Boehme C L, Strobel H W
Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, 77225, USA.
J Chromatogr B Biomed Sci Appl. 1998 Nov 6;718(2):259-66. doi: 10.1016/s0378-4347(98)00368-5.
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.
已开发出灵敏的高效液相色谱（HPLC）方法，用于分离和定量氟哌啶醇及氯丙嗪代谢产物，以便在体外与纯化的细胞色素P450同工酶进行重组测定。使用Hypersil CPS柱，流动相为67%乙腈和10 mM醋酸铵（pH 5.4），实现了氟哌啶醇代谢产物的分离。使用Ultrasphere氰基柱，流动相为87.5%乙腈、5%甲醇、3% 0.12 M醋酸钠、3% 0.12 M醋酸铵、0.01%二乙胺和0.01%三乙胺（pH 9.5），实现了氯丙嗪代谢产物的分离。观察到氟哌啶醇及其三种主要代谢产物以及氯丙嗪及其五种主要代谢产物的分离效果良好。在重组系统中，母体化合物与纯化的细胞色素P450同工酶1A1和2B1进行体外孵育时，会形成不同水平和组合的这些代谢产物。
