Objective To establish the method to determine the content of berberine in Double Yellow Antiinflammatory Tablets. Methods Using HPLC, with Diamonnsil C18 (4.6 mm?250 mm, 5 ?m) as column, acetonitrile-0.05 mol/L potassium dihydrogen phosphate (35∶65, phosphate regulation pH value to 3.0) as mobile phase, detection wavelength was at 345 nm, flow rate was 0.8 mL/min, and column temperature was 30 ℃. Results In berberine range of 0.040 7～0.447 9 ?g, the scope and content of the sample peak area had a good linear relationship, reproducibility of RSD=0.59% (n=5), and the average recovery rate was 96.71% and RSD=0.37% (n=5). Conclusion This method is stable, reliable, with high sensitivity and good reproducibility, and easy to put into practice, so it can be used for quality control of Double Yellow Antiinflammatory Tablets.