 

Daffodil, derivation of name, 21f 

Damage to bulbs/crops; flower cropping, 

87; formaldehyde, 61, 75; herbicides, 61, 

87; hot-water treatment, 60, 75ff; 

mechanical, 61, 97 

Datura stramonium, 334 

Daughter bulbs, 5 

Daylength, 14 

Deer-repellant, 411 

Defoliation, 91f 

De-heading (flower removal), 88f, 279 

Dehydrogenase activity, 196 

Demethylgalanthamine, 287, 291, 341, 350 

Demethylgalanthamine carbamates, 282 

Demethylgalanthamine glucuronide, 341 

Demethyl-lycoramine, 277, 282 

Demethylmasonine, 258 

Demethylnarwedine, 170, 282 

Deoxynarciclasine, 295 

Deoxy-trans-dihydronarciclasine, 294 

Depression, 417 

Dermatitis, 26, 407 

Desiccants, 92 

Detectors; electron impact, 288, 290; flame 

ionization, 299; nitrogen phosphorus, 

299; ultra-violet, 289, 291, 293f, 299 

Dextromoramide, 345 

Diacetyl-10-norpluviine, 258 

Dichlorophenoxyacetic acid, 216 

Dichloropropene, 70 

Dihydrolycorine, 243, 274 

Dimethylnorbelladine, 171, 308 

Disease diagnostics, 69 

Diseases (general), 60ff, 83 

Disinfectants, 72, 75 

Disorders (plant and flower), 60ff 

Ditylenchus dipsaci (see stem nematode)

422 Index 

Divisions (classification), 47ff 

Donepezil, 359 

Dormancy, 2, 4, 59f, 276ff 

Down’s syndrome, 416 

Drugs (general), 280, 341, 345, 416 

Dysentry, 26 

Dysmenorrhoea, 345 

Eastern Europe, 273, 335, 341ff, 410 

Ecology, 2 

Economics of bulb production, 69, 131ff 

Electrodiagnostic methods, 417 

Electro-encephalography, 337, 339 

Electrophoresis, 294 

Electrophoresis (SDS-PAGE), 384 

ELISA testing, 64 

Embrocation, 26 

Emetic, 26, 185 

Enantiomers, 412ff 

Endangered species, 1, 27 

Endomycorrhizal fungi, 71 

Enfleurage, 394 

Environmental implications, 100 

Enzyme immunoassay, 276, 293, 337, 372, 

385, 390 

Epigalanthamine, 269, 291, 305, 339ff, 

413, 415 

Epihaemanthamine, 168 

Epilepsy, 26 

Epilycorine, 180 

Erectile dysfunction, 410, 418 

Erysipelas, 26 

Essential oils, 185 

Ethylene, 4, 12, 15, 97, 226, 408 

Eumerus spp. (see small narcissus flies), 60 

European Union, 56, 63, 135, 137 

Eustephia yuyuensis, 244 

Exports; bulbs, 56f, 63; cut-flowers, 57 

Facial erythema, 407 

Facial neuralgia, 345 

Facial paralysis, 344f 

Fatigue syndromes, 273, 416 

Feeding experiments, 163, 165, 168 

Fenamiphos, 85 

Fertilisers and recommendations, 70ff, 263 

Fibonacci series, 7 

Field operations, 69ff, 86ff, 90f, 

Financial returns, 131 

Fire (disease), 61, 87, 89 

Flailing, 92 

Flooding, 101 

Flower; abnormalities, 13; cropping, 55, 

88; development, 11, 13, 60, 74; flower 

foods, 404; forcing (glasshouse), 2, 12, 

64, 75f, 88, 94; formation, 12; initiation, 

11, 60; numbers, 11, 91; pickers/ 

gatherers, 394, 401, 403, 405, 407; sales, 

132; stalk, 4, 13 

Flowering (anthesis), 2ff 

Flowers, 4f, 10, 13 

Fluazinam, 407 

Fluorometric methods, 291, 293ff, 299, 373 

Foliar diseases, 87 

Folk medicine, 26f 

Folklore, 19ff, 410 

Food shortage, 27 

Food use, 27 

Formaldehyde, formalin, 75f, 88, 94 

Formylgalanthamine, 290 

Formylnorbelladine, 317 

Formylnorgalanthamine, 215ff 

Fortucine, 248 

Fragrances, 410f, 418 

Freezing stress, 99 

Frost, 61 

Fumigation, 72 

Fungal diseases (plant), 61, 72, 81 

Fungal diseases (skin), 412 

Fungi, antagonistic, 72ff, 101 

Fungicides, 61, 76, 87, 91, 94ff, 134, 407 

Fusarium oxysporum (see basal rot) 

