To establish the HPLC method to determine content of berberine hydrochloride in Berberine Tablets,and to ensure the results are accurate and reliable by systematical testing methodology.Methods：The chromatographic column：Agilent TC-C18 column (4.6×150 mm,5 μ m,Agilent,America),mobile phase：acetonitrile-0.05 mol/L potassium dihydrogen phosphate (0.05 mol/L sodium heptanesulfonate solution 45∶55,5%),the detection wavelength was 265nm.Results：The berberine hydrochloride in 2.5ug·mL-1-80ug·mL-1 linear relationship was good,and the linear regression equation was Y = 421.5X+17.32,and correlation coefficient was 0.9997 (n = 6); low,high concentration within batch precision values of RSD were 0.30%,0.58%,0.30%,inter batch precision RSD was 0.80%; repeatability RSD was 0.27%; the low,recoveries,in high concentrations were 99.66%,99.88%,99.98%; the test solution was placed at room temperature for 8h,and RSD was 0.41%.Conclusion：This method for determination of berberine hydrochloride in tablet of berberine is accurate and reliable with simple operation and short time.It can be used for content determination of berberine hydrochloride in tablets of berberine.