Dry multilayer analytical element for analysis of enzymes or triglycerides
First Claim
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1. A dry multilayer analytical element for analysis of an enzyme or triglycerides in a fluid sample comprising:
- a support which is impervious to liquids and is light transmissive,a first layer on said support, said first layer comprising a dye forming precursor which is a tetrazolium salt selected from the group consisting of 3,3'"'"'-(3,3'"'"'-dimethoxy-4,4'"'"'-biphenylene)-bis [2-(p-nitrophenyl)-5-phenyltetrazolium chloride] (NBT), 3,3'"'"'-(3,3'"'"'-dimethoxy-4, 4'"'"'-biphenylene)-bis[2,5-diphenyltetrazolium chloride] (BT), 3-(4'"'"',5'"'"'-dimethyltriazolyl-2)-2,4-diphenyltetrazolium bromide (MTT), 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl-tetrazolium chloride (INT), 2,2'"'"',5,5'"'"'-tetra-(p-nitrophenyl)-3,3'"'"'-(3-dimethoxy-4-diphenylene)-ditetrazolium chloride (TNBT), 2,3,5-triphenyltetrazolium chloride (TT) and 3,3'"'"'-(4,4'"'"'-biphenylene)-bis [2,5-diphenyltetrazolium chloride] (NT), anda second layer provided adjacent to said first layer, said second layer comprising an electron-transmitting agent selected from the group consisting of N-methylphenazine methosulfates, meldra Blue, Methylene Blue and diaphorase,wherein;
one of said first layer and said second layer comprises an oxidation co-enzyme, and one of said first layer and said second layer comprises a reagent comprising an enzyme substrate, enzyme or coenzyme other than an oxidation coenzyme, and said oxidation co-enzyme is oxidized nicotinamide adenine dinucleotide (NAD+) or oxidized nicotinamide adenine dinucleotide phosphoric acid (NADP+); and
said reagent is operable to react with said oxidation co-enzyme to form a reduction co-enzyme, in the presence of said enzyme or said triglycerides in said fluid sample, whereby said reduction co-enzyme reduces said electron-transmitting agent to produce a reduced electron-transmitting agent and said reduced electron-transmitting agent reduces said tetrazolium salt to form a dye which can be detected and measured.
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Abstract
A stable dry multi-layer analytical element for enzyme or triglyceride analysis in a fluid sample is prepared containing first and second layers on a support. The first layer contains a tetrazolium salt as a dye forming precursor. The second layer is adjacent the first layer and contains an electron-transmitting agent. Either layer contains a co-enzyme in oxidized form and either layer contains a reagent containing an enzyme substrate, enzyme or co-enzyme other than the oxidized co-enzyme. The analytical element has improved storage stability and low fog density before and after storage.
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Citations
28 Claims
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1. A dry multilayer analytical element for analysis of an enzyme or triglycerides in a fluid sample comprising:
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a support which is impervious to liquids and is light transmissive, a first layer on said support, said first layer comprising a dye forming precursor which is a tetrazolium salt selected from the group consisting of 3,3'"'"'-(3,3'"'"'-dimethoxy-4,4'"'"'-biphenylene)-bis [2-(p-nitrophenyl)-5-phenyltetrazolium chloride] (NBT), 3,3'"'"'-(3,3'"'"'-dimethoxy-4, 4'"'"'-biphenylene)-bis[2,5-diphenyltetrazolium chloride] (BT), 3-(4'"'"',5'"'"'-dimethyltriazolyl-2)-2,4-diphenyltetrazolium bromide (MTT), 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl-tetrazolium chloride (INT), 2,2'"'"',5,5'"'"'-tetra-(p-nitrophenyl)-3,3'"'"'-(3-dimethoxy-4-diphenylene)-ditetrazolium chloride (TNBT), 2,3,5-triphenyltetrazolium chloride (TT) and 3,3'"'"'-(4,4'"'"'-biphenylene)-bis [2,5-diphenyltetrazolium chloride] (NT), and a second layer provided adjacent to said first layer, said second layer comprising an electron-transmitting agent selected from the group consisting of N-methylphenazine methosulfates, meldra Blue, Methylene Blue and diaphorase, wherein; one of said first layer and said second layer comprises an oxidation co-enzyme, and one of said first layer and said second layer comprises a reagent comprising an enzyme substrate, enzyme or coenzyme other than an oxidation coenzyme, and said oxidation co-enzyme is oxidized nicotinamide adenine dinucleotide (NAD+) or oxidized nicotinamide adenine dinucleotide phosphoric acid (NADP+); and said reagent is operable to react with said oxidation co-enzyme to form a reduction co-enzyme, in the presence of said enzyme or said triglycerides in said fluid sample, whereby said reduction co-enzyme reduces said electron-transmitting agent to produce a reduced electron-transmitting agent and said reduced electron-transmitting agent reduces said tetrazolium salt to form a dye which can be detected and measured. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 14, 15, 16, 23, 24, 25, 27)
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- 11. The analytical element according to claim 11 wherein said enzyme is glutamate-oxaloacetate transaminase and said reagent is selected from the group consisting of and said reagent is selected from the group consisting of aspartic acid, alpha-ketoglutaric acid and glutamate dehydrogenase.
Specification