Galanthamine, 25, 141f, 171f, 186, 195, 215ff, 

334ff, 371ff, 412f; absorption, 338; 

accumulation, 217ff, 247f, 339; adamantyl 

ester, 274, 282, 350; analogues, 244, 282, 

410, 415ff; biosynthesis, 171; clinical trials, 

335, 341, 343, 357ff; content, 247; 

crystallization, 267; decomposition, 288, 

299; derivatives, 196, 244, 274, 349; 

distribution, 336; dose, 281; drug delivery, 

338, 414f; drying for extraction, 248; 

excretion, 336, 338ff; extraction, 108, 110, 

248ff, 256ff, 264, 279, 282, 286ff, 342, 394, 

408f; extraction artefacts, 178, 268; 

extraction emulsions, 287; extraction pH, 

267; extraction solvents, 266, 268, 287, 395; 

isolation, 269; measurement, 249, 293ff; 

metabolism, 339ff; precursors, 163; price, 

274; production, 54, 59, 196, 215ff, 247ff, 

256ff, 273ff, 340, 372, 413; safety data, 349; 

side-effects, 215, 346, 349f, 359, 361, 365; 

sources, 242ff, 275ff; structure, 306; supply 

and demand, 274; synthesis, 110, 196, 215, 

256, 274, 304ff, 342, 350, 367, 372, 412; 

tolerance, 343 

Galanthamine content, 244, 248, 256ff, 

286ff, 411; agronomic factors, 273ff; 

bulb development, 276; environment, 

280; fertilizers, 262; plant parts, 261; 

yearly fluctuations, 248, 269 

Galanthamine hydrobromide, 195, 242, 

249, 268, 281, 304, 337, 360, 372, 413f

Galanthamine-type alkaloids, 169ff, 179, 

184, 323 

Galanthaminone, 291f, 339ff 

Galanthine, 166, 242 

Galanthus, 25, 195, 215, 242, 244ff, 256f, 276, 

292, 304, 306, 334, 371, 410, 412f, 415 

Galathindin, 243 

Galen, 25 

Garden swift moth, 60 

Gastroenteritis, 26 

Gemanthamine, 248 

Genetic material, conservation, 31, 103 

Genetics, 31 

Geophytic habit, 2 

Gerard, John, 19 

Gibberellins, 4, 197, 221, 226 

Gladiolus, 221, 226, 229, 231 

Glaucoma, 346 

Globodera spp., 60 

Glomus spp. (see endomycorrhizal fungi) 

Glutaraldehyde, 75f 

Glycopeptides, 384, 400 

Glycoproteins, 400 

Goats, 26 

Grassiness, 61 

Gravimetric methods, 295 

Grey bulb rot, 61 

Gross margin, 131ff 

Groundkeepers (see volunteers) 

Growth regulators, 221 

Guillain-Barre syndrome, 345 

Habitats, 2 

Haemanthamine, 142, 168f, 178, 215ff, 

261, 267, 274, 277, 289f, 292, 299 

Haemanthamine-type alkaloids, 168ff, 178 

Haemanthidine, 168 

Haemanthus katherinas, 245 

Haemodoraceae, 276 

Hair shampoos, 411 

Hallucinogenic effects, 184 

Harmful effects of narcissus, 185, 401ff 

Hepialus lupulinus (see garden swift moth) 

