This study was conducted to evaluate the depigmenting properties of ethanol extract from a Fritillaria verticillata Willd. (EFV) in B16F10 cells. Fritillaria verticillata Willd., a perennial herbaceous plant, has been used as a stimulator of mammary gland, expectorant, blood pressure depressant, antitussive agents in Korean herbal medicine. In the present study, we observed that melanin synthesis of B16F10 cells were significantly decreased by EFV without cytotoxicity. However, EFV could not suppress tyrosinase activity in B16F10 cells and mushroom tyrosinase activity. Furthermore, EFV did not effect the protein expression of tyrosinase, tyrosinase-related protein -1 (TRP-1), and TRP-2. These results suggest that EFV inhibited melanin synthesis and the hypopigmentary effect of EVF was not due to regulation of tyrosinase protein.