The aim of this paper is to determine the berberine hydrochloride in kangjixiaoke tablet by HPLC. The HPLC system consisted of Ailtima C18 column (250 mm × 4.6 mm, 5 μm) with acetonitrile-0.033 mol/L-1-KH2PO4 (30:70) as mobile phase and UV detector at wavelength of 265 nm. The result showed that the berberine curves were linear in the range of 0.10-0.80 μg (r=0.999 9). The regression equation was Y = 71.41C -16.125 and the precision test RSD was 0.65%. The mean recovery was 101.2% and RSD was 1.4%. It was concluded that the method is simple, accurate, and reliable for determination of berberine in kangjixiaoke tablet.