World Journal of Pharmaceutical Research Karpe et al. World Journal of Pharmaceutical Research SJIF Impact Factor 7.523 Volume 6, Issue 8, 1164-1172. Research Article ISSN 2277–7105 GC-MS ANALYSIS OF CRUDE EXTRACTS OF MOULLAVA SPICATA (DALZ.) NICOLSON Dnyaneshwar G. Karpe* and Shamrao P. Lawande P.G.Department of Chemistry, Shri Chhatrapati Shivaji College Shrigonda, Dist-Ahmednagar (MS) - 413701. Article Received on 25 May 2017, ABSTRACT M.spicata is candy corn plant, it shows various biological activities. Revised on 14 June 2017, Accepted on 05 July 2017 The investigation was carried out to determine the chemical DOI: 10.20959/wjpr20178-8945 constituents of M.spicata aerial parts using RTx-5MS Gas – chromatography – Mass spectroscopy, while the mass spectra of the *Corresponding Author compounds found in the extract was matched with the National Dnyaneshwar G. Karpe Institute of Standards and Technology (NIST) library. GC-MS analysis P.G.Department of of petroleum ether (MSPE), ethyl acetate (MSEA) and methanol Chemistry, Shri (MSME) extracts of aerial parts of M.spicata revealed the existence of Chhatrapati Shivaji College Shrigonda, Dist- various chemical constituents. The results of this study offer a platform Ahmednagar (MS) - of using M.spicata aerial part as a herbal alternative for various 413701. diseases. KEYWORDS: GC –MS analysis, Moullava spicata, Chemical constituents. INTRODUCTION In many industrialized countries herbal medicines are gaining popularity as alternative and complimentary therapies. Some of the plants are used as food or medicine. These plants exhibit a wide range of biological and pharmacological activities such as antibacterial, antifungal, antimalarial and antituberculosis. The secondary metabolites of plants provides humans with numerous biologically active products which has been used extensively as drugs, foods, additives, flavors, insecticides, colorants, fragrances and chemicals. Plants are a rich source of secondary metabolites with interesting biological activities. In general these secondary metabolites are an important source with a variety of structural arrangements and properties. Essential oils are known as to possess remarkable biological properties.[1] In www.wjpr.net Vol 6, Issue 8, 2017. 1164 Karpe et al. World Journal of Pharmaceutical Research developing countries, herbal medicines are gaining popularity as alternative and complimentary therapies. The plants exhibit a wide range of biological and pharmacological activities.[13] Plants are rich source of secondary metabolites with interesting biological activities.[13] The herbal medicines occupy distinct position right from the primitive period to present day.[15] India has wealth of medicinal plants, most of which have been traditionally used in Ayurveda.[15] Phytochemical constituents are responsible for medicinal activity of plant species.[16] Medicinal plants are the richest bio- resources of folk medicines and traditional systems of medicine and food supplements, nutraceuiticals, pharmaceutical industries and chemical entities for synthetic drugs.[22] Modern medicine has evolved from folk medicine and traditional system only after through chemical and pharmacological screening. India is the birth place of renewed system of indigenous medicine such as Siddha, Ayurveda and Unani. The synthetic drugs have many side effects than often lead to serious complications.