Herbal medicines, 24ff, 185, 345 

Herbal, Culpeper’s, 22, 25f 

Herbal, Gerard’s, 19 

Herbal, John K’Eogh’s Irish, 25 

Herbicides, 68, 86ff, 90, 92, 101 

Herrick, Robert, 24 

High-health status bulbs, 88 

Hipotensive, 185 

Hippeastrum, 245, 390 

Hippocrates of Cos, 25 

Histiostoma species, 60 

Homer’s Odyssey, 334 

Homoeopathic medicine, 25 

Homolycorine, 142, 167, 406 

Index 423 

Homolycorine-type alkaloids, 163, 167, 

177, 180 

Hoop Petticoat daffodils, 45 

Hordenine, 292, 295 

Hormones, 2, 196 

Horticultural classification, 47ff 

Hot-water treatment, 60, 64, 72–88, 134, 139 

Hydrochloric acid, 77 

Hydrogen fluoride, 97 

Hydrogen peroxide, 76 

Hydroxyvittatine, 169 

Hygiene, 72 

Hymenocallis, 245, 295 

Hypobaric storage, 99 

Hypoxidaceae, 276 

Hysteranthy, 4 

Hysterical affections, 26 

Immunoassays, 288, 293 

Imports, bulbs, 56f 

Incartine, 166 

Indolylacetic acid, 77, 197 

Infantile paralysis, 410 

Infrastructure, 69 

Initiation; bulb unit, 8, 10ff, 14; floral, 7ff 

Insecticides, 76ff, 85ff, 196 

Insects, 13, 389 

Integrated Crop Management, 100ff 

International Registration Authority, 50, 62 

Intraocular pressure, 195, 346 

Ion-exchange resins, 413 

Iris, 2, 11 

Iron, 71 

Irrigation, 65, 89f 

Irritancy, 185, 405 

Ismene (Hymenocallis), 170, 245 

Isofenphos, 78 

Isolectins, 384ff 

Isomers, 306, 411 

Ixiolirion, 245f 

Jet-lag, 273, 417 

Joint disorders, 416 

Jonquil absolute, 185 

Jonquils, jonquilla cultivars, 27, 39, 59, 74, 

100, 394 

Juvenile phase, 14 

Kinetin, 218ff 

Kohut, Heinz, 23 

Labour, 56, 69, 131, 135, 137 

Lamium album, 418 

Language of flowers, 20f 

Large narcissus fly, 60, 73, 76, 78, 81, 85, 88, 91 

Large-cup cultivars, 1, 47, 49, 59, 74, 257, 269 

Leaf cuttings (propagation), 106

424 Index 

Leaf numbers, 11 

Leaf scorch, 61, 87, 97 

Leaf senescence, 2, 88f, 91 

Leaves and leaf bases, 4ff, 11f 

Lectin content; life cycle effects, 389; 

varietal differences, 383 

Lectins, 185, 382ff, 389f, 411; applications, 390; 

carbohydrate-binding, 388; classification, 

382; concentration, 385; isolation, 382; 

mannose-binding, 383, 390; mitogenic 

activity, 389; molecular cloning, 385; 

purification, 382, 384; role in plants, 389f; 

snowdrop, 385f, 390; structure, 384, 386 

Legends, 22ff 

Leucojum, 170, 196, 215, 242ff, 247, 249f, 

256f, 274, 276, 291ff, 304, 372, 413 

Leukaemia, 351 

Lewis lung carcinoma, 351 

Light (effects of), 11 

Liliaceae, 390 

Lilium, 231 

Lily rash, 401 

Linseed, 134f 

Lithium, 346 

Liver function, 340, 349 

Longevity (life span), 5, 7 

Longidorus spp., 60 

L-selectride, 319, 323 

Lycoramine, 277, 282, 288, 292, 304, 306, 

325, 327ff, 414 

Lycorenine, 167, 288f, 292, 294 

Lycoricidine, 274 

Lycorine, 26, 141f, 165ff, 177, 180, 186, 

196, 243ff, 249, 274, 277, 287ff, 413 

Lycorine hydrochloride, 243 

Lycorine synthesis, 196 

Lycorine-type alkaloids, 164ff, 177, 180 

Lycoris, 166, 245, 288, 413 

Magnesium, 70f 

Magnicoronati narcissus, 47 

Male impotence, 273 

Mammals, 184 

Mandrax, 345 

Manganese, 70 

Mania, 344, 416 

Masonin, 405ff 

Mass spectrometry, 171, 180ff, 291, 299 

Mediicoronati narcissus, 47 

Meristematic tissues, 277 

Meristem-tip culture, 104 

Merodon equestris (see large narcissus fly) 

