Abstract
A method is described for the assay of UDP-glucuronyltransferase activity for various aglycones using UDP-[U 14-C]glucuronic acid. The radioactive glucuronide conjugates formed during the reactions were separated from unmetabolized UDP-[U- 14C]glucuronic acid, and from other uncharacterized radioactive products, by thin-layer chromatography. The conjugates were identified by comparison of their R f values on thin-layer chromatography with those of several authentic glucuronides, and by hydrolysis with β-glucuronidase in the absence or presence of d-saccharic acid-1,4-lactone, an inhibitor of β-glucuronidase. The procedure is general for the determination of the enzymic activity with various aglycones. It has been utilized for eleven aglycones and is capable of extension to many others.
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