Enzymatic cycling assay for D-carnitine determination.
Anal Biochem
; 274(1): 34-9, 1999 Oct 01.
Article
in En
| MEDLINE
| ID: mdl-10527494
ABSTRACT
An enzymatic method for d-carnitine determination using the enzyme d-carnitine dehydrogenase is described. The assay is based on the amplified signal produced during NAD(+) cycling in the presence of a tetrazolium salt and using phenazine methosulfate as electron carrier. Optimum assay conditions were studied with two tetrazolium salt pairs 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT)/MTT-formazan and 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl tetrazolium chloride (INT)/INT-formazan. The first pair (MTT) showed higher sensitivity. The calibration curve was linear from 0.1 to 5 mM d-carnitine, with a quantification limit of 0.1 mM and a relative standard deviation of 1.51%. The procedure is simple, rapid, accurate, and easily automated. It was satisfactorily applied to following d-carnitine levels during the microbial transformation of d-carnitine into l-carnitine and to determining the d-carnitine content of pharmaceutical preparations.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Carnitine
/
Alcohol Oxidoreductases
Type of study:
Diagnostic_studies
Language:
En
Journal:
Anal Biochem
Year:
1999
Document type:
Article
Affiliation country: