title: High-performance liquid chromatographic methods for the analysis of haloperidol and chlorpromazine metabolism in vitro by purified cytochrome P450 isoforms.
abstract: Sensitive HPLC methods for the resolution and quantitation of metabolites of both haloperidol and chlorpromazine metabolism have been developed for use in in vitro reconstitution assays with purified P450 isoforms. Separation of haloperidol metabolites was accomplished using a Hypersil CPS column with a mobile phase of 67% acetonitrile and 10 mM ammonium acetate, pH 5.4. Separation of chlorpromazine metabolites was achieved using an Ultrasphere cyano column with a mobile phase of 87.5% acetonitrile, 5% methanol, 3% 0.12 M sodium acetate, 3% 0.12 M ammonium acetate, 0.01% diethylamine and 0.01% triethylamine, pH 9.5. Sharp resolution was observed for haloperidol and three of its major metabolites and for chlorpromazine and five of its major metabolites. Varying levels and combinations of these metabolites are formed during in vitro incubations of parent compound with purified P450 isoforms 1A1 and 2B1 in a reconstituted system.
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, 77225, USA.
authors.forename: C L
authors.initials: CL
authors.lastname: Boehme
authors.forename: H W
authors.initials: HW
authors.lastname: Strobel
mesh_terms: D000818:Animals; D014150:Antipsychotic Agents / Q000378:metabolism; D002746:Chlorpromazine / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism*; D019362:Cytochrome P-450 CYP2B1 / Q000378:metabolism*; D006220:Haloperidol / Q000378:metabolism*; D051381:Rats
chemical_list: D014150:Antipsychotic Agents; D019363:Cytochrome P-450 CYP1A1; D019362:Cytochrome P-450 CYP2B1; D006220:Haloperidol; D002746:Chlorpromazine