[17] Comparing to modern medicine the herbal medicine was the lifesaving drug. Among 4, 00,000 plant species only 6% of the plants are studied for their biological activity and only few have been phytochemically investigated. It shows that the investigation is needed for many medicinal plants for its activity and pharmacological properties.[17] World Health Organization (WHO) has estimated that nearly 80% of the total population in the developing countries relies on medicinal plants for health care. M.spicata is used to cure many diseases; it shows prominent antioxidant activity.[9,11] In present investigation M.spicata aerial part extracted in different solvents with their increasing polarity. GC – MS analysis of crude extracts was done. MATERIALS AND METHODS Collection of Plant Material The fully matured M.spicata aerial parts were collected in Feb.2015 from Radhanagari, Kolhapur (MS). Plant material was identified and authenticated by Mrs.Dr.A.S.Upadhye, Scientsit, Agharkar Research Institute, Pune (MS). Extraction of plant material Collected plant material was cleaned; shade dried for 10 days, powdered in pestle and mortar .500 gm powdered plant material is extracted in petroleum ether for six hours in soxhlet extractor then filtered through whatmann filter paper No.41 along with 2 gm sodium sulfate to remove the sediment and traces of water in the filtrate. The filtrate is then concentrated on rotary evaporator (Buchi. Rotavapor, R-3) at reduced pressure. Thimble was dried in an oven www.wjpr.net Vol 6, Issue 8, 2017. 1165 Karpe et al. World Journal of Pharmaceutical Research at 600C and then same plant material extracted in ethyl acetate using soxhlet extractor, solvent is evaporated, thimble was dried at 750C in an oven. The same plant material was further extracted in methanol by soxhlet apparatus, dry extracts are labeled as MSPE, MSEA, and MSME. GC-MS analysis GC MS analysis was carried out on Shimadzu – GCMS QP-2010 Ultra model at S.P.Pune University, Pune. This system comprising a gas chromatograph interfaced to a mass spectrometer instrument employing the following conditions. Column RTx-5MS fused silica column (30 m X 0.25 mm X 0.25 μm composed of 5% Phenyl / 95% dimethylpolysiloxane) operating in electron impact mode at 70 ev, helium (99.999%) was used as carrier gas at a constant flow of 1 ml / min and an injection volume of 0.5 μl was employed (split ratio 25:1) Injection temperature 2400C ion source temperature 2000C. The oven temperature was programmed from 750C (isothermal for 3 min) with an increase of 100 C / min to 2400C, ending with 9 min , isothermal at 2900C , Mass spectra were taken at 70 ev, a scan interval of 0.5 seconds and fragments from 40 to 440 Da. Total GC running time is 26.83 min. RESULTS AND DISCUSSION Gas chromatography mass spectroscopy analysis of crude extracts of M.spicata was done. The total ion chromatogram (TIC) of petroleum ether, ethyl acetate and methanol extract of M.spicata showing the GC-MS profile of the compounds identified is given in the figures 1, 2,3 respectively. The peaks in the chromatogram were integrated and were compared with the database of spectrum of known components stored in the GC-MS library. The detailed GCMS analyses of the extracts are given in Table 1, Table 2, Table 3 respectively. Figure 1: Total Ion Chromatogram (TIC) of petroleum ether (MSPE) extract of M.spicata. www.wjpr.net Vol 6, Issue 8, 2017. 