In order to develop a new skin whitening agent, safflower (Carthamus tinctorius L.) seeds were evaluated for melanogenesis inhibitory activity and its active principles were identified following activity-guided isolation. The 80% aqueous methanol extract and ethyl acetate fraction from safflower seeds showed a significant inhibition for mushroom tyrosinase. Three active compounds, N-feruloylserotonin, N-(p-coumaroyl)serotonin, and acacetin, were isolated from the ethyl acetate fraction as the active principles. Compared with arbutin (IC₅₀=0.223 mM), the IC₅₀ values of these compounds were 0.023, 0.074, and 0.779 mM for N-feruloylserotonin, N-(p-coumaroyl)serotonin, and acacetin, respectively. It was also found that N-feruloylserotonin and N-(p-coumaroyl)serotonin strongly inhibited the melanin production of Streptomyces bikiniensis and B16 melanoma cells in comparison with a known melanogenesis inhibitor, arbutin.