A reversed-phase high-performance liquid chromatographic (HPLC) method for the analysis of perphenazine in rabbit plasma was developed. Amitriptyline HCl was used as the internal standard. The chromatography was performed using a Microsorb-CN column; the mobile phase consisted of 85:15 methanol/0.005 M ammonium acetate buffer; flow rate of 1.6 ml/min; and ultraviolet detection at 254 nm. Retention times were 10.8 and 18.1 min for perphenazine and amitriptyline HCl, respectively. The recovery of perphenazine added to plasma in the concentration range of 0.05-5 micrograms/ml was found to be greater than 80%. This procedure was used for the analysis of plasma samples collected over time following the oral administration of perphenazine tablets in rabbits.