A simple,low-cost and effective reverse flow-high performance capillary electrophoresis(RF-HPCE)method was developed for the separation and determination of tenuifolin in Polygalae Radix.Through optimization of separation conditions, a buffer of 50 mmol/L borax containing methanol(65:35,v/v)was selected for the separation of analytes.Regression equation revealed good linear relationship between peak area and concentration of the analyte in the range of 0.0563-0.9000 mg/mL (r=0.9988).The recoveries ranged from 99.50%-104.7%.The results indicated that this method is simple,rapid and accurate. It can be used for the quality control of Polygalae Radix.