Objective To establish a method for the determination of aristolochic acid A in Jiuwei-Qianghuo pill. Method The HPLC system consisted of the Phenomenex Luna C18(4.6 mm × 250 mm, 5μm)column, the mobile phase consisted of acetonitrile and 0.05% H3PO4, gradient elution flow rate was 1.0 ml/min, the column temperature was 35℃, the UV detector was set at 250 nm.Results The linear response range was 0.014-0.280μg/ml(r=0.999 9). The detection limit and quantitation limit of aristolochic acid A in Jiuwei-Qianghuo pill was 1.5 and 4.5 ng/ml. The average recovery of aristolochic acid A was 96.3%. Conclusions The method was high sensitivity, accurate, repeatable and high secificity. It can be used as an inspection method for safe use of Jiuwei-Qianghuo pill.