OBJECTIVE: To establish a method for simultaneous determination of paeoniflorin and salvianolic acid B in Ruangan suopi pill. METHODS: HPLC was performed on the column of Wondasil C18 with mobile phase of acetonitrile -0.1% phosphoric acid solution (gradient elution) at a flow rate of 1.0 mL/min, the detection wavelength was 230 nm for paeoniflorin and 286 nm for salvianolic acid B, the column temperature was 30 ℃, and the injection volume was 10 μL. RESULTS:The linear range was 9.79-195.80 μg/mL(r=0.999 9) for paeoniflorin, 11.45-229.00 μg/mL(r=0.999 9) for salvianolic acid B; the limits of quantification were 0.018 1,0.014 4 μg, limits of detection were 0.005 4, 0.004 3 μg; RSDs of precision, stability and reproducibility tests were lower than 2.0%; the recoveries were 98.67%-102.22%(RSD=1.26%,n=9) and 99.72%-103.28%(RSD=1.05%,n=9). CONCLUSIONS: The method is rapid, sensitive and accurate, and can be used for the simultaneous determination of paeoniflorin and salvianolic acid B in Ruangan suopi pill.