Mesembrane- (Sceletium-) type alkaloid, 141 

Mesembrenone, 141 

Metam-sodium, 70 

Methyl bromide, 72 

Methylapogalanthamine, 269 

Methylgalanthamine hydroxide, 244 

Methylnorbelladine, 165ff, 320 

Methyloduline, 258 

Metrifonate, 359 

Microbiological reduction, 314 

Micropropagation (see tissue culture) 

Migraine, 345 

Mineral oil sprays, 88 

Miniature daffodils, 27 

Mites, 73 

Mn (III) acetylacetonate, 314, 320 

Mononucleosis, 416 

Montanine, 142, 169 

Montanine-type alkaloids, 167ff, 183 

Morphine, 342 

Mother bulbs, 5, 7 

Mucilage, 185, 405 

Mucor plumbeus, 75 

Mulches, 85 

Multi-headed cultivars, 1 

Multi-nosed bulbs, 7 

Multiple sclerosis, 344 

Muscarinic agonist, 414 

Muscle function, 336, 417 

Muscular dystrophy, 242, 345 

Muscular relaxant antagonists, 242 

Myalgic encephalitis, 416 

Myasthenia, 195, 242 

Mycoplasma-like organisms, 61 

Mycorrhizae, 14 

Myopathy, 242 

Mythology, 22ff 

Narciclasine, 25, 142, 169f, 186, 197, 274, 

295, 408 

Narciclasine biosynthesis, 169 

Narciclasine-type alkaloids, 167ff, 178 

Narcissidine, 166 

Narcissin, 243 

Narcissine, 26 

Narcissistic personality disorder, 23 

Narcissimum (ointment), 26 

Narcissus; anatomy, 10, 12; annual growth 

cycle, 2ff, 59f, 103; constituents, 399ff; 

distribution, 1, 31, 242; extracts, 185; 

fragrance, 408; genes, 103; growth/ 

development, 4ff, 277; production for 

medicine, 276; species and cultivars, 1f, 

100, 106, 142ff, 276 

Narcissus (mythological character), 22 

Narcissus absolute, 397, 400, 411 

Narcissus leaf miner, 60 

Narcissus oil, 25f 

Narcissus-type radiation effect, 409, 411 

Narcotic, 26 

Narwedine, 243, 282, 296, 304, 306, 320f, 

329, 413

Narwedine derivatives, 414 

Narwedine-type enone, 325f 

Naturalising bulbs, 65 

Neck rot (pathological), 61, 92 

Neck rot (physiological), 61 

Nectria radicicola, 60, 100 

Nematicides, 85, 88 

Nematodes, 60f, 70ff, 88, 91, 100ff, 411 

Neostigmine, 256, 336, 342, 344f, 338 

Nephrolepis, 226 

Nerine, 167, 221, 226, 245 

Nervous system diseases, 414 

Neuritis, 242 

Neurological effects, 336, 344f, 415 

Neuromuscular diseases, 195 

Neuropsychological function, 364 

Nicotine dependence, 274, 417 

Nicotinic agonist, 410, 415 

Nitrogen, 70f 

Nivalidine, 292, 294 

Nivalin(e), 244, 249, 304, 334, 345, 371 

Nivaline Jikon, 242 

Norbelladine, 142, 163, 170, 305, 325 

Norellia spinipes (see narcissus leaf miner), 60 

Norgalanthamine, 171, 304, 307, 312, 325, 

414, 416 

Norpluviine, 164ff, 258 

Nuclear magnetic resonance spectroscopy, 

172, 177, 179f, 269, 288 

Numbness, 26 

Obesine, 179 

Offsets, 7 

Oilseeds, 134f 

Omethoate, 88 

One-year-down growing (see crop duration) 

