OBJECTIVE: To establish a method for the simultaneous determination of paeoniflorin and paeonol in Dirda pill,and provide reference for improving its quality control standard. METHODS: HPLC was performed on the column of ZORBAX C18 with mobile phase of acetonitrile-0.1% phosphoric acid (gradient elutio) at a flow rate of 0.8 ml/min, detection wavelength was 230 nm(paeoniflorin), 274 nm(paeonol), columne temperature was 35 ℃, injection volume was 20 μl. RESULTS: The linear range was 0.514 8-1.544 5 μg for paeoniflorin (r=0.999 2) and 0.643 2-1.960 4 μg for paeonol(r=0.999 8); the limits of quantification were 0.135 6, 0.126 4 μg, limits of detection were 0.067 8, 0.063 2 μg; RSDs of precision, stability and reproducibility tests were lower than 2%; recoveries were 95.78%-97.27%(RSD=0.62%,n=6) and 97.38%-98.38% (RSD=0.42%,n=6). CONCLUSIONS: The method is simple with good reprosucibility and high sensibility, and can be used for the simultaneous determination of paeoniflorin and paeonol in Dirda pill.