Determination of free and total carnitine in plasma by an enzymatic reaction and spectrophotometric quantitation spectrophotometric determination of carnitine
Carnitine initiates the beta-oxidation of fatty acids and its deficiency is a problem in several metabolic diseases. This work describes a validated quick and simple enzymatic assay for the determination of free and total carnitine in plasma. Carnitine reacts with acetyl-CoA catalized by carnitine a...
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Published in | Clinical biochemistry Vol. 39; no. 10; pp. 1022 - 1027 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
United States
Elsevier Inc
01.10.2006
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Subjects | |
Online Access | Get full text |
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Summary: | Carnitine initiates the beta-oxidation of fatty acids and its deficiency is a problem in several metabolic diseases. This work describes a validated quick and simple enzymatic assay for the determination of free and total carnitine in plasma.
Carnitine reacts with acetyl-CoA catalized by carnitine acetyltransferase. The coenzyme A liberated combines with 5,5′-dithiobis-(2-nitrobenzoic acid) and forms thiophenolate ion, measured spectrophotometically at 412 nm. The method requires precipitation of proteins and the alkaline hydrolysis of acylcarnitines for total carnitine.
The detection limit is 1.7 μmol/L in plasma and inter- and intra-day imprecision is less than 5%. The recovery of spiked plasma samples is 97%. The method was tested with an inter-laboratory assay, yielding excellent correlation (
r
2
=
0.994), and it was applied to the determination of normal values of carnitine in plasma.
A reliable spectrophotometric method has been validated with very good precision, with simple instrumental and easy sample preparation. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0009-9120 1873-2933 |
DOI: | 10.1016/j.clinbiochem.2006.06.005 |