High-performance liquid chromatography of imipramine and six metabolites in human plasma and urine.

A method for the simultaneous quantitation of imipramine and six metabolites (2- and 10-hydroxyimipramine, 2- and 10-hydroxydesipramine, didesmethylimipramine and desipramine) in human plasma and urine has been developed. The method is based on a three-step liquid-liquid extraction followed by isocratic, reversed-phase high-performance liquid chromatography with ultraviolet absorbance detection (detection wavelength: 220 nm). The chromatographic eluent consisted of 30% acetonitrile and 70% aqueous sodium perchlorate solution pH 2.5. Glucuronide conjugates in urine were deconjugated with beta-glucuronidase/arylsulphatase prior to extraction.