Aesculetin is the product of the hydrolysis reaction of aesculin. A high sensitivity and good repeatability method based on capillary electrophoresis with amperometric detection (CE-AD) was developed for simultaneous determination of aesculin and aesculetin in the hydrolysate of aesculin. Under the optimum condition: 10mmol/L KH(2)PO(4)-5mmol/L Na(2)B(4)O(7) (pH 6.0) buffer, separation at 18kV and +900mV (versus Ag/AgCl) as the detection potential, the hydrolysis rate constants of aesculin hydrolysis at 25, 30, 35, 40 and 45 degrees C in 0.1mol/L KOH were obtained as 1.45x10(-2)min(-1), 2.01x10(-2)min(-1), 2.93x10(-2)min(-1), 3.76x10(-2)min(-1) and 5.05x10(-2)min(-1), respectively. It was calculated that the activation energy for aesculin hydrolysis was 49.4kJ/mol.