Experimentally determined optimum separation conditions for a metabolite of anaesthetic halothane in urine, trifluoroacetic acid, were assessed by means of computer simulation of the isotachophoretic steady-state. The simulation confirmed that urinary acids and trifluoroacetic acid can be separated in the limited pH range of 3.5-3.7 buffered by beta-alanine, as far as the pH dependence of effective mobility is utilized. The separated fraction of the trifluoroacetic acid zone was identified by mass spectrometry. The simulated coefficient of the calibration curve agreed well with the observed value.