Community Research and Development Information Service - CORDIS

A universal radiochemical high-performance liquid chromatography assay for the determination of UDP-Glucuronosyltransferase activity

A new unified assay for the determination of UDP-glucuronosyltransferase (UGT) activities was developed, using radio-labelled UDPGA and High-Performance Liquid Chromatography. 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-labelled substrates indicate that gradient HPLC can easily resolve the substrate from the glucuronide products and present an alternative to the time-consuming optimisation of conditions for organic phase extraction assays.

Reported by

University of Dundee
Ninewells Hospital & Medical School
DD1 9SY Dundee
United Kingdom
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