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1. (WO1980000259) SEMI-QUANTITATIVE ASSAY OF METABOLIC ACIDS
Latest bibliographic data on file with the International Bureau   

Pub. No.:    WO/1980/000259    International Application No.:    PCT/US1979/000498
Publication Date: 21.02.1980 International Filing Date: 16.07.1979
IPC:
C12Q 1/32 (2006.01)
Applicants:
Inventors:
Priority Data:
925946 19.07.1978 US
Title (EN) SEMI-QUANTITATIVE ASSAY OF METABOLIC ACIDS
(FR) DETERMINATION SEMI-QUANTITATIVE D"ACIDES METABOLIQUES
Abstract: front page image
(EN)There is provided a procedure for determining the concentration of a metabolic acid in a biological fluid. The method involves use of a tetrazolium salt, a pyrdine nucleotide, an electron carrier, and an enzyme that is a dehydrogenase for the specific acid being assayed. These components are used to form an assay mixture; thereafter a quantity of the fluid to be assayed is combined with the assay mixture, so that there may commence a reaction in which the tetrazolium salt is changed to a formazan in an amount that is indicative of the concentration of the specific acid. In a preferred embodiment, the procedure permits assay of the concentration of betahydroxybutyrate, and the assay mixture includes components in the relative proportions of .8 micromoles 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride (INT), .32 micromoles phenazine methosulphate (PMS), 1.5 micromoles nicotinamide adenine dinucleotide (NAD), and .75 International Units betahydroxybutyrate dehydrogenase, in a water solution buffered at a pH of approximately 8.5 with hydrogen phosphates of potassium together with glycine and sodium hydroxide and also containing alkylphenoxypolye thyoxyethanol. In another preferred embodiment, the procedure permits assay of lactic acid by means of a similar assay mixture utilizing 27.5 International Units of lactic dehydrogenase in place of the betahydroxybutyrate dehydrogenase used in the previously discussed embodiment. In the latter procedure a preferred embodiment of the assay mixture is buffered at a pH of approximately 9.6 in a glycine-sodium hydroxide buffer and also contains phenoxypolyethyoxyethanol.
(FR)Procede de determination de la concentration d"un acide metabolique dans un fluide biologique. Le procede comprend l"utilisation d"un sel de tetrazolium, une pyridine nucleotide, un porteur d"electrons, et une enzyme qui est une dehydrogenase pour l"acide particulier teste. Ces composants sont utilises pour former un melange d"essai; une quantite du fluide a essayer est combinee au melange d"essai, de maniere a initier une reaction dans laquelle le sel de tetrazolium se transforme en un formazan en une quantite indiquant la concentration de l"acide specifique. Dans un mode preferentiel de realisation, le procede permet de verifier la concentration du betahydroxybutyrate, et le melange d"essai comprend les composants dans les proportions relatives de 0,8 micromole 2-(p-iodophenyle)-3-(p-nitro phenyl)-5-chlorure de phenyltetrazolium (INT), 0,32 micromole de methosulsate de phenasine (PMS), 1,5 micromoles de dinucleotide de nicotilanide adenine (MAD), et 0,75 Unite Internationale de dehydrogenase de betahydroxybutyrate, tamponne a un pH de 8,5 environ avec des phosphates de potassium hydrogenes avec de la glycine et de l"hydroxyde de sodium et contenant egalement de l"alkylphenoxypolyethyoxyethanol. Dans un autre mode preferentiel de realisation, le procede permet la determination de l"acide lactique au moyen d"un melange de determination similaire utilisant 27,5 Unites Internationales de dehydrogenase lactique au lieu de la dehydrogenase betahydroxybutyrate utilisee dans la realisation decrite anterieurement. Dans ce dernier procede un mode preferentiel de realisation du melange de determination est tamponne a un pH de 9,6 environ dans un tampon de glycine-hydroxyde de sodium et contient egalement du phenoxypolyethyoxyethanol.
Designated States:
Publication Language: English (EN)
Filing Language: English (EN)