Abstract: The metabolites of perphenazine in human bile were rapid investigated employing UPLC/Q-TOF MS^E (where E represents energy) technique. The bile samples were collected after dosing to human and purified through acetonitrile protein precipitation. Data collection was done by UPLC/Q-TOF MS^E. According to the molecule formular educed from the measured accurate mass, mass defect filter(MDF) technique and generic dealkylation tool were used to screen the metabolites. Their structures were elicited by comparing the fragmentation patterns between parent drug and metabolites. All the metabolites has chlorine atom isotope cluster. After oral administration of perphenazine to the patient, 29 metabolites including 16 phase I and 13 phase II metabolites are detected in human bile. The major metabolic pathways of perphenazine in human are hydroxylation, N-dealkylation, desaturation, sulfation and glucuronate conjugation.