Onions, 134, 218, 231 

Opiates, 256 

Optimised field production, 103, 108, 277 

Orchidaceae, 390 

Organic fertilizers, 71 

Organic production, 100ff 

Organogenic tissue, 216 

Ornamental horticulture (classical times), 246 

Ornamentals, 1, 53, 80, 242, 279 

Osteoarthritis, 416 

Oxamyl, 88 

Oxocrinine, 169 

Paclitaxel, 371 

Paclobutrazol, 71, 99, 221ff 

Palsy, 26 

Pancratine, 295 

Pancratistatin, 286, 295 

Pancratium, 170, 244f, 290, 292f 

Pancuronium-induced neuromuscular 

blockade, 338 

Index 425 

Panic disorders, 417 

Paper electrophoresis, 294 

Paperwhite narcissus (paper whites), 41, 97, 

99, 405 

Papillon daffodils, 47, 49 

Paralysis, 26, 344 

Paratrichodorus spp., 60 

Parkinson’s disease, 367, 417 

Parvicoronati narcissus, 47 

Patents, 410ff; extraction, 412; formulation, 

414f; non-therepeutic use, 410f; 

pharmaceutical, 410ff; production, 412; 

therapeutic use, 415 

Penicillium bulb rots, 61, 75f, 100 

Perfume, perfumery, 394ff, 411 

Perianth segments (see tepals) 

Peripheral neuropathy, 344 

Peroxyacetic acid (peracetic acid), 76 

Personality disorders, 23 

Pest avoidance, 91 

Pest forecasting models, 88 

Pesticide applications, 67, 85, 100 

Pesticide run-off, 67 

Pesticide stability, 77 

Pesticides, 280, 405 

Pests (in general), 60ff, 83 

Phaedranassa megistophylla, 276 

Pharmaceuticals, 141, 256, 276, 279f 

Pharmacokinetics, 332ff 

Pharmacology, 184ff, 243, 334ff 

Pheasant’s eye cultivars, 34 

Phenanthridine alkaloids, 26 

Phenolic coupling, 220, 313, 320f, 325, 411 

Phenolic oxidation, 320, 325 

Pheromone, 185 

Philodendron, 226 

Phorate, 88 

Phosphate, 70f 

Photometric methods, 249 

Photoperiod, 9 

Physiological disorders, 13, 61 

Physiotherapy, 340 

Physostigmine, 195, 215, 256, 336, 341, 

346, 359, 373 

Picloram, 216 

Pigs, 26 

Plant growth regulators, 88, 185 

Plant health authorities, 63, 90, 98 

Plant introductions, 31 

Plant morphology, 4ff 

Plant Passports, 63 

Planthopper, 389 

Plasma concentrations, 338 

Pliny, 21, 25 

Pluviine, 166f 

Poetaz cultivars, 2 

Poeticus cultivars, 74, 77

426 Index 

Poet’s cultivars, 34 

Poison, poisoning, 26f, 184, 242, 345, 417 

Polarographic methods, 249, 295 

Poliomyelitis, 242, 256, 413 

Pollen, 13, 102 

Poly/dermatomyositis, 344 

Polysaccharides, 4 

Popularity (of flower crops), 21, 53 

Post-polio paralysis, 195, 344 

Potash, 70 

Potassium, 71f 

Potato cyst nematode, 60, 69f 

Potatoes, 134ff, 226 

Pot-grown narcissus, 57 

Poultices, 345 

Poultry-keepers, 23 

Powelline, 295 

Pratylenchus penetrans, 60, 70, 100f 

Pre-cooling, 79 

Predation, 2 

Predictive crop models, 79 

Pre-eclampsia, 416 

Pre-soaking, 73, 77 

Pretazettine, 26, 168, 178, 186, 195, 274, 

306, 350f 

Pretazettine synthesis, 197 

Pre-warming, 78 

Primordia, 10 

Probes, 390 

Prochloraz, 76 

Producing countries, 55, 281 

Profitability of bulb growing, 131ff 

Progressive myodystrophy, 344 

Propagation, 10, 102ff, 278 

Prophage induction, 185 

Protective adaptation, 242 

Protective clothing, 405 

Protein crops, 135 

Protein synthesis, 196, 274, 351f 

Proteolytic diseases, 416 

Pseudo-botanic names, 47 

Pseudonarcissus group, 30 

Psoriasis, 416 

Psychometric and neuropsychiatric tests, 361 

Pyrethroids, 88 

Pyridostigmine, 336, 338 

Quantitative analysis, 286ff 

Quarantine pest, 63 

Quercetin, 26 

Radiometric techniques, 293, 372 

Rainfall, 65 

Ramularia vallisumbrosae (see white mould) 

