A New Enzymatic Method for the Determination of Free and Conjugated Glucuronic Acid

A New Enzymatic Method for the Determination of Free and Conjugated Glucuronic Acid Introduction Methods as yet available for the determination of glucuronic acid are based on the carbazole reaction ofDische (1). Various modifications of this colour reaction with higher sensitivity were summarized by Marsh (2). These tests have certain disadvantages; they are not specific, and they are subject to interference by various compounds, they are also time-consuming, and they do not differentiate between free and conjugated glucuronic acid. A less sensitive method using gas chromatography was described by Inoue (3). Until now, a specific enzymatic method for glucuronic acid determination was lacking. This paper describes a new method for the quantitative detenranation of glucuronic and galacturonic acid which is based on the NAEf-linked enzymatic oxidation of these uronic acids to the corresponding dicarboxylic acids with the stoichiometric formation of NADH: glucuronic acid uronic acid dehydrogenise galacturonic acid mucic acid glucaric acid NAPH tained are identical for both urpnic acids. In the presence of both uronic acids the galacturonic acid content can be determined according to Nagel (7) and subtracted from the total content determined by the uronic acid dehydrogenase test. The main interest of this study was the development of a sensititve glucuronic acid assay for samples containing only free http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Clinical Chemistry and Laboratory Medicine de Gruyter

A New Enzymatic Method for the Determination of Free and Conjugated Glucuronic Acid

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Publisher
de Gruyter
Copyright
Copyright © 2009 Walter de Gruyter
ISSN
1434-6621
eISSN
1437-4331
DOI
10.1515/cclm.1976.14.1-12.225
Publisher site
See Article on Publisher Site

Abstract

Introduction Methods as yet available for the determination of glucuronic acid are based on the carbazole reaction ofDische (1). Various modifications of this colour reaction with higher sensitivity were summarized by Marsh (2). These tests have certain disadvantages; they are not specific, and they are subject to interference by various compounds, they are also time-consuming, and they do not differentiate between free and conjugated glucuronic acid. A less sensitive method using gas chromatography was described by Inoue (3). Until now, a specific enzymatic method for glucuronic acid determination was lacking. This paper describes a new method for the quantitative detenranation of glucuronic and galacturonic acid which is based on the NAEf-linked enzymatic oxidation of these uronic acids to the corresponding dicarboxylic acids with the stoichiometric formation of NADH: glucuronic acid uronic acid dehydrogenise galacturonic acid mucic acid glucaric acid NAPH tained are identical for both urpnic acids. In the presence of both uronic acids the galacturonic acid content can be determined according to Nagel (7) and subtracted from the total content determined by the uronic acid dehydrogenase test. The main interest of this study was the development of a sensititve glucuronic acid assay for samples containing only free

Journal

Clinical Chemistry and Laboratory Medicinede Gruyter

Published: Jan 1, 1976

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