Abstract: HPLC was applied to the determination of 5 sexual hormones in cosmetic. The cosmetic sample was extracted with methanol. Kromasil 100-5C18 chromatographic column was used as stationary phase, and a mixture of acetonitile and water (65+35) was used as mobile phase. UV-detection at 242 nm and 290 nm was adopted in the determination. Linear relationships between values of peak area and mass concentration of 5 sexual hormones were kept in the same range of 0.100-10.0 mg·L-1, with detection limits (3S/N) in the range of 0.552-1.01 mg·kg-1 and lower limits of determination (10S/N) in the range of 1.10-5.04 mg·kg-1. Values of recovery found by standard addition method were in the range of 96.2%-110%, and values of RSD′s (n=9) less than 10%.