The chiral separation of amphetamines is of great importance in clinical and forensic analysis, because it is well known that enantiomers do not have identical pharmacological activity; for example, d-amphetamine is 3–4 times more effective in central nervous system stimulation than the l-isomer, whereas the latter is slightly more potent in its cardiovascular action. This paper describes the method development for the chiral separation of a mixture of amphetamine analogues by cyclodextrin-modified capillary zone electrophoresis. The use of different cyclodextrin types as chiral selectors and the influence of experimental parameters such as the temperature, the voltage applied, the buffer concentration and the capillary length are investigated. The ability to determined the enantiomers in urine samples is also discussed.