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The whole plant material of Corydalis adiantifolia Hook.f. & Thomson was investigated for biological activities e.g. antifungal, antibacterial, insecticidal, cytotoxic, anticancer and phytotoxic activities and preliminary phytochemical screening. The methanol extract and dichloromethane fraction of the plant species exhibited 5% inhibition each against the fungus Aspergillus flavus. The methanol extract and water and dichloromethane fractions exhibited non-significant antibacterial activity and they showed inhibition against Hela cell lines and insignificant insecticidal activity. To understand the bioactive profile of C. adiantifolia, phytochemical screening approach is considered effective. The samples including methanol extract and n-hexane, dichloromethane, ethyl acetate and water fractions were subjected to qualitative phytochemical screening for the presence of various phytochemicals i.e. alkaloids, flavonoids, saponins, diterprenes, triterpenoids, anthraquinones, anthranol glycosides, reducing sugars and phenols. The results exhibited the efficacy of methanol extract showing the presence of more phytochemicals in comparison to the fractions of C. adiantifolia. Moreover, as a result of phytochemical isolation, 8-acetonyldihydrosanguinarine (1), β-sitosterol (2) and β-sitosterol-β-D-glucoside (3) were isolated, purified, and characterized by spectroscopic methods. To the best of our knowledge, all this study was carried out for the first time on C. adiantifolia.