Quantification of Free and Total Carnitine in Serum Using Liquid Chromatography Tandem Mass Spectrometry.
L-carnitine is a crucial component for transporting long-chained fatty acids from the cytosol into the mitochondrial matrix for fatty acid oxidation. During this process, carnitine forms numerous acylcarnitines before being recycled into the cytosol. Abnormal levels of free carnitine, total carnitine, and acylcarnitines in serum can be indicative of a metabolic disorder before symptoms are present. A liquid chromatography tandem mass spectrometry (LC-MS/MS) method is described for the determination of free and total carnitine in serum. To measure total carnitine, samples are spiked with deuterated carnitine (internal standard) and hydrolyzed with potassium hydroxide to convert acylcarnitines to carnitine. The reaction is quenched by the addition of hydrochloric acid. Carnitine is extracted via a methanolic protein precipitation. The solution is then injected on LC-MS/MS for analysis to determine the carnitine concentration using multiple-reaction monitoring.
Methods in molecular biology (Clifton, N.J.)
Carnitine; Chromatography, Liquid; Fatty Acids; Hydrochloric Acid; Tandem Mass Spectrometry
Acylcarnitine; Carnitine; Fatty acid oxidation; Tandem mass spectrometry
Baird S, Frazee CC 3rd, Garg U. Quantification of Free and Total Carnitine in Serum Using Liquid Chromatography Tandem Mass Spectrometry. Methods Mol Biol. 2022;2546:95-104. doi:10.1007/978-1-0716-2565-1_9