Abstract: Objective:A new capillary zone electrophoresis (CZE) method was developed for the simultaneous quantification of rutin, kaempferol and quercetin in Celastrus orbiculatus Thunb. Different results were analysed and the medicinal quality control methods were discussed. Methods:The electrophoretic analysis was performed on 30mmol·L-1 sodium borate solution (pH=9.0) , 20kV electrophoresis voltage and 254nm wavelength. Results:Rutin, kaempferol and quercetin were separated successfully in 12min by CZE, a good linearity between peak area and concentration was found in range of 2.5⁴00μg·mL-1 for three compounds. Conclusion:Flavonoids were abundant in Celastrus orbiculatus Thunb. Simultaneous determination of rutin, kaempferol and quercetin by CZE method had a good accuracy and precision. The method was simple, fast and applicable for clinical and scientific research to determine three flavonoids. This provided a technology method and solid theoretical data for Celastrus orbiculatus Thunb. of medicinal quality control.