OBJECTIVE:To establish a method for simultaneous determination of the confents of calycosin-7-glucoside,calycosin and formononetin in Shenqi granule. METHODS:HPLC was performed on the column of Welch Ultimate XB-C18 with mobile phase of acetonitrile-0.2% Formic acid solution(gradient elution)at flow rate of 1.0 ml/min,detection wavelength was 250 nm, column temperature was 35 ℃,and injection volume was 10 μl. RESULTS:The linear range of sample quantity was 0.008-0.32 μg for calycosin-7-glucoside(r=0.999 9),0.005-0.20 μg for calycosin(r=0.999 7)and 0.007-0.28 μg for formononetin(r=0.999 8); RSDs of precsion,stability and reproducibility tests were no more than 1.59%;recoveries were 98.17%-99.17%(RSD=0.67%, n=6),98.03%-99.66%(RSD=0.57%,n=6)and 98.16%-99.12%(RSD=0.43%,n=6). CONCLUSIONS:The method is simple, accurate and reliable,and can be used for the content determination of Shenqi granule ofiso flavones.