Application of liquid chromatography to a study on 4-coumarate: coenzyme A ligase activity.
نویسندگان
چکیده
This report describes the separation of components from a 4-coumarate:CoA ligase assay by means of liquid chromatography. 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 on octadecylsilane-bonded silica stationary phase allows a discontinuous determination of ligase activity. All components - ATP, Coenzyme A, p-coumaric acid, and the products AMP and p-coumaroyl-CoA - can be separated and accurately quantified within 20 min using a water-acetonitrile gradient, containing 1% phosphoric acid. The presented HPLC method may be used to affirm the accuracy of optical tests.
منابع مشابه
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عنوان ژورنال:
- Zeitschrift fur Naturforschung. Section C, Biosciences
دوره 36 3-4 شماره
صفحات -
تاریخ انتشار 1981