Raphides, 12f, 405 

Rapid screening, 374 

Rash, 402, 405 

Recovery rate, 342 

Red List, 1, 246 

Register, 51 

Register and Classified List, 51f 

Religious connections, 23f 

REM sleep, 349 

Repetative strain injury, 404 

Resistance to fungicides, 87 

Respiration, 243, 345, 348 

Respiratory diseases, 25 

Retarding flowering, 98ff 

Retention of learned behaviour, 343 

Retrovirus infection, 390 

Reversal of paralysis, 342, 344, 371 

Rheumatoid arthritis, 416 

Rhizoctonia tuliparum, 61 

Rhizoglyphus spp., 60 

Rhizopus spp., 61, 97 

Rhodophiala bifida, 169 

Riddles, 94, 97 

Ridge- and bed-growing, 79f, 84 

Rivastigmine, 357 

Roguing, 90 

Root; cap, 13; cultures, 215; hairs, 13f 

Root rot, 60f, 100 

Root-lesion nematode, 60 

Roots, 13ff, 72; adventitious, 4, 14; 

contractile, 14 

Rose, 406 

Rosellinia necatrix, 61 

Royal Horticultural Society, 47, 49, 50, 62 

Rust, 85 

Sanguinine, 304, 324f, 414 

Sap, 2, 16, 401ff 

Scales (bulb scales), 4f, 10, 14 

Scape (see stem) 

Scent, 185 

Schiff’s bases, 163 

Schizophrenia, 243, 273, 346, 417 

Sclerotinia bulborum, 61 

Sclerotinia polyblastis, 61 

Scooping (propagation), 105 

Scopolamine, 195, 336f, 343, 345, 358 

Scopolia maxima, 225 

Seasonal workers, 404 

Secondary metabolism, 215, 219 

Section (taxonomic), 30; Apodanthi, 49; Aurelia, 

44; Braxireon, 41; Bulbocodium, 45, 49; 

Chloraster, 40; Ganymedes, 30, 38; Helena, 34; 

Hermione, 30, 41; Jonquilla, 49; Jonquillae, 38; 

Narcissus, 34; Pseudonarcissus, 35; 

Tapeinanthus, 41; Tazettae, 41, 49 

Securinine, 294 

Seed germination, 14 

Seedling growth, 408 

Seedlings, 14ff

Seeds, 13, 109, 185, 197, 215f, 232, 234 

Selection, 90 

Series (taxonomic); Albiflorae, 42; 

Hermione, 42 

Serum levels, 338 

Sesamum indicum, 418 

Set-aside, 135 

Sewage sludge, 71 

Sexual dysfunction, 418 

Shakespeare, William, 23 

Sheet-laminated fabric, 411 

Shoot dormancy, 232 

Shoot-clump cultures, 104, 215ff, 248 

Shoots, adventitious, 221, 231 

Single Market, 56 

Single-nosed round bulb, 4, 7 

Site considerations, 64ff 

Skin (human), 407, 413, 418 

Skin diseases (bulbs), 61, 100 

Slime vessels, 405 

Slugs, 61 

Small narcissus flies, 60 

Small-cup cultivars, 1, 47, 49, 59, 74 

Smoke treatments, 12, 100 

Smoulder, 61, 87, 89, 100 

Snowdrop (see Galanthus) 

Socrates, 26 

Soft rot, 61, 97 

Soil, 68; compaction, 68; disinfection, 70, 

88, 101; erosion, 86; moisture, 89; 

removal, 94; sampling, 70 

Soil-borne fungi, 70 

Solar sterilization, 101 

Somatic embryo, 105, 217, 220 

Spastic pareses, 242 

Spathe, 13 

Species; N. abscissus, 36; N. albescens, 36; 

