Abstract: OBJECTIVE An HPLC method was established for the simultaneous determination of Chlorogenic Acid, Rutin, Isoquercitrin and Quercetin in drug Mulberry Leaves of Xinjiang. METHODS The chromatographic separation was carried out on a Yilite Hypersil BDS (4.6 mm × 250 mm, 5 μm) with a mobile phase composed of acetonitrile (A) and 0.01 mol·L^-1 ammonium acetate(pH=4.8) (B) with gradient elution 0 min, A-B (8∶92)；30 min, A- B (30∶70) . The flow rate was 1.0 mL·min^-1, the detection wavelength was set at 350 nm and the column temperature was 25 oC. RESULTS The linear ranges of Chlorogenic acid , Rutin, Isoquercitrin, and Quercetin were between 1.052 0-5.067 9 μg(r=0.999 3), 1.029 4-5.031 7 μg (r=0.999 8), 1.076 8-5.065 5 μg (r=0.999 0) and 1.075 0-5.055 0 μg (r=0.999 1), respectively. The average recoveries of Chlorogenic Acid, Rutin, Isoquercitrin and Quercetin were 102.2% (RSD=1.3%), 104.6% (RSD=0.9%), 104.0% (RSD=1.9%), 104.7% (RSD=1.2%). The detection limits were 0.160, 0.081, 0.033, 0.640 μg·mL^-1(S/N=3), respectively. CONCLUSION The method is simple, rapid, sensitive and accurate. Therefore it is applicable for the determination of Chlorogenic Acid, Rutin, Isoquercitrin and Quercetin in drug Mulberry Leaves of Xinjiang.