Measurement of free and esterified carnitine in tissue extracts by high-performance liquid chromatography.
A high-performance liquid chromatographic (HPLC) method for the determination of L-carnitine in clamped and frozen rat livers is described. L-carnitine + acetyl-CoA in equilibrium with acetyl-L-carnitine + CoASH Using the above enzymatic reaction, release of CoASH is stoichiometric with the L-carnitine added. The present method has made possible the determination of carnitine in liver tissues, which is difficult by the conventional enzymatic spectrophotometric method using 5,5'-dithiobis(2-nitrobenzoic acid), owing to acetyl-CoA hydrolysis during prolonged incubations at pH 7.8.
D000105:Acetyl Coenzyme A / Q000032:analysis; D000214:Acyl Coenzyme A / Q000032:analysis; D000818:Animals; D002331:Carnitine / Q000032:analysis*; D002332:Carnitine O-Acetyltransferase / Q000378:metabolism; D003065:Coenzyme A / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000032:analysis*; D008623:Mercaptoethanol / Q000378:metabolism; D051381:Rats
D000214:Acyl Coenzyme A; D008623:Mercaptoethanol; D000105:Acetyl Coenzyme A; D002332:Carnitine O-Acetyltransferase; D002331:Carnitine; D003065:Coenzyme A