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Clinical Chemistry 40: 817-821, 1994;
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Clinical Chemistry, Vol 40, 817-821, Copyright © 1994 by American Association for Clinical Chemistry

Carnitine determination by an enzymatic cycling method with carnitine dehydrogenase

M Takahashi, S Ueda, H Misaki, N Sugiyama, K Matsumoto, N Matsuo and S Murao
Asahi Chemical Industry Co., Ltd., Shizuoka-ken, Japan.

We describe a highly sensitive and specific method for determining L- carnitine in serum by use of carnitine dehydrogenase (EC 1.1.1.108). The method involves a new enzymatic cycling technique with NADH, thio- NAD+, and carnitine dehydrogenase, and measures the increase of absorbance at 415 nm of thio-NADH produced at 37 degrees C during the reaction: [formula: see text] The calibration curve for L-carnitine in serum was linear between 5 and 250 mumol/L. Analytical recovery was 96.5-106%, and within-run and between-run imprecisions (CV) were 0.66- 4.33% and 1.02-2.56%, respectively. This method was free from interference by bilirubin, hemoglobin, various acyl-DL-carnitines, and ascorbate. The procedure is simple, rapid, accurate, and automatable. The amount of free L-carnitine in serum (53.6 +/- 11.7 mumol/L, n = 200) was greater in men than in women (45.1 +/- 14.2 mumol/L, n = 200) (mean +/- SD).


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