Objective To develop an HPLC method for simultaneous determination of daphne glycoside, daphnetin-8-O-β-Dglucoside, daphnetin, and 7-hydroxycoumarin in Zushima Gel Ointment.Methods The HPLC method was used on Prodigy ODS(3) column(250 mm x 4.60 mm, 5 μm).The mobile phase was consisted of methanol-0.05% phosphoric acid(18:82).The detection wavelength was set at 327 nm.The temperature of column was set at 35 °C, the flow rate was 1.0 mL/min, and the volume of injection was 10 μL.Results The linear ranges of daphne glycoside, daphnetin-8-O-β-D-glucoside, daphnetin, and 7-hydroxycoumarin were 0.105 7-1.268 4 μg, 0.092 2-1.106 4 μg, 0.060 5-0.726 0 μg, and 0.050 5-0.606 0 μg, resp., with r values above 0.999 9.The average recoveries were 97.22%, 99.27%, 98.78%, and 99.45% with RSD values of 1.01%, 1.10%, 1.40%, and 1.67%, resp.Conclusion The method is simple, accurate, sensitive, and reproducible, and at the same time, it provided data support for establishing a reliable quality standard of Zushima Gel Ointment, and controlling the quality of the preparations better and more comprehensively.