Direct measurement of free coenzyme A in biological extracts by reversed-phase high-performance liquid chromatography.
A high-performance liquid chromatographic system was developed with baseline separation of coenzyme A (CoASH) from dephospho-coenzyme A and acetyl-coenzyme A using isocratic elution. The chromatographic separation was achieved in a reversed-phase system with a high concentration (220 mM) of potassium phosphate buffer at pH 4.0 and appropriate amounts of methanol (ca. 12%). The eluate was monitored with a UV detector at 254 nm with the limit of detection at ca. 5 pmoles. The system could be used without modification for the estimation of the content of CoASH in biological extracts, e.g. mitochondria.
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003065:Coenzyme A / Q000032:analysis*; D008930:Mitochondria, Liver / Q000201:enzymology; D051381:Rats
D003065:Coenzyme A