GC-MS Analysis of n-hexane Extract of Stem Bark of Symplocos

Context: The stem barks of the plant Symplocos crataegoides Buch.-Ham. ex D. Don (syn. Symplocos paniculata (Thunb.) Miq.), Fam. Symplocaceae is extensively used in Indian medicine under the names of Lodhra/Patikalodhra. Mainly it is used to cure uterine complaints, vaginal and menstrual disorders. Aim: To investigate the phytochemicals from the n-hexane extract of the stem bark of Symplocos crataegoides Buch.-Ham. ex D. Don., using GC-MS analysis. Materials and Methods: Stem bark of S.crataegoides was extracted by Soxhlet extraction method using n-hexane. The extract was injected by splitless injection mode into the GC MS 5975 C Agilent equipped with a QP5000 (quadrupole) Gas Chromatography Mass Spectrometer. Results: Identification of 57 compounds from n-hexane extract. Those compounds were identified by close matches with standard MS spectra and compared with NIST 11 and WILEY library data. Undecane (7.51%) was found as major compound followed by Isopropyl myristate, Dodecane, 1,2,4-trimethyl-benzene, Octacosane, 2-methyl-decane, 2-ethyl-1,2-dimethyl-benzene, 1,2,3,5tetramethyl-benzene etc., other constituents were found to be in traceable quantities. Conclusion: GC-MS analysis of S. crataegoides revealed certain interesting facts of presentation of various phytoconstituents in the stem bark. The presence of various phytoconstituents contributes to the medicinal activity of the plant.

crataegoides as ingredients and Nyagrodhadi curna contain only Symplocos racemosa. 9ost of the Books on Indian Materia Medica equate Symplocos racemosa as well as S. crataegoides as the botanical source of Lodhra or Rodhra. 2 Two varieties of Lodhra are described in the texts viz.Sabara lodhra and Patika lodhra.Sabara lodhra is equated with Symplocos racemosa and Patika lodhra is equated with Symplocos crataegoides. 8otanically S. racemosa and S. crataegoides are different species called by similar vernacular name Lodhra.The petroleum ether (60-80⁰) extract of leaves of Symplocos crataegoides shows the presence of Octacos-1-ene, Stigmasterol and Lupeol, whereas ethanolic extract yields Salirepin. 10The following compounds isolated from the ethanolic extract of the stem bark of Symplocos crataegoides (syn: S. paniculata) were reported.4-(8-hydroxyethyl) cyclohexan-1-oic acid; androst-5( 6 11 The present investigation is carried out to determine the possible chemical components of n-hexane extract of stem bark of S. crataegoides by GC-MS analysis.

Procurement of Plant Materials
The authentic samples of stem bark of Symplocos crataegoides Buch.-Ham.ex D. Don.(Figure 1) was supplied by Dr.G.C.Joshi, Research Officer, Regional Research Institute of Himalayan Flora, CCRAS, Thapala, Ranikhet -263645, Almora dist., Uttarakhand.The museum specimen (C/222B SB26) is deposited in the department of Pharmacognosy, Captain Srinivasa Murthy Regional Ayurveda Drug Development Institute, Arumbakkam, Chennai, India for future reference.

Preparation of Extract
The shade dried and coarsely powdered (10/44) stem bark of S. crataegoides (4 g) were extracted using n-hexane (AR) by soxhlet extraction method.The extract was filtered through Whatman No.1 filter paper and concentrated in vacuum to constant weight.The yield of the extract is 0.4 %.The n-hexane extract of the plant was used for GC-MS analysis.

Gas Chromatography -Mass Spectrometer Conditions
Gas chromatography combined with mass spectroscopy is a preferable methodology for routine analysis of compounds.n-hexane extract was injected by split less injection mode into the GC MS 5975 C Agilent equipped with a QP-5000 (quadrupole) mass spectrometer, fitted with a DB -5 MS ultra inert 30 m x 250 μm, film thickness 0.25 μm capillary GC column, coated with polydimethyl siloxane.Helium was the carrier gas at a flow rate of 1.5 ml/min.The injector port temperature was 250°C, the detector temperature was 250°C and the oven temperature was maintained at 70°C for 3 mins, 10°C/min up to 300°C for 9 mins.The ionization voltage was 70 eV.It is separated into various constituents with different retention time which are detected by mass spectrophotometer.The chromatogram shows a plot of intensity against retention time was recorded by the software attached to it.The constituents were identified by comparing retention times of the GC peaks with those of reference compounds run under identical conditions and fragmentation patterns in mass spectra were matched with those of the NIST -11 & WILEY library and published mass spectra. 12

RESULTS AND DISCUSSION
GC-MS analysis of the n-hexane extracts of the stem bark of S. crataegoides is given in Table 1.Fifty seven compounds were identified by comparison with the authentic spectra obtained from GC-MS library (NIST -11 and WILEY).Even though several peaks were present in the GC-MS chromatogram (Figure 2), the identification of only 57 constituents was established by comparison with the authentic spectra obtained from GC-MS library with the SI factor.Qualitative GC-MS analysis of n-hexane extract results revealed that Undecane (7.51 %, Figure 3 & 4) was found as major compound.It is a liquid alkane hydrocarbon with the chemical formula CH 3 (CH2) 9 CH 3 .It is reported as a mild sex attractant for various types of moths and cockroaches, and an alert signal for a variety of ants. 13It has 159 isomers.Isopropyl myristate is the ester of isopropanol and myristic acid was found second major compound in the extract of the plant (7.45%, Figure 5 & 6).It is used in cosmetic and topical medicinal preparations where good absorption through the skin is desired.It is also used as a pesticide-free treatment against head lice which works by dissolving the wax that covers the exoskeleton of head lice, killing them by dehydration.Dodecane is present third major percentage present in the extract (4.60%, Figure 7         be in small quantities.All the above 57 compounds were identified and reported for the first time from the stem bark of the plant.

CONCLUSION
In the present study fifty seven chemical constituents have been identified from n-hexane extract of the stem bark of S. crataegoides by Gas Chromatogram Mass Spectrometry (GC-MS) analysis.The presence of various phytochemicals contributes to the medicinal activity of the plant.

Table 1 :
& 8).It is a liquid alkane hydrocarbon with the chemical formula CH 3 (CH 2 ) 10 CH 3 , an oily liquid of the paraffin series.It has 355 isomers.Contd.