OBJECTIVE：To establish a method for the content determination of hordenine in Zang medicine Herba Aconiti. METHODS：HPLC was performed on the column of Gemini-NX C18 with mobile phase of acetonitrile-0.25 mol/l Ammonium acetate solution（ammonia water to adjust the pH to 9.5，gradient elution，at flow rate of 1 ml/min，the detection wavelength was 230 nm，the column temperature was 30 ℃，and the injection volume was 20 μl.RESULTS：The linear range of hordenine was 3.276-819 μg/ml（r＝0.999 5）；RSDs of precision，stability and reproducibility tests were lower than 2.0％；recovery was 96.21％-104.04％ （RSD＝1.23％，n＝9）. CONCLUSIONS：The method is simple，stable and reproducible，and can be used for the content determination of hordenine in Zang medicine Herba Aconiti.