Abstract: A method of HPLC for the determination of rutin in gardenia with solid phase extraction was proposed. The sample was extracted with a mixture of methanol and water (80+20) and purified by passing through C18-solid phase extraction column. C18 chromatographic column was used as stationary phase, and a mixture of acetonitile, 0.2 mmol·L-1 CuSO4 solution , triethylamine and acetic acid (30+70+0.5+0.7) was used as mobile phase. UV-detection at 360 nm was adopted in the determination. Linear relationship between values of peak area and mass concentration of rutin was kept in the range of 4.65-46.5 mg·L-1, with detection limit (3S/N) of 0.86 mg·L-1. Values of recovery found by standard addition method were in the range of 95.6%-97.5%, and values of RSD′s (n=6) less than 1.5%.