Background: Plant extracts contains multiple active constituents which leads to the production of new drugs from plants and chemicals derived from various parts of plants. The objective of present study was to investigate the GC-MS analysis and antibacterial activity of L. inermis leaves extracts. Material and Methods: Crude methanol extract and its fractions were tested for the presence of active phytochemicals and GC-MS analysis of hexane; ethyl acetate and aqueous methanol fractions was performed. Antimicrobial activity against six bacterial strain’s Escherichia coli, Staphylococcus aureus, Bacillus subtelis, Salmonella typhi, Klebsiella and Pseudomonas aeruginosa was also tested. Results: Phytochemical screening of extract confirmed the presence of carbohydrates, glycosides, quinones, steroids and phenol. In GC-MS chromatograms, 56, 108 and 19 peaks were obtained and out of these, 13, 17 and 7 compounds were identified in hexane, ethyl acetate and aqueous methanol fractions, respectively. Conclusion: For best of our knowledge in L. inermis leaves extract, Celidoniol and Vitamin E has not been reported earlier in hexane fractions. While 2, 3 dihydrobenzo furan, 1-H indole -1, 3 (2H)-dione, 1 (3H)-Isobenzofuranone, 1H Isoindole-1, 3 (2H) Dione, Napthelene, 2 ethoxy, 2 (4H) Benzofuranone, Vitamin E, Benzene, 1 isocyano 4 methyl are also identified for the first time in ethyl acetate fraction. Also, in aq. Methanol fraction 1(3H)-isobenzofuranone, Squalene and Vit E were not previously identified. Highest antibacterial activity was confirmed in crude methanol extract which might be due to all the antibacterial compounds present in its fractions. The present study helped in identifying phytoconstituents present in the extract and its fractions which are responsible for various biological and antibacterial activities.