1166 Karpe et al. World Journal of Pharmaceutical Research Figure 2: Total Ion Chromatogram (TIC) of Ethyl acetate (MSEA) extract of M.spicata. Figure 3: Total Ion Chromatogram (TIC) of Methanol (MSME) extract of M.spicata. www.wjpr.net Vol 6, Issue 8, 2017. 1167 Karpe et al. World Journal of Pharmaceutical Research Table 1: Phytochemicals identified in the petroleum ether extract of M.spicata. Ret. Time (min.) Name of the Compound M.F M.W. Area % Compound Structure OH 7.799 2,4 –ditertiary butyl phenol 8.150 Dodecanoic Acid 8.370 1-Heptadecene 10.020 10.305 13.020 Tetradecanoic Acid 1-Nonadecane (+) Ascorbic Acid 2,6dihexadecanoate C14 H22O 206 4.71 C12 H24O2 238 2.65 C17 H34 238 5.97 C14 H28O2 228 12.80 C19 H38 266 2.83 C38 H68O8 652 13.31 O OH H3C O HO O O H HO O OH O O O 13.500 (Z)9-Tricosene 17.810 Oleic Acid C23 H46 322 2.89 C18 H34O2 282 5.12 HO O O 20.170 Gingerol C17 H26O4 294 OH 3.96 HO OCH 3 23.300 6-(3,5-Dimethyl furan -2yl)-6methyl-hept-3ene-2-one C14 H20O2 220 2.35 O O www.wjpr.net Vol 6, Issue 8, 2017. 1168 Karpe et al. World Journal of Pharmaceutical Research Table 2: Phytochemicals identified in the ethyl acetate extract of M.spicata. Ret.Time Name of the (min.) Compound M.F M.W. Area % Compound Structure O 6.445 3-cyclohexene1-methanol,α,α4-trimethyl acetate 6.700 1-pentadecene O C12 H20O2 196 1.66 C15 H30 210 0.87 OH 7.800 2,4-dit.butyl phenol 8.365 1-Heptadecene 12.905 (+)-Ascorbic Acid 2,6dihexadecanoate C14 H22O 206 0.61 C17 H34 238 1.52 652 1.91 C38 H68O8 O O H HO O OH O O O 22.015 22.505 3-(6-methyl-3Pyridyl)-1phenyl-5(-ptolyl)-2pyrazoline C22 H21N3 Friedelan-3-one C30 H50O 327 59.52 N N N 426 100 O www.wjpr.net Vol 6, Issue 8, 2017. 1169 Karpe et al. World Journal of Pharmaceutical Research Table 3: Phytochemicals identified in the methanol extract of M.spicata. Ret. Time (min.) Name of the Compound M.F M.W. Area % Compound Structure OH 7.795 2,4-ditert.butyl phenol C14H22O 206 9.74 O 12.240 22.090 Hexadecanoic Acid methyl ester Diisooctyl phthalate C17H34O2 C24H38O4 270 4.48 H3C 390 10.26 (CH2)13 O OCH3 OO O 25.105 3,5-Dichloro4(dodecylsulfanyl)C19H31O2NS 2,6-dimethyl pyridine S 375 6.96 Cl Cl N The GC MS analysis of crude extracts of M.spicata was performed using Shimadzu – GCMS QP-2010 Ultra. MSPE shows presence of ten compounds, MSEA shows presence of eight compounds and MSME shows presence of four compounds. Some of the peaks are unidentified. ACKNOWLEDGEMENT Authors are thankful to Principal, Shri Chhatrapati Shivaji College, Shrigonda, DistAhmedngar (MS) for providing necessary facilities to carry out this research work. Authors are also thankful to Director, Central Instrumentation Facility (CIF), Savitribai Phule Pune University, Pune (MS) for carrying out GC-MS analysis of samples. REFERENCES 1. Waghole R. J.and Naik D. G.“Chemical composition and Antifungal properties of essential oil from Tetrastigma sulcatum (Law.) Gamble leaves. Journal of Essential oil bearing plants Taylor and Francis, 2016; 19(3): 568-573. www.wjpr.net Vol 6, Issue 8, 2017. 1170 Karpe et al. World Journal of Pharmaceutical Research 2. Cooks, The flora of Bombay Presidency, Taylor and Francis, London, 1903; 1: 416. 3. Kirtikar, K.R., Basu, B.D., Indian medicinal plants, Lalit Mohan Basu, Allahabad, 1933; 2: 853. 4. Chopra, R.N., Nayar, S.L., Chopra, I.C., Glossary of Indian Medicinal Plants, Council of Scientific and Industrial research, New Delhi, 1956; 257. 5. Nadkarni, K.M., Indian Materia Medica, Vol.I, Bombay Popular Prakashan, 1976, 1290. 