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From the Preliminary Phytochemical screening we inferred the presence of Pentacyclic triterpenoids, Alkaloids, Carbohydrates, Saponins, Phytosterols, Tannins and gums as Major compound in this whole plant material. The pet ether extract was found to consist of -amyrin -amyrin acetate sitosterol, sitosterol acetate. These compounds are identified by GC-MS and NMR studies. The crude extracts and column-separated samples are subjected to anti-microbial activity test and results provided the anti-microbial activity of the whole plant. The crude whole plant material subjected to the antioxidant properties and its shows positive results for the anti-oxidant properties. So the plant has it own antioxidant potential. Hence from the present investigation it is evident that the whole plant material (Cleome gynandra) has a high content of all types of vitamins, biochemical factors such as carbohydrates, proteins, fats and also the various antioxidative enzymes such as catalase, superoxide dismutase and glutathione.