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Clinical Chemistry 36: 2072-2076, 1990;
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Clinical Chemistry, Vol 36, 2072-2076, Copyright © 1990 by American Association for Clinical Chemistry

Fluorometric determination of carnitine in serum with immobilized carnitine dehydrogenase and diaphorase

K Matsumoto, Y Yamada, M Takahashi, T Todoroki, K Mizoguchi, H Misaki and H Yuki
School of Pharmaceutical Sciences, Toho University, Chiba, Japan.

A fluorometric flow-injection method for determining carnitine with use of immobilized enzymes carnitine dehydrogenase (EC 1.1.1.108) and diaphorase (EC 1.8.1.4) was developed and applied to the assay of carnitine in serum of patients treated with valproic acid. After fractionation and hydrolysis of carnitines in serum samples by perchloric acid and potassium hydroxide, liberated carnitine was converted to resorufin by immobilized carnitine dehydrogenase and diaphorase in the presence of beta-NAD+ (1.0 mmol/L), resazurin (12.5 mumol/L), and Tris acetate (0.6 mol/L, pH 9.0) at 37 degrees C. The fluorescence intensity of resorufin was monitored at lambda Ex 560 nm and lambda Em 580 nm. The calibration curve was linear for carnitine amounts from 0.1 to 1.0 nmol. Quantitative analytical recovery and satisfactory within- and between-run imprecision of carnitine in each carnitine fraction were obtained. Interference by bilirubin, serum albumin, and hemoglobin was negligible. Carnitine deficiencies were detected in about 20% of the valproic acid-treated patients (n = 198). The present method should be useful for monitoring carnitine deficiencies in clinical laboratories.


The following articles in journals at HighWire Press have cited this article:


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 J. Am. Coll. Nutr.Home page
N. Takiyama and K. Matsumoto
Age- and Sex-Related Differences of Serum Carnitine in a Japanese Population
J. Am. Coll. Nutr., February 1, 1998; 17(1): 71 - 74.
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