N. albimarginatus, 40; N. alpestris, 36f; 

N. angustifolius, 246; N. assoanus, 39, 43; 

N. asturiensis, 35f; N. atlanticus, 40; 

N. aureus, 42; N. baeticus, 39; N. barlae, 43; 

N. bertolonii, 42; N. bicolor, 36; N. blancoi, 

45; N. broussonetti, 44; N. bujei, 36; 

N. bulbocodium, 45f, 105, 109, 185; 

N. calcicarpetanus, 36; N. calcicola, 27, 40; 

N. canaliculatus, 42; N. canariensis, 43; 

N. cantabricus, 45f; N. cavanillesii, 41; 

N. cerrolazae, 39; N. confusus, 36, 196, 

216ff, 248f, 290; N. corcyrensis, 42; 

N. cordubensis, 39; N. cuatrecasasii, 40; 

N. cupularis, 42; N. cyclamineus, 27, 31, 36, 

49, 109; N. cypri, 42; N. dubius, 43; 

N. elegans, 4, 31, 43ff; N. fernandesii, 39; 

N. fontqueri, 36; N. gaditanus, 39f; N. gayi, 

37; N. genesii-lopezii, 37; N. gracillis, 245; 

N. graellsii, 46; N. hedraeanthus, 45f; 

N. hispanicus, 36f, 53; N. humilis, 4; 

Index 427 

N. incomparabilis, 166; 245; 

N. italicus, 42; N. jacetanus, 36; 

N. jacquemoudii, 46; N. jeanmonodii, 45; 

N. jonquilla, 12, 38f, 102, 184, 245, 394; 

N. juncifolius, 12, 39; N. juressianus, 45; 

N. lagoi, 36; N. lainzii, 45; N. lobularis, 13, 

245; N. longispathus, 13, 37; 

N. lusitanicus, 38; N. macrolobus, 37; 

N. marianicus, 39; N. minor, 36; 

N. minutiflorus, 39; N. moleroi, 37; 

N. moschatus, 37; N. nanus, 35f; 

N. nevadensis, 37; N. nobilis, 36f; N. obesus, 

45f; N. obvallaris, 37; N. ochroleucus, 42; 

N. pachybolbos, 43; N. palearensis, 39; 

N. pallens, 39; N. pallidiflorus, 37; 

N. pallidulus, 38, 141; N. panizzianus, 43; 

N. papyraceus, 41ff, 105, 184; N. parviflorus, 

36; N. patulus, 42; N. perez-chiscanoi, 37; 

N. peroccidentalis, 46; N. poeticus, 1, 14, 25, 

27, 34f, 49, 71, 184, 245, 249, 394; 

N. polyanthus, 43; N. portensis, 36; 

N. primigenius, 37; N. provincialis, 36; 

N. pseudonarcissus, 1f, 7, 13, 25, 31, 35ff, 

109, 184f, 245, 411; N. pumilus, 36; 

N. radiiflorus, 34f; N. radinganorum, 37; 

N. requienii, 39; N. romieuxii, 33, 45ff; 

N. rupicola, 40; N. scaberulus, 40; N. serotinus, 

4, 31, 44; N. subnivalis, 45; N. tazetta, 1, 4, 

12ff, 23, 26, 31, 41ff, 71, 73, 88, 245, 289, 

350, 418; N. tingitanus, 47; N. tortifolius, 

43; N. tortuosus, 37; N. triandrus, 13, 27, 

38, 49, 109, 245; N. undulata, 245; 

N. viridiflorus, 4, 31, 40; N. watieri, 27; 

N. willkommii, 39 

Species concept, 30 

Spectrophotometry, 172ff, 249, 258, 289, 

293f, 372 

Spinal impotence, 242 

Split-corona cultivars, 47, 49 

Sprekelia formosissima, 167, 168 

Stages of development, 389 

Stagonospora curtisii, 61 

Starch, 13 

Statistics (bulb growing), 53ff 

Stem, 13 

Stem apex, 10ff 

Stem extension, 2, 12, 411 

Stem nematode, 63, 68, 68–91 

Steneotarsonemus laticeps, 60 

Stereochemistry, 176, 177, 182 

Sterilants, 70 

Sternbergia, 244f, 289, 295 

Storage proteins, 389 

Stories, 23 

Straw, 85, 101 

Stress, 417 

Structure–activity relationships, 244

428 Index 

Subgenus (taxonomy); Ajax, 35; 

