A capillary zone electrophoresis (CZE) method, with UV detection and in the presence of dimethyl-beta-CD, was optimized by means of an experimental design for the separation and the simultaneous quantitation of ephedrine, pseudoephedrine, norephedrine (phenylpropanolamine) and norpseudoephedrine (cathine) in urine without any extraction. In this application, the optimization of the analytical conditions with an experimental design was preferred to a univariate study. Therefore, a central composite design was used and the following factors were investigated and varied simultaneously: buffer concentration, buffer pH and dimethyl-beta-CD concentration. In order to evaluate the influence of each experimental parameter on the analytical separation, the resolutions between the four compounds, as well as the separation time and generated current were observed and established as responses of the experimental design. A model was obtained for each response by linear multiple regression of a second-degree mathematical expression. After acceptance of the mathematical models, the most favorable conditions were determined by maximizing the resolutions between the four compounds and by setting the other responses at threshold values. Successful results were obtained with a 260 mM Tris-phosphate buffer at pH 3.5 in the presence of 13.3 mM dimethyl-beta-CD at 25degreesC and with an applied voltage of 30 W Under these optimized conditions, a baseline separation of the four compounds was achieved in less than 6 min. The method was validated in terms of precision, linearity, accuracy and successfully applied for the determination of these compounds in urine samples without any extraction as well as in nutritional supplements.