Available online at www.jpsscientificpublications.com Volume – 2; Issue - 3; Year – 2016; Page: 615 – 619 DOI: 10.21276/iajmr.2016.2.3.5 Indo – Asian Journal of Multidisciplinary Research (IAJMR) ISSN: 2454-1370 PRELIMINARY PHYTOCHEMICAL SCREENING AND GC-MS ANALYSIS OF METHANOLIC LEAF EXTRACT OF Manilkara roxburghiana (Wight) DUBARD FROM SILAMBUR SACRED GROVE, TAMILNADU J. Deepa and K. Saravanakumar*, Department of Botany, Annamalai University, Annamalainagar – 608 002, Tamilnadu, India. Abstract A valuable Indian ethnomedicinal plant, Manilkara roxburghiana (Wight) Dubard was investigated in the present study to determine the phytochemical constituents present in various extracts of the leaves through GC-MS (Gas Chromatography-Mass Spectrometry) analysis. Powdered leaf plant materials were subjected to successive extraction with organic solvents such as methanol by Soxhlet extraction method. In the present study, a total of 23 different compounds were identified by GC-MS analysis using methanol extract, all the identified compounds were medicinally valuable for the treatment of various human ailments. In addition, all the phytochemical compounds were needed for further investigations on toxicological aspects for the development of new lead of therapeutic interest. Key words: Manilkara roxburghiana, Methanolic extract, Preliminary screening and GC-MS analysis. 1. Introduction  The plant kingdom represents an enormous reservoir of biologically active compounds with various chemical structures and these phytochemicals, often secondary metabolites present in smaller quantities in higher plants, include the alkaloids, steroids, flavonoids, terpenoids, tannins, and many others (Nonita et al., 2010). Modern medicine has evolved from folk medicine and traditional system only after through chemical and pharmaceutical screening (Boopathi and Sivakumar, 2011). Medicinal plants are the richest bio-resources of folk medicines and traditional systems of medicine; and food supplements, neutraceuticals, pharmaceutical industries and chemical entities for synthetic drugs (Ncube et al., 2008). The efficacy depends on the *Corresponding author: K. Saravanakumar E-mail: bavishmuruganatham1984@gmail.com Received: 02.04.2016; Revised: 18.04.2016; Accepted: 04.05.2016. use of proper plant part and its biological potency which in turn depends upon the presence of required quantity and nature of secondary metabolite in a raw drug (Vinoth et al., 2011; Savithramma et al., 2012). Herbal medicines have become more popular in the treatment of many diseases due to popular belief that green medicine is safe, easily available and with less side effects (Savithramma et al., 2011). Phytochemical studies have attracted the attention of plant scientists due to the development of new and sophisticated techniques. These techniques played a significant role in the search for additional resources of raw material for pharmaceutical industry (phytochemicals) (Mongole et al., 2010). Hence, GC-MS analysis was carried out in the present study with methanolic leaf extract of Manilkara roxburghiana to investigate the chemical constituents present in it. © 2015 Published by JPS Scientific Publications Ltd. All rights reserved Saravanakumar / Indo – Asian Journal of Multidisciplinary Research (IAJMR), 2(3): 615 – 619 616 2. Materials and methods Preliminary phytochemical screening Collection of plant materials and preparation of the extract The fresh leaves of Manilkara roxburghiana (Wight) Dubard were collected from the sacred grove of Silambur (Lat, 11.35 ºN; Long, 79.31ºE), Ariyalur District, Tamil Nadu, India. The specimen was