The inhibitive components in anti-acne cosmetics including spironolactone, benzoyl peroxide, and tretinoin were simultaneously determined by reversed-phase high performance liquid chromatography (RP-HPLC). The cosmetics were extracted with methanol by microwave and analyzed by high performance liquid chromatography. The HPLC conditions were as follows: Kromasil C18 column (4.6 mm i. d. x 250 mm, 5 microm), methanol and phosphate buffer as mobile phase with gradient elution at a flow rate of 1.0 mL/min, UV detection at 265 nm. Three components were separated completely within 11 min. The calibration curves of the three compounds were linear (nu > 0.999 9) between 1 and 200 mg/L. The average recoveries were from 88.2% to 106.7% with relative standard deviations lower than 3. 1%. The detection limits (S/N = 3) were 0.101 mmicrog for spironolactone, 0.100 microg for benzoyl peroxide, and 0. 107 microg for tretinoin. The method is simple and rapid with high accuracy, and suitable for the determination of the 3 inhibitive components in anti-acne cosmetics.