Abstract: A capillary electrophoresis (CE) coupling with chemiluminescence (CL) detection method for determining hexestrol, dienestrol and diethylstilbestrol was developed based on the enhanced CL intensity of the luminol and Ag(Ⅲ) in alkaline solution. Based on the fact that hexestrol, dienestrol and diethylstilbestrol can dramatically restrain the chemiluminescence intensity of Luminol-Ag(Ⅲ) complex chemiluminescence system, a good linear relationship between the chemiluminescence intensity and concentrations of analytes is develped. A good baseline separation of hexestrol, dienestrol and diethylstilbestrol is obtained within 15 min. The calibration curves are linear in the range of 0.6~60 mg/L for Hexestrol and 0.5~40 mg/L for diethylstilbestrol and 1~60 mg/L for dienestrol. The limits of detection of dexestrol, diethylstilbestrol and dienestrol are 0.57 mg/L, 0.35 mg/L and 0.80 mg/L, respectively, with the relative standard deviation (RSD) of 2.38%, 1.90% and 1.3%. The method can be applied to the separation and the detection of milk sample. The proposed method was applied to detect hexestrol, dienestrol and diethylstilbestrol in milk sample with satisfactory assay results.