In this study, we have aimed to analyze the phytochemical composition of this plant and the concentration of strychnine and brucine. The identification of bioactive compounds was done by GC-MS with NIST Library. Strychnine and Brucine were quantified using HPLC. Twenty one medicinal bio active compounds were identified from the Strychnos nux-vomica leaf ethanolic extract. Strychnine is showing 28.43% purity and brucine was not detected in GCMS analysis. Quantified the concentration of strychnine (0.6 mg in 500mg of extract) and brucine (1.6 mg in 500mg of extract) was done by HPLC against Strychnine and Brucine standard. These compounds are having natural properties of Anti-inflammatory, Hypocholesterole, Cancer preventive, Hepatoprotective, Antimicrobial, Antioxidant, Cardio protective, Antiaging, Antialzheimeran, Antidermatitic, Immunostimulant, Anthepatotoxic, biosynthesis of steroid hormones, Nematicide, Antiandrogenic, 5-alpha reductase inhibitor, antipsychotic, analgesic, apoptotic effect, antidepressant, antidote for snake poisoning and diabetic activity.