Abstract:Objective: To establish an HPLC method for the simultaneous determination of three major bioactive components in Jingzhi Guizhi Fuling capsules namely laetrile, paeoniflorin and paeonol. Method: A LiChrospher C18 column(4.6 mm×250 mm,5 μm) was used. The chrornatography was carried out with a stepwise gradient programming. The mobile phase was acetonitrile-water (containing 0.1% phosphorous acid ) and the flow rate was 1.0 mL·min1. Result: The linear range of laetrile was 12.87-102.94 μg·mL-1, r=0.999 9, paeoniflorin 24.84-198.7 (μg·mL-1,r = 0.9999 and paeonol 12.57-100.56 μg·mL-1,r= 0.999 9. The method is accurate with variation less than 1.5 % and recovery more than 95 % . Conclusion: The method was successfully applied to analyze three major bioactive components in Jingzhi Guizhi Fuling capsules.