Objective: To establish a method for the determination of the content of berberine in LianBo burn ointment.Methods: HPLC method was used.Hypersil ODS2 C18 (150 mm × 406 mm, 5 μm) was used as chromatog. column.Mobile phase was 0.033 mol/L Potassium dihydrogen phosphate solution-acetonitrile (60:40).The detection wavelength was 265 nm.Results: Berberine hydrochloride had a good linearship between 0.11∼0.33 μxg (r = 0.9996).The average recovery was 99.51%.RSD was 0.99%.Conclusion: The method is simple, accurate, good reproducible and detection of low cost.It can be used for the quality control of Lianbo Bum ointment.