OBJECTIVE: To establish a method for simultaneous determination of chlorogenic acid, baicalin and imperatorin in Biyanling tablets. METHODS: HPLC method was adopted. The separation was performed on a Waters SunFire C18 column with mobile phase consisted of methanol-0.20% phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 280 nm and column temperature was 30 ℃. The sample size was 20 μL. RESULTS: Linear ranges of chlorogenic acid, baicalin and imperatorin were 1.618-16.18 μg/mL, 1.624-16.24 μg/mL and 1.762-17.62 μg/mL (r were 0.999 9), respectively. The limits of quantitation were 2.67,2.03,1.87 μg/mL, and the limits of detection were 1.34, 1.06, 0.98 μg/mL, respectively. RSDs of precision, stability and reproducibility tests were all lower than 2.0%. The recoveries were 98.01%-101.80%(RSD=1.38%,n=9), 98.38%-101.40%(RSD=1.26%,n=9)and 98.06%-101.40%(RSD=1.28%,n=9), respectively. RSDs of tolerance tests were all lower than 2.0%. CONCLUSIONS: The method is simple, accurate, precise, stable, reproducible and durable. It can be used for simultaneous determination of hlorogenic acid, baicalin and imperatorin in Biyanling tablets.