Abstract: HPLC was applied to the determination of peoniflorin in traditional Chinese medicine and health food with solid phase extraction. The sample was extracted with 30%(φ) ethanol solution,and purified by passing through acidic Al2O3 solid phase extraction column. C18 chromatographic column was used as stationary phase with mixtures of acetonitrile and 0.1%(φ) H3PO4 solution with different mixing ratios as mobile phase in gradient elution. UV-detection at 230 nm was adopted in the determination. Linear relationship between values of peak area and mass concentration of peoniflorin was kept in the range of 2.00-200 mg·L-1,with detection limit (3S/N) of 0.5 mg·L-1. Values of recovery found by standard addition method were in the range of 99.5%-101%,and values of RSD′s (n=6) less than 2%.