Highly sensitive assay method for L-carnitine and composition for practicing same
First Claim
1. A method of assaying L-carnitine comprising reacting a specimen containing L-carnitine with the following reagents:
- a) L-carnitine dehydrogenase having coenzymes of thionicotinamide adenine dinucleotide group (thio-NAD group) and nicotinamide adenine dinucleotide group (NAD group), and which catalyzes a reversible reaction forming dehydrocarnitine from a substrate of L-carnitine;
b) A1 ;
c) B1 ;
to effect a cycling reaction of the formula ##STR5## wherein A1 is thio-NAD group or NAD group, A2 is a reduced form of A1, when A1 is thio-NAD group, B1 is reduced NAD group and when A1 is NAD group, B1 is reduced thio-NAD, and wherein B2 is an oxidized form of B1 ; and
measuring an amount of A2 or B1 generated or consumed by the cycling reaction.
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Abstract
A method of assaying L-carnitine in a specimen comprises reacting a specimen containing L-carnitine with:
a) L-carnitine dehydrogenase having coenzymes of the thio-NAD group and of the NAD group, and which catalyzes a reversible reaction forming dehydrocarnitine from a substrate of carnitine,
b) A1 and
c) B1
to effect a cycling reaction of the formula ##STR1## wherein A1 is thio-NAD group or NAD group, A2 is a reduced form of A1, when A1 is thio-NAD group, B1 is reduced NAD group and when A1 is NAD group, B1 is reduced thio-NAD, and wherein B2 is an oxidized form of B1 ; and measuring an amount of A2 or B1 generated or consumed by the cycling reaction. A composition for performing the assay comprises the above L-carnitine dehydrogenase, as well as the above components A1 and B1.
10 Citations
10 Claims
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1. A method of assaying L-carnitine comprising reacting a specimen containing L-carnitine with the following reagents:
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a) L-carnitine dehydrogenase having coenzymes of thionicotinamide adenine dinucleotide group (thio-NAD group) and nicotinamide adenine dinucleotide group (NAD group), and which catalyzes a reversible reaction forming dehydrocarnitine from a substrate of L-carnitine; b) A1 ; c) B1 ; to effect a cycling reaction of the formula ##STR5## wherein A1 is thio-NAD group or NAD group, A2 is a reduced form of A1, when A1 is thio-NAD group, B1 is reduced NAD group and when A1 is NAD group, B1 is reduced thio-NAD, and wherein B2 is an oxidized form of B1 ; and measuring an amount of A2 or B1 generated or consumed by the cycling reaction. - View Dependent Claims (2, 3, 4, 5)
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6. A composition for assaying L-carnitine, comprising:
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a) L-carnitine dehydrogenase having coenzymes of thionicotinamide adenine dinucleotide group (thio-NAD group) and nicotinamide adenine dinucleotide group (NAD group), and which catalyzes a reversible reaction forming dehydrocarnitine from a substrate of L-carnitine; b) A1 ; c) B1 ; wherein A1 is thio-NAD group or NAD group, when A1 is thio-NAD group, B1 is reduced form of NAD group, and when A1 is NAD group, B1 is reduced form of thio-NAD group. - View Dependent Claims (7, 8, 9, 10)
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Specification
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- Resources
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Current AssigneeAsahi Kasei Kogyo Kabushiki Kaisha
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Original AssigneeAsahi Kasei Kogyo Kabushiki Kaisha
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InventorsTakahashi, Mamoru, Ueda, Shigeru
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Primary Examiner(s)Wityshyn, Michael G.
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Assistant Examiner(s)Mohamed, Abdel A.
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Application NumberUS07/640,117Time in Patent Office1,054 DaysField of Search435/26, 435/89, 435/90, 435/117, 435/128, 435/829, 436/815US Class Current435/26CPC Class CodesC12N 9/0006 acting on CH-OH groups as d...C12Q 1/26 involving oxidoreductaseC12Q 1/32 involving dehydrogenaseY10S 435/829 AlcaligenesY10S 436/815 Test for named compound or ...
