Concentration measuring method, concentration test instrument, and concentration measuring apparatus
First Claim
1. A method for measuring a concentration of a test target, the method comprising:
- constructing a reaction system by supplying the test target to a reagent layer containing an oxidation-reduction enzyme and an electron mediator;
measuring the concentration of the test target by utilizing;
wherein the test target is glucose, the oxidation-reduction enzyme is CyGDH, and a Ru compound is used as the electron mediator,the method further comprising;
applying constant voltage to the reagent layer; and
computing the concentration of the test target on the basis of a response current value from the reagent layer;
wherein the constant voltage is no greater than 300 mV and selected from a range from a standard oxidation-reduction potential (versus a standard hydrogen electrode) between an oxidant and a reductant of the Ru compound through a standard oxidation-reduction potential (versus a standard hydrogen electrode) between ferrocyanide ions and ferricyanide ions.
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Abstract
The present invention relates to technology for constructing a reaction system including a test target, an oxidation-reduction enzyme, and an electron mediator, and measuring the concentration of the test target by an electrochemical process. A Ru compound is used as the electron mediator. The present invention provides a concentration test instrument including a substrate, first and second electrodes formed on the substrate, and a reagent layer formed as a solid. The reagent layer contains an oxidation-reduction enzyme and a Ru compound, and is constituted so as to dissolve and construct a liquid phase reaction system when a sample liquid is supplied.
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Citations
29 Claims
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1. A method for measuring a concentration of a test target, the method comprising:
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constructing a reaction system by supplying the test target to a reagent layer containing an oxidation-reduction enzyme and an electron mediator;
measuring the concentration of the test target by utilizing;wherein the test target is glucose, the oxidation-reduction enzyme is CyGDH, and a Ru compound is used as the electron mediator, the method further comprising; applying constant voltage to the reagent layer; and computing the concentration of the test target on the basis of a response current value from the reagent layer; wherein the constant voltage is no greater than 300 mV and selected from a range from a standard oxidation-reduction potential (versus a standard hydrogen electrode) between an oxidant and a reductant of the Ru compound through a standard oxidation-reduction potential (versus a standard hydrogen electrode) between ferrocyanide ions and ferricyanide ions. - View Dependent Claims (2)
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3. A method for measuring a concentration of a test target, the method comprising:
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contacting whole blood comprising the test target directly with an oxidation-reduction enzyme and an electron mediator disposed between electrodes disposed on a substrate such that the electron mediator is dissolved; applying voltage to the electrodes; and computing the concentration of the test target by amperometry, wherein the oxidation-reduction enzyme is selected from the group consisting of GOD, GDH, α
GDH, CyGDH, cholesterol dehydrogenase, cholesterol oxidase, lactic acid dehydrogenase, lactic acid oxidase, ascorbic acid dehydrogenase, ascorbic acid oxidase, alcohol dehydrogenase, alcohol oxidase, fructose dehydrogenase, 3-hydroxybutyric acid dehydrogenase, pyruvic acid oxidase, uric acid oxidase (uricase), urease, and dihydrolipoamide dehydrogenase (diaphorase), andthe electron mediator is an oxidative ruthenium(III) complex where the complex only contains monodentate ligands. - View Dependent Claims (4, 5, 6, 7, 8, 9, 17, 18)
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10. A concentration measuring apparatus and a measurement test instrument,
wherein the measurement test instrument comprises an oxidation-reduction enzyme; -
an electron mediator; a first electrode; and a second electrode, wherein the oxidation-reduction enzyme is selected from the group consisting of GOD, GDH, α
GDH, CyGDH, cholesterol dehydrogenase, cholesterol oxidase, lactic acid dehydrogenase, lactic acid oxidase, ascorbic acid dehydrogenase, ascorbic acid oxidase, alcohol dehydrogenase, alcohol oxidase, fructose dehydrogenase, 3-hydroxybutyric acid dehydrogenase, pyruvic acid oxidase, uric acid oxidase (uricase), urease, and dihydrolipoamide dehydrogenase (diaphorase);the electron mediator is an oxidative ruthenium(III) complex where the complex only contains monodentate ligands, and the concentration measuring apparatus comprises a voltage applier configured to apply voltage between the first and second electrodes; a current value measurer configured to measure the response current value when voltage is applied between the first and second electrodes; a computer configured to compute the concentration of a test target on the basis of the response current value; and a controller configured to control the current value measurement performed by the current value measurer, wherein the controller is further configured such that the response current value resulting from direct contact of the whole blood with the oxidation-reduction enzyme and the electron mediator and necessary for computation by the computer is measured by the current value measurer. - View Dependent Claims (11, 12, 13, 14, 15, 16, 29)
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19. A device comprising a concentration measuring apparatus together with a concentration test instrument,
wherein: -
the concentration test instrument comprises a substrate;
at least first and second electrodes formed on the substrate;
a solid reagent layer comprising an oxidation-reduction enzyme selected from the group consisting of GOD, GDH, α
GDH and CyGDH; and
an oxidative ruthenium(III) complex where the complex only contains monodentate ligands, wherein the reagent layer is constituted so as to dissolve to a liquid phase reaction system when a sample liquid containing a test target is supplied; andwherein; the concentration measuring apparatus comprises; a voltage applier for applying voltage between the first and second electrodes to oxidize a reductive ruthenium(II) complex to the oxidative ruthenium(III) complex; a current value measurer for measuring the response current value correlated with a quantity of electrons released by the ruthenium(II) complex when voltage is applied between the first and second electrodes; and a computer for computing the concentration of the test target on the basis of the response current value. - View Dependent Claims (20, 21, 22, 23)
where X is selected from the group consisting of NH3, a halogen ion, CN, pyridine, nicotinamide and H2O, and where X determines the valence n+ of the oxidative ruthenium(III) complex.
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24. A method for measuring in blood a concentration of a test target, the method comprising:
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constructing a reaction system containing the test target, an electron mediator and an oxidation-reduction enzyme selected from the group consisting of GOD, GDH, α
GDH and CyGDH;measuring the concentration of the test target by amperometry, wherein an oxidative ruthenium(III) complex only containing monodentate ligands is used as the electron mediator, the method further comprising; a first step of preparing a reductive ruthenium(II) complex in the reaction system; a second step of applying voltage to the reaction system to oxidize the ruthenium(II) complex to the ruthenium(III) complex, and measuring the response current value corresponding to the quantity of electrons released by the ruthenium(II) complex; and a third step of computing the concentration of the test target on the basis of the response current measured in the second step. - View Dependent Claims (25, 26, 27, 28)
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Specification