Abstract:Objective: To establish a HPLC method for determination two constituents in bark of Paeonia Suffruticosa. Method: The reversed phase HPLC system consisting of an Alltima ODS column (4.6 mm ×150 mm,5 μm) and a mixture of water-THF-methanol-HAc (60:20:20:0.05) as the mobile phase was used. The flow rate was 0.8 mL·min-1 and UV detection was set at 274 nm. Result: The assay displayed good linearity over the concentration ranges of 0.06-1.0 μg (r=0.999 9, gallic acid) and 0.16-2.58 μg(r=0.9999, paeonol) respectively. The average recoveries (n=9) of gallic acid and paeonol were 98.6% (RSD=3.0%),98.2%(RSD=2.5%), respectively. The samples were extracted with methanol for 24 h bu maceration. Conclusion: The method is simple, accurate and can be used for the quality study of bark of P. suffruticosa.