Traditional Chinese medicine (TCM) has been frequently used as skin lightning agents. However, the mechanism of action of their effect is unclear. The present study aims to evaluate anti-tyrosinase activity of 10 commonly used TCM on mushroom (ab), human (hs) and mouse melanoma B16F0 (mm) tyrosinase (TYR) respectively. The results showed that at 1.0 mg/mL, extracts from Rosa rugosa Thumb, Morus alba L. and Paeonia lactiflora Pall were active against both abTYR and hsTYR (>50% inhibition), extracts from Bletilla striata (Thunb.) Rchb. F., Centella asiatica (L.) Urb, Cynanchum atratum L., Rosa canina L., Rhus chinensis Mill. and Glycyrrhiza urolensis Fisch. Ex DC. inhibited either abTYR or hsTYR (>50%), while extract from Tribulus terrestris L. had no/minimal activity (<10% inhibition). When treated with melanoma B16F0 cells, M. alba also significantly reduced mmTYR activity (70% at 250 μg/mL) and melanin content (50% at 250 μg/mL). These findings demonstrated inhibitory effects of 9 TCM against TYR and hence support their application as skin lightning agents. Our results also showed discrepancies in TYR activity from different sources, suggesting a testing regime of combining abTYR, hsTYR and mmTYR when developing depigmentation agents for human application.