A method based on capillary electrophoresis with electrochemical detection has been developed for the separation and determination of paeoniflorin, sucrose, paeonoside, glucose, and fructose in Moutan Cortex for the first time. Effects of several important factors such as the concentration of NaOH, separation voltage, injection time, and detection potential were investigated to acquire the optimum conditions. The detection electrode was a 300 microm diameter copper disc electrode at a working potential of +0.60 V (versus saturated calomel electrode (SCE)). The five analytes can be well-separated within 12 min in a 40 cm length fused silica capillary at a separation voltage of 12 kV in a 75 mM NaOH aqueous solution. The relation between peak current and analyte concentration was linear over about 3 orders of magnitude with detection limits (S/N = 3) ranging from 0.9 to 1.3 microM for all analytes. The proposed method has been successfully applied to monitor glycoside and sugar contents in the real plant samples with satisfactory assay results.