Lahoz Agustín, Donato Maria Teresa, Montero Sandra, Castell José V, Gómez-Lechón Maria José
Unidad Mixta Fundación Hospital La Fe-Advancell, Avenida Campanar 21, Valencia, Spain.
Rapid Commun Mass Spectrom. 2008;22(2):240-4. doi: 10.1002/rcm.3359.
Primary hepatocytes are still the best qualified in vitro system to anticipate drug metabolism in man. Recent advances in hepatocytes cryopreservation have notably increased their use not only for drug metabolism studies, but also for other applications such as cell transplantation. Evaluation of the drug-metabolizing competence of each hepatocytes preparation is needed. To date, the metabolic characterization of hepatocytes preparations relies on the assessment of phase I activities and the role of phase II enzymes receives little attention. A novel approach for the rapid assessment of the metabolic functionality of hepatocytes has been developed. A five-probe cocktail was used to simultaneously determine the enzymatic activities of major human phase I CYPs (CYP1A2, CYP2A6, CYP2C9, CYP2E1, CYP3A4), as well as two phase II enzymes: glucuronidase (UGTs) and sulfotransferase (SULT). Liquid chromatography/tandem mass spectrometry was used as the technique of choice for the determination of the enzymatic activities in a single run. Results showed that the method described herein permits a rapid assessment of the metabolic capabilities of human hepatocyte preparations as well as an estimation of the quality of freshly isolated or cryopreserved hepatocytes.
原代肝细胞仍是预测人体药物代谢的最佳体外系统。肝细胞冷冻保存技术的最新进展显著增加了其应用，不仅用于药物代谢研究，还用于细胞移植等其他应用。需要评估每种肝细胞制剂的药物代谢能力。迄今为止，肝细胞制剂的代谢特征依赖于对I相活性的评估，而II相酶的作用很少受到关注。已开发出一种快速评估肝细胞代谢功能的新方法。使用一种五探针混合物同时测定主要人I相细胞色素P450（CYP1A2、CYP2A6、CYP2C9、CYP2E1、CYP3A4）以及两种II相酶：葡萄糖醛酸酶（UGTs）和磺基转移酶（SULT）的酶活性。液相色谱/串联质谱法被用作单次运行中测定酶活性的首选技术。结果表明，本文所述方法能够快速评估人肝细胞制剂的代谢能力，并估计新鲜分离或冷冻保存的肝细胞的质量。
