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Journal of Scientific and Engineering Research, 2017, 4(9):351-355

ISSN: 2394-2630
Research Article CODEN(USA): JSERBR

Qualitative phytochemical screening and antimicrobial activity of methanol extract of

Maesobotrya barteri (Baill.) Hutch

Uwemedimo E. Udo*, Nyakno E. Akan and Enwongo M. Udoh

Department of Chemistry, Faculty of Science, University of Uyo, P.M.B. 1017, Nigeria

Abstract The stem bark extract of Maesobotrya barteri was screened for antimicrobial activity against some
pure cultures of bacterial and fungal species. These were carried out by the Plate -hole diffusion method on
Mueller – Hinton agar (MHA) for bacteria and Sabouraud Dextrose Agar (SDA) for the fungi. The Minimum
Inhibitory Concentrations (MICs) of test samples found to be active by the diffusion test were determined based
on the macro dilution method. The crude extract showed activity against Staphylococcus aureus, Shigella
dysenteriae, Salmonella typhi, Microsporum spp and Candida albicans. Qualitative phytochemical screening
revealed the presence of alkaloids, flavonoids, tannins, terpenes, saponins and cardiac glycosides. This result
confirms its ethnomedicinal use in the treatment of microbial infections.

Keywords Antibacterial, bioactive compounds, Maesobotrya barteri, medicinal plants

Introduction
Maesobotrya barteri(bush cherry) is a dioecious tree or shrub belonging to the family Euphorbiaceae with a
simple indumentum up to 10 m high. It is a rainforest plant occurring in Sierra Leone, Southern Nigeria,
Western Cameroun and Congo Basin [1]. The leaves are alternate, often long-petiolate, stipulate simple, entire
or toothed and penninerved. The wood is light and burnt for fuel [2]. Maesobotrya barteri bears fruits from
April to June, which is up to 1cm long ovoid, and often distinctly pointed [3].
In Nigeria, the plant is known by many local names such as “Oruru” (Benin), “Olowunor Obomodu” (Yoruba),
“MiriỌgu” (Igbo) and “Nyanyated” (Ibibio). The fruits are succulent black-purple berries. They are edible and
stain the tongue. The seed are often with a conspicuous caruncle with the endosperm present or absent. In Sierra
Leone, perhaps due to superstition, the plant is used to make cages in dwellings for twins although the
significance is not explained. Stem bark of Maesobotrya barteri var. sparsiflora and Trichilia monadepha stem
bark are boiled together and the decoction is taken for abortion [4]. A bark decoction of Maesobotrya barteri is
taken in Congo (Brazzaville) for dysentery, urethral discharge and as an aphrodisiac [2]. For a very longtime, M.
barteri has been in use in the local communities for the treatment of diarrhea, stomachache, dysentery, urethral
discharge, venereal disease, jaundice, cough and others [3]. The twigs and small branches are used as chewing
stick [5]. Some are eaten as a refreshing delicacy and the fruits serve as a potential source of raw material for
juice making that can support the economic and industrial development in Nigeria. Also the fruits enhance
frequent waste elimination, including acid, sterols and fat [6]. The fiber is of benefit in diverse diseases [7] and
helps lower cholesterol absorption by preventing the formation of plaques [8].
Studies on Maesobotrya barteri have shown its major, minor and trace element composition as well as its wide
use as chewing sticks in Southern Nigeria [9] and its fruit nutritive value [10]. The use of crushed root and sap
of M. barteri on the skin externally has been reported to be effective in treating some skin infections [11]. The
antimicrobial potential of the crude extract of aerial plant parts of M. barteri as well as the corrosion inhibition
ability of its leafextract in acid solution have also been reported [12, 13].

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To our knowledge, the in vitro antimicrobial activity of methanol stem bark extract of Maesobotrya barteri has
not been reported despite its widespread medicinal use. In the present study, the phytochemical and in-vitro
antimicrobial activity of stem bark extract of Maesobotrya barteri is presented using standard analytical
methods.

Material and Methods

Plant materials: The fresh stem barks of Maesobotrya barteri (bush cherry) were collected from a bush in Oruk
Anam Local Government Area of Akwa Ibom State, Nigeria in the month of June, 2016. The time of collection
coincided with the rainy season in Nigeria. Plant identification, authentication and specimen referencing were
done at the Department of Botany and Ecological Studies in the Faculty of Science, University of Uyo, Nigeria.
Sample preparation: The stem barks of M. barteri were thoroughly washed with distilled water to remove any
trace of dirt sticking to the surface of the stem barks. The stem barks were chopped into small pieces and air
dried for 7 days. The particle sizes were further reduced by grinding using a Thomas Wiley machine and stored
in an air-tight plastic container, properly labeled prior to analyses.
Extraction procedure: The dried pulverized stem barks (500 g) were thoroughly macerated with 99.5 %
methanol (5 L) for 7 days at room temperature. The sample mixture was filtered and the filtrate concentrated to
dryness in vacuo at 40 oC using a rotary evaporator to afford a brown colored extract. The extracts were stored
in a sealed container and kept in a refrigerator at -4 °C until use. All reagents and chemicals used in this work
were of analytical (AnalaR) grade and were sourced from Sigma-Aldrich chemical company, United Kingdom.

