Inhibitory potential of the dichloromethane, ethyl acetate, ethanol, and aqueous extracts of Viola odorata L. (VO) was investigated against tyrosinase (TYR) and cholinesterases by microplate assays. The antioxidant activity was tested using six in vitro assays. Only the ethanol extract inhibited TYR (80.23 +/- 0.87% at 100 mu gmL(-1)), whereas none of them were able to inhibit cholinesterases. The extracts were more able to scavenge NO radical (31.98 +/- 0.53-56.68 +/- 1.10%) than other radicals tested, and displayed low to moderate activity in the rest of the assays. HPLC analysis revealed that the aqueous extract of VO contained a substantial amount of vitexin (18.81 +/- 0.047mgg(-1) extract), while the ethanol extract also possessed rutin (1.31 +/- 0.013mgg(-1) extract) and vitexin (4.65 +/- 0.103mgg(-1) extract). Furthermore, three flavonoids (rutin, isovitexin, and kaempferol-6-glucoside) were isolated from the ethanol extract. This is the first report on TYR inhibitory activity of VO as well as presence of vitexin and isovitexin in this species. Copyright (c) 2015 John Wiley & Sons, Ltd.