Objective To develop an RP-HPLC method for the content determination of heseridin in Hezhong Lipi Pills.Methods The Agilent HC-C18 column (250 mm×4.6 mm, 5μm) was used with methanol-0.1% phosphoric acid (35∶65) as the mobile phase at the flow rate of 1.0 mL/min. The detection wavelength was 283 nm, and the column temperarute was 40℃.Results The linear range of hesperidin was 0.15-3.64μg,r=1.000 (n=6). The average recovery was 98.97%, RSD=0.90% (n=6).Conclusion The method is accurate, simple, sensitive and with good repeatability, which can be applied to the quality control ofHezhong Lipi Pills.