Angustifolii, 43; Apodanthi, 39; 

Chloranthi, 40; Corbularia, 34, 44; 

Eu-narcissus, 34; Hermione, 34, 41; 

Jonquillae, 38; Narcissus, 34; 

Serotini, 44 

Sucrose, 218ff 

Sugar beet, 134ff 

Suicide, 184 

Sulphuric acid, 92, 268 

Summer daffodil crops, 100 

Sun scorch, 61 

Supercritical fluid extraction, 287 

Suspension cultures, 216 

Sustained release composition, 415 

Symptoms, 184 

Tacrine, 195, 215, 336, 341, 342, 359, 415 

Tagetes, 102 

Tapeinanthus, 30, 41 

Taxonomy, 1, 30ff, 143 

Tazetta narcissus, 4, 12, 30, 41, 54–65, 79, 

84, 99ff, 403, 405 

Tazettine, 142, 179, 182, 215ff, 288ff 

Tazettine-type alkaloids, 167ff, 178, 182 

Temperature (effect on narcissus), 9, 11ff, 

65, 79, 98 

Tepals, 13 

Terminal bud, 4 

Tetraploids, 102 

Texts, 2 

Thebaine, 409 

Thiabendazole, 76, 77, 94 

Thiram, 76 

Tinea pedis, 410 

Tissue culture, 105, 196, 219, 227ff, 240, 

290, 411 

Tissue culture, liquid media, 96, 216ff, 226 

Tissue culture, seed-derived, 221 

Tissue differentiation, 220 

Topography, 65ff 

Tortoise, 27 

Tourette’s syndrome, 417 

Tourism, 56 

Toxicology of galanthamine, 27, 184f, 

334ff, 347, 407 

Trance, 184 

Transport of bulbs, 98, 99 

Trasformation, 103 

Triandrus narcissus, 30, 59, 100 

Trichodorus spp., 60 

Trimethylsilyl derivatives, 288 

Tropic response, 13 

Trumpet cultivars, 1, 47, 49, 59, 74, 405 

Tubocurarine, 195 

Tulips, 2, 5, 10f, 69, 231, 408 

Tumors, 25f 

Tunic (bulb skin), 5 

Twin-scaling (propagation), 105ff, 220, 

233, 278 

Two-year-down growing (see crop duration) 

Ulcerative colitis, 414 

Ungernia, 244ff, 256, 295, 306, 413 

Ungiminorine, 245, 377 

Uniconazole, 99 

Urginea indica, 215, 219 

UV, MS and biochemical detection, 374 

Vallota purpurea, 245 

Variable costs, 132 

Variable inputs, 131 

Vase-life, 408 

Vegetable cropping, 137 

Velnacrine, 359 

Vertebrate pests, 61 

Victorians, 20 

Virus, Epstein Barr, 416 

Virus, herpes simplex, 243 

Virus, human Immunodeficiency, 389, 

412, 416 

Virus, human T lymphotropic, 412 

Viruses, 286, 351 

Viruses (plant), 61, 63, 70, 88, 90f 

Virus-tested plants, 63, 88, 104 

Vitrification, 221ff 

Vittatine, 168ff, 292, 294 

Volume of distribution, 338f 

Volunteers, 69, 87 

Water table, 89 

Waterlogging, 61, 65, 90 

Weather (effect of), 65 

Weed competition, 86, 280 

Weeds (see herbicides) 

Western Europe, 53 

Wheat, 134 

White mould, 61, 87 

White root rot, 61 

Wild-collected bulbs, 1, 100, 276 

Willgerodt-Kindler reaction, 318 

Wind, 65 

Windrowing (drying in field), 93 

Wordsworth, William, 19 

Xiphinema diversicaudatum, 60 

Zephyranthes, 163, 215, 245 

Zinc, 71 

Zineb/maneb, 76

Narcissus and Daffodil

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