A universal radiochemical high-performance liquid chromatographic assay for the determination of UDP-glucuronosyltransferase activity

被引:51
作者
Ethell, BT
Anderson, GD
Beaumont, K
Rance, DJ
Burchell, B [1 ]
机构
[1] Ninewells Hosp & Med Sch, Dept Mol & Cellular Pathol, Dundee DD1 9SY, Scotland
[2] Univ Washington, Sch Pharm, Seattle, WA 98195 USA
[3] Pfizer Cent Res, Dept Pharmaceut Res & Dev, Sandwich CT13 9NJ, Kent, England
基金
英国惠康基金;
关键词
D O I
10.1006/abio.1997.2443
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A new unified assay for the determination of UDP-glucuronosyltransferase (UGT) activities has been developed. The resolution of [C-14]uridine diphosphate glucuronic acid from radiolabeled glucuronides formed by incorporation of this radiolabel can now be achieved by a sensitive and rapid-gradient HPLC method which utilizes a radioactivity endpoint as a universal detection method. One important application of this method is the determination of kinetic parameters for cloned and expressed UGT isoforms with greater speed and precision than can be afforded by TLC methodology. Moreover, assays with C-14-labeled substrates indicate that gradient HPLC can easily resolve the substrate from the glucuronide products and present an alternative to the time-consuming optimization of conditions for organic phase extraction assays. (C) 1998 Academic Press.
引用
收藏
页码:142 / 147
页数:6
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