Abstract
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.
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References
Rebouche CJ (1992) Carnitine function and requirements during the life cycle. FASEB J 6(15):3379–3386
Jones PM, Bennett MJ (2012) Disorders of the carnitine cycle and mitochondrial fatty acid oxidation. In: Garg U, Smith LD, Heese BA (eds) Laboratory diagnosis of inherited metabolic diseases. AACC Press, Washington D.C., pp 65–91
Longo N, Amat di San Filippo C, Pasquali M (2006) Disorders of carnitine transport and the carnitine cycle. Am J Med Genet C Semin Med Genet 142C(2):77–85
Ho CS, Cheng BS, Lam CW (2003) Rapid liquid chromatography-electrospray tandem mass spectrometry method for serum free and total carnitine. Clin Chem 49(7):1189–1191
Minkler PE, Stoll MS, Ingalls ST, Kerner J, Hoppel CL (2015) Validated method for the quantification of free and total carnitine, butyrobetaine, and acylcarnitines in biological samples. Anal Chem 87(17):8994–9001
Morand R, Donzelli M, Haschke M, Krahenbuhl S (2013) Quantification of plasma carnitine and acylcarnitines by high-performance liquid chromatography-tandem mass spectrometry using online solid-phase extraction. Anal Bioanal Chem 405(27):8829–8836
Scott D, Heese B, Garg U (2016) Quantification of free carnitine and acylcarnitines in plasma or serum using HPLC/MS/MS. Methods Mol Biol 1378:11–19
Sowell J, Fuqua M, Wood T (2011) Quantification of total and free carnitine in human plasma by hydrophilic interaction liquid chromatography tandem mass spectrometry. J Chromatogr Sci 49(6):463–468
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Baird, S., Frazee, C.C., Garg, U. (2022). Quantification of Free and Total Carnitine in Serum Using Liquid Chromatography Tandem Mass Spectrometry. In: Garg, U. (eds) Clinical Applications of Mass Spectrometry in Biomolecular Analysis. Methods in Molecular Biology, vol 2546. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2565-1_9
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DOI: https://doi.org/10.1007/978-1-0716-2565-1_9
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