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Substrate determining method

A technology of substrates and substances, which can be used in biochemical equipment and methods, determination/inspection of microorganisms, measurement devices, etc., and can solve the problems of complex sensor elements or electrode structures, etc.

Inactive Publication Date: 2005-07-13
PANASONIC CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] Thus, in conventional electrochemical methods using direct current potentials for the determination and quantification of specific substrates present in a sample, it is necessary to complicate the structure of sensor elements or electrodes in order to correct or remove the influence of interfering substances

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Dissolve 0.2mM ferrocenecarboxylic acid and 4μm glucose oxidase as the reagent system to obtain 10ml of pH7 phosphate buffer. Put 10ml of this phosphate buffer into a container made of high-hard glass and use it as an electrolyte, use a platinum disk with a diameter of 3mm as a working electrode, and use a square platinum plate with a diameter of 2cm as a counter electrode to form an electrochemical cell. Glucose aqueous solutions in which ascorbic acid was dissolved were added as test solutions so that the concentrations were 0.5 mM and 20 mM (approximately 400 mg / dl).

[0068] After a certain period of time, an AC potential with an amplitude of 0.01V was applied to the working electrode, and its central potential was +0.1V relative to the counter electrode. The central potential on the working electrode is in the range of -0.1V to +0.1V relative to the oxidation-reduction potential of ferrocene carboxylic acid, and the reaction of ascorbic acid on the working electrod...

Embodiment 2

[0073] In this example, the electrochemical cell used in Example 1 was not used, but the biosensor fabricated in the following steps was used, and the measurement was carried out in the same manner as in Example 1. Produced in this example figure 1 and figure 2 The biosensor of the shown structure.

[0074] figure 1 It is an exploded perspective view of the biosensor used in this example without the reagent system. An electrode pattern mask made of resin was provided on an electrically insulating substrate 1 made of glass, and a working electrode 2 and a counter electrode 3 were formed by gold sputtering. A chrome layer is formed between gold and glass as an adhesive layer to improve the adhesion between the two. The working electrode 2 and the counter electrode 3 are electrically connected to the connector for measurement outside the biosensor through wires 4 and 5 respectively.

[0075] After the reagent system layer containing oxidoreductase and electron carrier is ...

Embodiment 3

[0085] This embodiment adopts the electrolyte solution (glucose concentration 20 mM, ascorbic acid concentration 0.5 mM) and electrochemical cell with the same composition as in embodiment 1. After a certain period of time, apply an AC potential on the working electrode, the potential has a certain independent frequency, its central potential is +0.1V relative to the counter electrode, and the amplitude is 0.01V. After a certain period of time, the impedance Z is measured. Then, a direct current potential of +0.3 V was applied to the working electrode for a certain period of time, and the direct current I' flowing between the working electrode and the counter electrode was measured.

[0086] As in Example 1, a series of electrolyte solutions with different glucose concentrations were used, and the measurement and plotting were performed in the same manner as above. As the electrolytic solution with a lower concentration of glucose is used, similar to the result of Example 1, ...

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PUM

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Abstract

Provides a method for accurately quantifying the substrate contained in the test solution using a measurement system with a simple structure so that interfering substances do not cause measurement errors. In a method for quantifying the above-mentioned substrate in a test solution containing dissolved interfering substances and substrates using an electrode system and a reagent system, (a) in the presence of a reagent system containing an oxidoreductase and an electron carrier, The electrode system of the working electrode and the counter electrode supplies a test solution containing dissolved interfering substances and substrates, (b) applying an AC potential to the above-mentioned working electrode, causing the above-mentioned electron carrier to undergo a redox reaction, (c) using the above-mentioned electrode The system measures the electrical signal generated by the above-mentioned redox reaction, and then (d) quantifies the above-mentioned substrate based on the above-mentioned electrical signal.

Description

technical field [0001] The present invention relates to a quantitative method for electrochemically quantifying a substrate contained in a sample using an oxidoreductase. Background technique [0002] Various methods have been developed to easily determine and quantify the presence of specific substrates in a sample. Among them, in recent years, methods for highly selective measurement and quantification using the substrate-selective catalysis of enzymes have attracted attention, and some methods are used as methods for quantifying specific components in body fluids in the field of clinical examinations, as well as self-examinations by ordinary people. field. [0003] The electrochemical quantitative method for glucose will be described as an example of a method for quantitatively measuring a substrate in a sample. Glucose oxidase (hereinafter referred to as GOx) is an enzyme that selectively catalyzes the oxidation of glucose. In the reaction solution containing GOx and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/00C12Q1/26C12Q1/32
CPCC12Q1/001C12Q1/26C12Q1/32G01N27/3273
Inventor 桑畑进中南贵裕
Owner PANASONIC CORP