Objective: The content of hesperidin in Jinshuiliujun Decoction (JSLJD) was determined by RP HPLC. Methods: The chromatographic column was Hyperisl BDS C 8(10?). The mobile phase was acetonitrile water containing 8% acetic acid (17∶83). The flow rate was 1.0 mL/min. The column temperature was 40 ?C and the detection wavelength was 284nm. The external standard and peak area method were used to determine hesperidin content. Results: There was a very good linear relationship between hesperidin and peak area when the sample injection was from 0.02 to 2.00?g. The average recovery of adding sample was 100.5% and RSD was 1.85%. Conclusions: This method is simple and believable. It can be effectively used for the quality control of JSLJD.