Objective To establish a method for determination of Icariin in Yishengling granules (Sucrose-free) by HPLC. Methods Using a column packed with C18 and mixture of acetonitrile-water (30∶70) as the mobile phase. The flow rate was 1.0 mL/min and the temperature was 30 ℃. The detection wavelength was at 270 nm. Result The linear range of Icariin was 4.1～20.5 ?g (r=1). The mean recovery was 102.90% and RSD was 1.72%. Conclusion The method is simple, reliable and rapid, and can be applied to the quality control of Yishenling granules (Sucrose-free).