Abstract: OBJECTIVE To establish an accurate, universal and efficient HPLC method for simultaneous determitation of 16 glucocorticoids illegally added in Chinese patent medicine and health products. METHODS HPLC method was performed on an Agilent Extend-C₁₈(250 mm×4.6 mm, 5 μm) column with the mobile phase consisted of 0.1 mol·L^-1 ammonium?acetate solution-acetonitrile in a gradient elution mode. The detector wavelength was set at 240 nm. RESULTS The linear range of 16 glucocorticoids were 0.2-25 μg·mL^-1, which showed a good correlation. The recoveries were good except plaster(≥78.5%), except the emplas trum with the recovery rate of 57.2%-83.3%, and the detection limits were all <0.05 ng·mL^-1. CONCLUSION The method is simple and efficient, accurate and universal. It can be used as an efficient method for determination of 16 commonly used glucocorticoids which are illegally added in Chinese patent medicine and health products.