botanically identified and confirmed by Rapinat Herbarium, St. Joseph’s College, Tiruchirappalli. The preserved plant specimens were submitted to the Department of Botany, Annamalai University, Annamalainagar, Tamilnadu for further reference. The leaves were chopped into small pieces, shade-dried and coarsely powdered by using a pulverizor. The powdered leaf was then subjected to successive extraction with organic solvents such as hexane chloroform and ethanol by Soxhlet method. The extracts were then collected and distilled off on a water bath at atmospheric pressure and the last trace of the solvents was removed in vacuo and stored at 4 ˚C. The extracts were dissolved in dimethylsulphoxide (5 % DMSO) to make the final concentrations at the time study. They were used for GC-MS analysis. A small portion of the dry leaf extracts were subjected to preliminary phytochemical screening to detect the presence of various phytoconstituents present in Manilkara roxburghiana (Trease and Evans, 1983; Harbourne, 1973). Gas Chromatography- Mass Spectrometry (GC-MS) analysis GC-MS analysis was performed with GCMS Clarus 500 Perkin Elmer Equipment. Compounds were separated on Elite-5 capillary column (Crossbond 5% Phenyl 95 % dimethylpolysiloxane). Oven temperature was programmed as follows: isothermal temperature at 60 ºC then increased to 200 ºC at the rate of 10 ºC/min, then increased up to 280 ºC at the rate of 5ºC/min. held for 9 min. Ionization of the sample components was performed in the Electron energy (70 eV). The helium was used as gas carrier (1ml/min), and 1.0 μl of sample was injected. The detector was Mass detector Turbomass gold Perkin Elmer. Table -1: Compounds identified in methanolic leaf extract of Manilkara roxburghiana S. No. 1. 2. 3. 4. 5. 6. Peak Name Name: 2H-Azepin-2-one, hexahydro-1-methylFormula: C7H13NO MW: 127 Name: 3,4-Furandiol, tetrahydro-, cisFormula: C4H8O3 MW: 104 Name: 2-Propyl-1-pentanol Formula: C8H18O MW: 130 Name: 2-Octanone Formula: C8H16O MW: 128 Name: 5,9-Dodecadien-2-one, 6,10-dimethyl-, (E,E))Formula: C14H24O MW: 208 Name: 1,2-Benzenediol Formula: C6H6O2 MW: 110 Retention Time (min) Peak Area % Peak area 4.63 28708 0.0992 6.34 137304 0.4744 6.55 1730096 5.9771 6.94 513893 1.7754 7.92 333359 1.1517 10.93 384297 1.3277 © 2015 Published by JPS Scientific Publications Ltd. All rights reserved Saravanakumar / Indo – Asian Journal of Multidisciplinary Research (IAJMR), 2(3): 615 – 619 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. Name: Hydroquinone Formula: C6H6O2 MW: 110 Name: 2-Methoxy-4-vinylphenol Formula: C9H10O2 MW: 150 Name: Phenol, 2,6-dimethoxyFormula: C8H10O3 MW: 154 Name: 1,2,3-Benzenetriol Formula: C6H6O3 MW: 126 Name: Propanedioic acid, propylFormula: C6H10O4 MW: 146 Name: D-Allose Formula: C6H12O6 MW: 180 Name: 1,3,5-Benzenetriol, dihydrate Formula: C6H6O3 MW: 126 Name: 4-((1E)-3-Hydroxy-1-propenyl)-2-methoxyphenol Formula: C10H12O3 MW: 180 Name: n-Decanoic acid Formula: C10H20O2 MW: 172 Name: Hexadecanal Formula: C16H32O MW: 240 Name: 3-t-Butyl-oct-6-en-1-ol Formula: C12H24O MW: 184 Name: 2,6,10-Trimethylundeca-1,3-diene Formula: C14H26 MW: 194 Name: Tridecanoic acid, methyl ester Formula: C14H28O2 MW: 228 Name: n-Hexadecanoic acid Formula: C16H32O2 MW: 256 Name: Phytol Formula: C20H40O MW: 296 Name: Squalene Formula: C30H50 MW: 410 617 12.88 673763 2.3277 13.11 179842 0.6213 13.96 42546 0.1470 15.08 11835200 40.8878 17.69 44548 0.1539 21.78 9668330 33.4018 22.54 904600 3.1252 24.18 49804 0.1721 24.52 291869 1.0083 26.32 316670 1.0940 26.96 153522 0.5304 27.41 94227 0.3255 28.56 132375 0.4573 29.67 420466 1.4526 33.05 156621 0.5411 47.78 