RP-HPLC method was applied for the assay of miconazole nitrate，dexamethasone acetate and paeonol in Mi Song Fen cream after the cream was pretreated with supersonic wave and ice bath. The linear range of miconazole nitrate was 40.5~648.0 μg/mL and r=0.999 1; the linear range of dexamethasone acetate was 3.05~48.8 μg/mL and r=0.999 8; the linear range of paeonol was 20.5~328.0 μg/mL and r=0.999 4. The mean recovery of miconazole nitrate, dexamethasone acetate and paeonol was 100.44±1.25 (%), 98.46±1.04 (%) and 98.24±1.29 (%), respectively. The present method was successful in removing interference substance in the cream and getting high recovery of the three drugs mentioned above，which will provide guidance for the quality control of cream.