Qualitative determination of phytoconstituents: Qualitative tests to identify the constituents of the extract were
performed using standard procedures outlined elsewhere [14, 15]. Precisely, screening of alkaloids was carried
using Dragendroff’s and Mayer’s reagents, saponins by Frothing and Fehling’s tests. Cardiac glycosides were
detected by Liebermann’s and Keller-Killiani’s tests, tannins by the Ferric chloride test and phlobatannins by
hydrochloric acid test. Flavonoids were detected by the magnesium metal/hydrochloric acid test, triterpenes by
the chloroform/acetic anhydride/sulfuric acid test and anthraquinones by the benzene/ammonia solution test.

Microorganisms
The bacterial test strains used in this study were Staphylococcus aureus, Klesbsiella spp, Pseudomonas
aeroginosa, Shigella dysenteriae, Salmonella typhi, Esherichia coli, Microsporum spp, Trichophyton spp,
Epidermophyton spp, Aspergillus flavus and Candida albicans. These clinical samples were obtained from St.
Luke’s Hospital Anua and Microbiology laboratory of University of Uyo, Nigeria. Fungal isolates were
collected from the environment and Dermatophyte from some school children all in Uyo Metropolis, Nigeria.
The isolates were purified by sub-culturing into their selective medium and thereafter sub-cultured into nutrient
agar.
Evaluation of antibacterial and antifungal activities of the extract
Plate-hole diffusion test: The evaluation of antimicrobial activity of the stem bark extract of Maesobotrya
barteri was carried out by the Plate–hole diffusion method [16] on Mueller-Hinton agar (MHA) for bacteria and
Sabouraud Dextrose Agar (SDA) for the fungi. Solutions of the extract and fractions were prepared in 10%
Tween 80 to concentrations of 100, 50, 25 and 12.5 mg/ml. The innocula of the microorganisms were prepared
separately from 12 h broth cultures (Mueller-Hinton broth for bacteria and the Sabouraud dextrose broth for the
fungi) and incubated at 37 °C. All culture media and distilled water were sterilized at 121°C for15minutes in an
autoclave. These innocula were diluted with sterilized distilled water to obtain a density corresponding
approximately to 0.5 of McFarland standard turbidity scale (108) colony forming unit “CFU” per mlfor the
bacteria and 103 spores per ml for fungi) [16]. Each innoculum (0.5 ml) was introduced into the corresponding
fluid agar medium homogenized and 25 ml of it poured into sterile plastic petri dishes. The petri dishes were
allowed on the flat slab top for the medium to solidify within 30 min. A standard cork borer of 5mm in diameter
was used to cut four equidistant uniform wells per plate on the surface of different plates into which was added
50µl solution of the extract at varying concentrations 12.5, 25, 50 and100 mg/ml.

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The reference drugs were Gentamicin, batch 20070402 (0.4mg/ml) and Nystatin batch 04D05 (500µg/ml). The
plates were incubated at 37 °C for 24 and 48 h for the bacteria and fungi respectively. The antimicrobial activity
was evaluated by measuring the zone of inhibition around the hole. Each test concentration had three
replications. The results were recorded as the mean diameter of the zones of growth inhibition surrounding the
discs [17]

Determination of minimum inhibitory concentrations (MIC) using macrodilution method

The Minimum Inhibitory Concentrations (MICs) of test samples found to be active by the diffusion test were
determined based on the macrodilution method [16] with some modifications as follows. The test
extract/fractions were dissolved in 10% Tween 80 to give a stock concentration of 100 mg/ml and serially
diluted (two-fold) in a series of test tubes to a working concentration ranging from 1.560 to 100 mg/ml using
nutrient broth supplemented with 10% glucose and 0.05% phenol red (colour indicator). These were later
inoculated with 0.2ml suspension of the test organisms. Microbial growth was determined by observing for
color change in the tube (red to yellow when there is growth). The lowest concentration that showed no change
of color was considered as the MIC.
Table 1: Phytochemical composition of methanol stem bark extract of Maesobotrya barteri
Phytoconstituents Detection
Alkaloids Present
Tannins Present
Terpenes Present
Cardiac glycosides Present
Flavonoids Present
Saponins Present
Anthraquinones Absent
Phlobatannins Absent
Deoxy sugar Absent

