Application of Liquid Chromatography to a Study on 4-Coumarate: Coenzyme A Ligase Activity
Open Access
- 1 April 1981
- journal article
- research article
- Published by Walter de Gruyter GmbH in Zeitschrift für Naturforschung C
- Vol. 36 (3-4), 197-199
- https://doi.org/10.1515/znc-1981-3-402
Abstract
The separation of components from a 4-coumarate:CoA ligase assay by means of liquid chromatography was described. With the aid of polyamide column chromatography it is possible to enrich and isolate chromatographically and UV spectroscopically pure p-coumaroyl-CoA using as a solvent 0.01% NH4OH in methanol subsequent to water and methanol alone. High performance liquid chromatography (HPLC) on octadecysilane-bonded silica stationary phase allows a discontinous determination of ligase activity. All components (ATP, Coenzyme A, p-coumaric acid and the products AMP and p-coumaroyl -CoA) and be separated and accurately quantified within 20 min using a water-acetonitrile gradient, containing 1% phosphoric acid. The HPLC method may be used to affirm the accuracy of optical tests.Keywords
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