6. The useful plants of India, Anonymous, Council of Scientific and Industrial Research, New Delhi, 1992; 686. 7. Joshi, D.V., Tamhane, R.G., Datta, N.K., Current Science, 1957; 26(25): 147-148. 8. Shamak, G., Shenoy, R.P., Manjunatha, S.M., Vinayak, K.S., Food Chemistry, 2009; 115(2): 631-634. 9. Lohith, K., Vijay, R., Pushpalatha, K.C., Joshi, C.G., Phytochemical and Antioxidant evaluation of moullava spicata (Dalzell) Nicolson leaf extract, Annual Research and Review in Biology, 2014; 4(1): 188-197. 10. Sadasivam,S., Manikam, A., Biochemical Methods,3rdedition,New Age International Publishers, New Delhi, 2008; 203-211. 11. Lohith, K., Vijay, R., Joshi K.C., Chandrasekhar G., In vitro cytotoxic study of Moullava spicata (Dalz.) Nicolson leaf extract, Indian journal of forensic medicine and toxicology, July-Dec 2013; 7(2): 182. 12. Mooza Al-Owaisi, Nora Al Hadiwi, Shah Alam Khan, “GC –MS analysis , determination of total phenolics, falconoid content and free radical scavenging activities of various crude extracts of moringa peregrine (Forssk) fiori leaves”. Asian Pacific Journal of Tropical Biomedicine, 2014; 4(12): 964-970. 13. R. Meenatchi Ammal, G.Viji Stella Bai, “GC-MS Determination of Bioactive Constituents of Heliotropium indicum Leaf”, Journal of Medicinal plants studies, 2013; 1(6): 30-33. 14. Elizabeth Thomas, Aneesh T.P, Della Grace Thomas, R.Anandan, “GC-MS analysis of phytochemical compounds present in the rhizomes of Nervilia aragoana Gaud”, Asian journal of Pharmaceuitical and Clinical Research”, 2013; 6(3): 68-74. 15. G.M. Krishnaiah, Prashant G.K, “Phytochemical studies and GC-MS analysis of the Leaf Extracts of Melia Azedarach Linn”, International Journal of Advancement in Engineering Technology, Management and Applied Science”, 2014; 1(6): 48-54. www.wjpr.net Vol 6, Issue 8, 2017. 1171 Karpe et al. World Journal of Pharmaceutical Research 16. K.Kalimuthu, R.Prabakaran, “Preliminary Phytochemical screening and GC-MS analysis of methanol extract of Ceropegia pusilla, “International Journal of Research in Applied, Natural and Social Sciences, 2013; 1(3): 49-58. 17. Rukshana MS, Doss A , Kumari Pushpa Rani TP , “Phytochemical Screening and GCMS Analysis of Leaf extract of Pergularia daemia (forssk) Chiov”, Asian Journal of Plant Science and Research, 2017; 7(1): 9-15. 18. S. Gopalkrishnan, E.Vadivel, “GC-MS analysis of some bioactive constituents of Mussaenda frondosa Linn”, International Journal of Pharma and Biosciences, 2011; 2(1): 313-319. 19. Raja Rajeshwari. N, Rama Lakshmi. S, Muthuchelian. K, “GC-MS analysis of bioactive components from the ethanolic leaf extract of Canthium dicoccum (Gaertn) Teijsm and Binn.” Journal of Chemical and Pharmaceuitical Research, 2011; 3(3): 792-798. 20. Arun Thangavel, Senthilkumar Balkrishnan, Aarthy Arumugam , Senbagam Duraisamy, Sureshkumar Muthusamy, “African Journal of Pharmacy and Pharmacology, 2014; 8(5): 126-135. 21. Enas J. Kadhim , Duha A, Al-Shammaa,”Phytochemical Characterization using GC-MS Analysis of methanolic extract of Moringa oleifera (family Moringaceae) Plant cultivated in Iraq”, Chemistry and Materials Research, 2014; 6(5): 9-27. 22. Ncube NS, Afolayan AJ and Okoh AI, “Assessment techniques of antimicrobial properties of natural compounds of plant origin: Current methods and future trends.”African J. of Biotechnol, 2008; 7: 1797-1806. www.wjpr.net Vol 6, Issue 8, 2017. 1172