Table 2: Inhibition zone diameter, IZD (mm) of methanol extracts of Maesobotrya barteri on some test
organisms
Test Organisms Zones of inhibition (mm) with different Gentamycin Nystatin500
concentrations of methanol extract 0.4 mg/ml µg/ml
100 mg/ml 50 mg/ml 25 mg/ml 12.5 mg/ml
ME ME ME ME
S. aureus 18 14 12 10 23 -
Klebsiellaspp - - - - 16 -
P. aeroginosa - - - - 12 -
S. dysenteriae 15 12 10 8 7 -
S. typhi 10 8 6 6 7 -
E. coli - - - - 19 -
Microsporumspp 10 - - - - 15
Trichophytonspp - - - - - 22
Epidermophytonspp - - - - - 29
A.flavus - - - - - 20
C. albicans 20 17 14 12 - 24

Table 3: Result of minimum inhibitory concentration (MIC) of stemb ark extract of M. barteri
Test Organisms Extract Concentration (mg/ml)
Methanol
S. aureus 6.25
S. dysenteriae 6.25
S. typhi 7.25
Microsporum spp 55
C. albicans 3.25

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Results
Phytochemical screening
The result of the phytochemical screening of the methanol extract of stem bark of M. barteri is shown in Table
1. The result shows the presence of alkaloids, flavonoids, tannins, terpenes, saponins and cardiac glycosides by
qualitative methods. Anthraquinones, phlobatannins and deoxy sugar were found to be absent.
Antimicrobial activity
Table 2 shows the diameters of the zones of inhibition exhibited by the crude extracts at various concentrations
employed. The zones of inhibition of methanol extract of M. barteri showed remarkable activities against five of
the eleven organisms tested. The zones of inhibition were compared with these standard drugs; gentamycin and
Nystatin. The crude extract showed activity against Staph. aureus, a gram-positive bacteria, S. dysenteriae and
S.typhi which are gram-negative bacteria. The crude extract was active against only two of the five fungal
species tested namely; Microsporum spp and C. albicans.
The results of minimum inhibitory concentrations (MIC) of the crude extract are shown in Table 3. The lowest
MIC (3.25 mg/ml) for the methanol extract of M. barteri was recorded against Candida albicans while the
highest MIC (55 mg/ml) was recorded against Microsporum spp.

Discussion
The phytochemical screening of the stem bark extract shows the presence of alkaloids, flavonoids, tannins,
terpenes, saponins and cardiac glycosides by qualitative methods. Anthraquinones, phlobatannins and deoxy
sugar were found to be absent in the crude extract.
The medicinal properties of these secondary metabolites are quite numerous and have been well documented
elsewhere [18-22]. The presence of these bioactive compounds in the leaves of M. barteri corroborates the
various pharmacological activities of this plant and supports its widespread use in traditional medicine.
Three of the bacterial strains tested (Gram-positive and Gram-negative were sensitive to the methanol stem
barke xtract of Maesobotrya barteri. This result is in agreement with an earlier report [12] on the activity of the
triterpene β-Amyrin from the aerial parts of Maesobotrya barteri against Staphylococcus aureus, Shigella
dysenteriae and Salmonella typhi. In contrast to this report, the stembark extract of M. barteri did not show any
activity against Klebsiellaspp, Escherichia coli and Trichophyton spp. In a similar comparison for anti-fungal
activity, the result of this study agrees in part with an earlier report [12] which showed activity of the triterpene
β-Amyrin from the aerial parts of Maesobotrya barteri against Microsporum spp but disagrees because the
crude methanol extract of the stembark of M. barteri also shows activity against the fungus Candida albicans.
The extract has fungistatic activity on only Candida albicans of all the fungi tested probably due to the
morphological differences existing between the yeast-like Candida and the other filamentous fungi. The MICs
obtained varied from 3.25 to 55 mg/ml for the crude extract of the stem bark of M. barteri. The MIC values and
the antimicrobial spectrum of crude extract of the stem bark of M. barteri indicate a remarkable antimicrobial
potency which can be exploited as promising antimicrobial agent. It is observed that the commercially perfected
antibiotics have larger zones of inhibition, indicating that drugs should be used in high purity and commercially
perfected forms. Similar trends were observed for the antimicrobial activity of methanol extract leaf extract of
guava [23].

Conclusion
In conclusion, the results of this study show that the stem bark extracts of Maesobotrya barteri possess
significant antimicrobial activity which justifies its use in traditional medicine in the treatment of microbial
infection. The bioactive compounds present in this extract may in part be responsible for the antimicrobial
activity observed in this study. Therefore, it will be interesting if the bioactive compounds are isolated and
characterized for future beneficial use.

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