GC-MS ANALYSIS OF METHANOLIC EXTRACT OF LEAVES OF RHODODENDRON CAMPANULATUM Short Communication SAKSHI PAINULI, NISHANT RAI, NAVIN KUMAR* Department of Biotechnology, Graphic Era University, Clement Town, Dehradun 248001, Uttarakhand, India Email: navinkumar.bt@geu.ac.in Received: 03 Sep 2015 Revised and Accepted: 02 Nov 2015 ABSTRACT Objective: Rhododendron campanulatum is a native of high altitude and is known for its medicinal properties. The present study is aimed to identify the phytochemical constituents in the leaf extract of R. campanulatum using Gas Chromatography and Mass Spectroscopy (GC-MS). Methods: The methanolic leaf extract was prepared using Accelerated Solvent Extraction system at room temperature and high pressure. Phytochemical screening of methanolic extract of R. campanulatum was performed using GCMS-QP2010 Plus (Shimadzu, Kyoto, Japan) and the spectrum was interpreted on the basis of the databases of National Institute Standard and Technology (NIST11LIB) and WILEY8LIB. Results: The GC-MS analysis revealed the presence of 49 phytochemical compounds in the methanolic leaf extract. Baccharis oxide (9.99%), betuligenol (8%), alpha and beta-amyrin (7.38 and 2.64%), geranyl acetate (5.91%), (R)-(-)-14-methyl-8-hexadecyn-1-ol (5.19%) and phthalic acid (5.16%) were identified as major constituents. Conclusion: The methanolic leaf extract of R. campanulatum contains various phyto-compounds of pharmaceutical and industrial importance. Keywords: Rhododendron campanulatum, Gas Chromatography-Mass Spectrometry, Phytochemical compounds. Rhododendron genus is comprised of about 1025 species, which are mostly found at higher altitudes [1, 2]. In India, there are around 80 species and 14 subspecies of Rhododendron, distributed in the Himalayan region at the altitude ranging 1500-5500 meters [3]. Different species of Rhododendron are known for their ethnopharmacological values [4]. R. campanulatum is a very important member of the genus Rhododendron, which is known for its traditional medicinal importance for the different ailments like body ache, sore throat, digestion, skin diseases, rheumatism, syphilis, cold and fever, etc. [4-9]. R. campanulatum is found at altitudes between 2500-4300 m [2]. There are very few scientific studies on phytochemical constituents and therapeutic potential of the R. campanulatum. Analysis through High Performance Thin Layer Chromatography (HPTLC), High-Performance Liquid Chromatography (HPLC) and Nuclear Magnetic Resonance (NMR) have shown the presence of epicatechin, syringic acid, quercetin, chlorogenic acid, gallic acid, proto catechic acid and oleanane triterpenoid in the different extracts of leaves of R. campanulatum [6, 9, 10]. Antibacterial properties have also been shown in the leaf extract of R. campanulatum [8]. In the present study, we attempted to identify the phytochemical constituents in the methanolic leaf extract of R. campanulatum using GC-MS method of analysis. R. campanulatum leaves were collected from the Govind ghat region of Chamoli District, Uttarakhand, India. Plant sample was authenticated by Department of Botany at H. N. B Garhwal University (Srinagar), Uttarakhand. The Herbarium specimen was deposited in the Department of Botany; H. N. B Garhwal University vide voucher number GUH-0743. The leaves of R. campanulatum were shade dried at sterilized condition and pulverized with the grinder. The powder was successively extracted with methanol (100%) in the Accelerated Solvent Extraction System, which was equipped with a solvent controller unit (ASE350, Dionex Corporation, Sunnyvale, CA, USA) and further the extract was subjected to lyophilization [11]. The lyophilized leaf extract was then stored at 4 °C for further investigation. 1 µl of prepared sample (1 mg/ml lyophilized extract in methanol) was applied for GC-MS analysis. GC-MS analysis was performed at University Science Instru- mentation Centre, AIRF, Jawaharlal Nehru University, Delhi. For GC- MS analyses of plant extract, GC-MS QP2010 Plus (Shimadzu, Kyoto, Japan) system was utilized. The system was equipped with an auto injector (AOC-20i), head space sampler (AOC-20s), a mass selective detector with an ion source (220 °C) and an interface (260 °C). Rtx-5 MS capillary column having 30 mm X 0.25 mm of length X diameter and 0.25 µm of film thickness was used for MS analyses. The mass range of 40-650 m/z with 1,000 ev of the threshold was purposed. The injector was set in the split injection mode having 250 °C of temperature. The ratio applied for split mode was 10.0. The starting temperature was adjusted to 80 °C (3 min), which afterwards increased to 280 °C with a ramp rate of 10 °C/min. Helium (>99.99 %) with 40.5 cm/s of linear velocity was employed as a carrier gas. The system was programmed with 16.3 ml/min of total flow rate and 1.21 ml/min of column flow. Fig. 1: GC-MS chromatogram of the constituents of methanolic leaf extract of R. campanulatum International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 7, Issue 12, 2015 Innovare Academic Sciences