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Abstract Botanical study including macro- and micro-morphological studies of different organs of Melilotus indicus (L.) All. were achieved for authentication and identification of the plant in the entire and powdered forms. The phytochemical study was established including preliminary screening, investigation of the essential oil, quantitative determination of the active constituents, HPLC fingerprint analysis of different plant extractives with assignment of the isolated compounds, UPLC-Mass profile of secondary metabolites of 80% methanolic extract and investigation of the polysaccharide content.Seven compounds have been isolated and identified via physical, chromatographic and spectral data namely; a hydrocarbon (nonacosane), a fatty acid (palmitic acid), a benzopyrane (coumarin), a flavonoid (kaempferol), flavonoid glycosides (trifolin and robinin) and o-coumaric acid.UPLC/Ms analysis led to the identification of 14 compounds from which 5 compounds were identified for the first timein genus Melilotus. Defatted 80% methanol extract and its subfractions have weak antioxidant activity. Methylene chloride subfraction showed cytotoxic activity on liver and colon human cell lines and ethylacetate subfraction showed a strong anticoagulant activity |