Abstract
A method is described for the determination of L-carnitine in the range 20–2000 pmoles. Incubation of L-carnitine with [1- 14C]acetyl-coenzyme A and acetyl-CoA: carnitine O-acetyltransferase (EC 2.3.1.7) yields labeled acetylcarnitine. The formed acetylcarnitine is separated from labeled acetyl-coenzyme A by passing the reaction mixture through a column of the anion exchange resin Dowex 2-X8 and its isotope content determined in a liquid scintillation counter. Short-chain acyl derivatives of L-carnitine are substrates for the enzyme and will also be determined through exchange in the enzymic reaction. In 33 samples of human blood plasma the concentration of carnitine (+short-chain acyl carnitine) was 51 ± 10.5 μmoles/ l (mean ± S.D.). In biopsies of human skeletal muscle obtained at surgery the concentration was 12.8 ± 2.9 μmoles/g dry weight, in mouse skeletal muscle 1.73 ± 0.27 μmoles/g dry weight, and in mouse heart 4.66 ± 0.73 μmoles/g dry weight. The precision of the method expressed as relative standard deviation of duplicate analyses was 2–5%. The recovery of carnitine added to plasma or to perchloric acid extracts of muscle was 97 ± 12.5% and 103 ± 5.9%, respectively.
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