Rauvolfia serpentina, commonly known as Sarpgandha, is a plant well-known for being utilized for several medicinal purposes. Unrestricted collection from natural stands and overexploitation for medicinal and traditional purposes have rendered it endangered, hence there is necessary requirement for the development of cultivation protocols for mass propagation and sustained utilization of the plant. In the present study, in vitro culture of the apical parts of R. serpentina on MS (Murashige and Skoog) medium enriched with NAA resulted in development of callus, whereas multiple shoot regeneration along with callus development was achieved on medium combination MS + NAA + Kn and MS + NAA + BAP. MS + 4µM NAA + 4µM BAP was found to be most optimum media composition for regeneration of shoots and callus. Among different media combinations utilized for in vitro rooting, a maximum of 82.6 % explants developed in vitro roots on ½ MS + 12µM NAA. Gas chromatography-mass spectrometry (GC-MS) analysis of methanolic extract prepared from leaves of wild and micropropagated plants of R. serpentina revealed the presence of 38 and 48 phytocompounds, respectively. 9, 12-Octadecadienoic acid, Methyl linoleate, Methyl stearate, Hexadecanoic acid, methyl ester, Linoleic acid, Ergost-4, 7, and 22-trien-3.alpha.-ol were some of the major compounds found to be present in the leaves of wild plants, and Linoleic acid, methyl ester, Cis-Linoleic Acid Methyl Ester, Methyl elaidate, hexadecanoic acid, and methyl ester were major compounds found to be present in in vitro raised plants. Many of the compounds detected have been known to possess 1 or more biological or pharmacological activities.