Semecarpus anacardium (SA) is a high medicinal value plants. The tribal area people has been using nuts of SA for the treating tuberculosis since long time but active chemical constituent are still unknown. Present study was carried out to isolate, identify and evaluate bioactive compounds of SA nuts extracted using Agilent 7890A GC-MS. Solvent extraction of SA nuts was done with petroleum ether, ethyl acetate, methanol and finally with water. All the extracts were tested for their bioactivity against potential pathogen Mycobacterium tuberculosis in collaboration with Central Drug Research Institute, Lucknow, India. The extract which was found to be highly activity in terms of MIC against M. tuberculosis, was subjected for the Gas chromatography and mass spectroscopy. Also potential extract was subjected to phytochemical analysis. Proximate analysis and nutritive value determination was done in SA nuts powder. Also potential extract was subjected to column chromatography over silica gel for fractionations. After sequential elution with distilled water, methanol and their combinations, two fractionations were collected and subjected to TLC for checking proper separation before analyzing by GC-MS. Water extract showed potential with MIC 6.25 μg/ml against M. tuberculosis during in vitro bioassay experiment which is the pointing out towards potential of this plant against this microorganism. Phytochemical analysis results showed the presence of Alkaloids, Carbohydrate, Lipids, Saponins, Flavonoids, Tannins, Sterols and Triterpenes. Total nutritive value in 100g nuts was found 359.2 kilo caloric. In chromatogram of fraction 1 (SA/WE-01), out of six peaks, four peaks were analyzed and identified as per the fragmentation pattern as Benzoic acid 4-ethoxy ethyl ester, 2- lacosanol , n-hexadecanoic acid and n-Heneicosane. In fraction 2 (SA/WE-02) only one peak analyzed and identified as per the fragmentation pattern as Phenol, 2,4-bis (1,1-dimethylethyl). Analysis and identification of presence of compound in these extracts was done using database of NIST library.