853531 2.9487 © 2015 Published by JPS Scientific Publications Ltd. All rights reserved Saravanakumar / Indo – Asian Journal of Multidisciplinary Research (IAJMR), 2(3): 615 – 619 618 GC 15 02 X 0199C S3 27 02 15 % Scan EI+ TIC 9.04e7 15.08 126 100 6.56 57 29.67 73 21.78 60 6.94 58 2.82 44 5.38 117 22.54 126 10.93 12.88 7.93 110 110 43 47.78 69 33.05 34.86 37.18 39.45 41.67 43.83 45.94 73 73 71 73 73 73 73 26.32 43 28.56 74 0 Time 5.91 10.91 15.91 20.91 25.91 30.91 35.91 40.91 45.91 Fig – 1: GC-MS analysis of methanolic leaf extract of Manilkara roxburghiana The total running time for GC was 36 min and software Turbomass 5.2.0 was used in this GC-MS study. Identification of compounds All the compounds were identified from methanol extracts based on direct comparison of the retention times and their mass spectra with the spectra of known compounds stored in the spectral database, NIST (Version year 2005). 3. Result and Discussion The chemical constituents identified by the GC-MS analysis on various extracts of the leaves of Manilkara roxburghiana were enumerated along with Molecular Formula (MF), Molecular Weight (MW), Retention Time (RT), peak area, and peak area (%) was presented in Table - 1. In the methanolic leaf extract, totally 22 compounds were identified of which 2, 3-Benzenetriol (40.88 %), was found as major compound followed by other compounds namely, D-Allose (33.40 %), 2Propyl-1-pentanol (5.97 %), 1, 3, 5-Benzenetriol, dehydrate (3.12 %), Squalene (2.94 %) and Hydroquinone (2.32 %) respectively along with other minor constituents. 4. Conclusion In the present study, it has been concluded that the plant Manilkara roxburghiana is a potential source of biologically active compounds with pharmaceutical value. Further, the compounds identified were needed further study on the toxicological aspects including clinical trials to develop safe drug for the treatment of various human ailments. 5. References 1) Boopathi, A. C and R. Sivakumar. 2011. Phytochemical screening studies on the leaves and stem of Andrographis neesiana wight – An endemic medicinal plant from India. World Applied Science Journal, 12 (3): 307 - 311. 2) Mungole, A. J., R. Awati, A. Chaturvedi and P. Zanwar. 2010. Preliminary phytochemical screening of Ipomoea obscura (L) – A hepatoprotective medicinal plant. International Journal of Pharm Tech Research, 2: 2307 - 2312. 3) Ncube, N. S., A. J. Afolayan and A. I. Okoh. 2008. Assessment techniques of antimicrobial properties of natural compounds of plant origin: Current methods and future trends. © 2015 Published by JPS Scientific Publications Ltd. All rights reserved Saravanakumar / Indo – Asian Journal of Multidisciplinary Research (IAJMR), 2(3): 615 – 619 4) 5) 6) 7) 619 African Journal of Biotechnology, 7: 1797 1806. Nonita, P. P and Mylene, M. U. 2010. Antioxidant and cytotoxic activities and phytochemical screening of four Philippine medicinal plants. Journal of Medicinal Plant Research, 4: 407 - 414. Savithramma, N., M. Linga Rao and S. Ankanna. 2012. Preliminary phytochemical screening of some important medicinal plants. International Journal of Ayurvedic and Herbal Medicine, 2 (1): 139 - 145. Savithramma, N., M. Linga Rao and D. Suhrulatha. 2011. Screening of medicinal plants for secondary metabolites. Middle-East Journal of Science and Research, 8: 579 - 584. Vinoth, S., P. Rajesh Kanna, P. Gurusaravanan and N. Jayabalan. 2011. Evaluation of phytochemical, antimicrobial and GC-MS analysis of extracts of Indigofera trita. International Journal of Agricultural Research, 6 (4): 358 - 367. © 2015 Published by JPS Scientific Publications Ltd. All rights reserved