OBJECTIVE: To establish a method for the content determination of icariin in Kangguzhi zengsheng tablet,and provide a basis for improving the quality standard. METHODS: HPLC was performed on the column of Hypersil ODS2 C18 with mobile phase of acetonitrile - water (27 ∶ 73,V/V) at a flow rate of 1.0 ml/min, detection wavelength was 270 nm, column temperature was 25 ℃, and injection volume was 10 μl. RESULTS: The linear range of icariin was 3-60 μg/ml (r=0.999 9); limit of quantitation was 11.2 μg/ml, limit of detection 4.3 μg/ml; RSDs of precision, stability and reproducibility tests were lower than 2.0%; recovery was 99.00%-100.67%(RSD=0.63%, n=6). CONCLUSIONS: The method is simple, accurate and reproducible, and can be used for the content determination of icariin in Kangguzhi zengsheng tablet.