The related substances in Butenafine Hydrochloride Cream were determined A C18 column was used with the mobile phase of acetate buffer-methanol-isopropanol (17:70:13). The detection wavelength was 282 nm. The flow rate was 1.0 mL/min. The calibration curve was linear in the range of 0.625-5.000 μg/mL for butenafine hydrochloride (r = 0.9989). The detection limit was 2.6 ng. The method was good in the aspects of separation, sensitivity and repeatability, and could be used for the quality control of related substances of butenafine hydrochloride.