A capillary zone electrophoresis method is described for the simultaneous determination of atropine, homatropine and scopolamine. Successful results were obtained after optimization of the electrophoretic parameters such as buffer composition and pH. The best separation was achieved using a 100 mM Tris-phosphate running buffer at pH 7. The validation data proved that the method had the requisite selectivity, sensitivity, reproducibility and linearity to be used for the assay of these compounds in pharmaceutical formulations. Dosage of the separate drugs in ophthalmic preparations is also presented.