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Method and apparatus for determining substrate concentration and reagent for determining substrate concentration

a technology of substrate concentration and reagent, which is applied in the direction of liquid/fluent solid measurement, biomass after-treatment, enzymology, etc., can solve the problems of difficult to obtain high determination accuracy and unrealistic approach, and achieve the effect of improving determination accuracy and without any complicated and time-consuming handling steps

Inactive Publication Date: 2010-08-12
ARKRAY INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention relates to a method, apparatus, and reagent for determining the concentration of a substrate, such as glucose, in a specimen without being affected by inhibitors. The invention allows for the measurement of glucose concentration using an enzyme-electrode method while suppressing the reaction between the glucose and the enzyme using a supporting electrolyte. This results in improved accuracy and avoids the need for complicated and time-consuming handling steps. The invention also includes a method for determining the concentration of hydrogen peroxide generated from a substrate using a suppressing agent for reacting with the hydrogen peroxide and an inhibitor.

Problems solved by technology

As a result, the electrode output tends to show a lower value as the reaction between the hemoglobin and the hydrogen peroxide advances in the presence of the hemoglobin, which makes it difficult to obtain high determination accuracy.
It is not avoidable for the specimen to include the hemoglobin by preventing the occurrence of hemolysis of the hemocyte.
This is, however, not a realistic approach because it is necessary to pay a meticulous attention during the transportation, preservation and pretreatment of the blood, which demands burdensome handling steps, and the likelihood of occurrence of hemolyis varies depending on an analyte.

Method used

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  • Method and apparatus for determining substrate concentration and reagent for determining substrate concentration
  • Method and apparatus for determining substrate concentration and reagent for determining substrate concentration
  • Method and apparatus for determining substrate concentration and reagent for determining substrate concentration

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0048]In this example, the influence of sodium azide onto a response value when a glucose concentration in a specimen is determined by means of a hydrogen peroxide electrode was studied.

[0049](Specimen)

[0050]The whole blood in which the glucose concentration is adjusted to 100 mg / dL was used as the specimen.

[0051](Reagent)

[0052]As a reagent, a sample 1 in which NaCl is added by 0.234 wt % to “61H” (manufactured by ARKRAY, Inc.), and a sample 2 in which NaN3 is further added by 0.02 wt % to the sample 1 were respectively diluted 100 times and then supplied into a reaction tank.

[0053](Measurement of Response Value)

[0054]The response value was measured by ADAMS GLUCOSE GA-117 (manufactured by ARKRAY, Inc.) as an oxidation current obtained in a GOD-immobilized hydrogen peroxide electrode when the specimen and reagent were supplied into the reaction tank. The voltage to be applied to the hydrogen peroxide electrode was set to 650 mV, and a response current was measured at 50-msec interva...

example 2

[0058]In this example, the influences of sodium azide and sodium nitrite onto a response value when a glucose concentration in a specimen is determined by means of a hydrogen peroxide electrode were studied.

[0059](Specimen)

[0060]The whole blood in which the glucose concentration is adjusted to 100 mg / dL was used as the specimen.

[0061](Reagent)

[0062]As a reagent, a sample 3 in which NaCl is added by 0.234 wt % to “61H” (manufactured by ARKRAY, Inc.), a sample 4 in which NaN3 is further added by 0.02 wt % to the sample 3, and a sample 5 in which sodium nitrite is added by 0.02 wt % to the sample 3 were respectively diluted 100 times and then supplied into a reaction tank.

[0063](Measurement of Response Value)

[0064]The response value was measured by ADAMS GLUCOSE GA-117 (manufactured by ARKRAY, Inc.) as an oxidation current obtained in a GOD-immobilized hydrogen peroxide electrode when the specimen and reagent were supplied into the reaction tank. The voltage to be applied to the hydrog...

example 3

[0067]In this example, the influence of NaCl in a reagent onto a response value was studied.

[0068]As the reagent, “61H” (manufactured by ARKRAY, Inc.) was prepared as a sample 6, and a sample 7 in which NaCl was added to the sample 6 by 0.234 wt % were used.

[0069]A physiological saline including NaCl of 0.9 wt % was diluted 100 times with the sample 6 or 7, and the obtained solution was used as the specimen.

[0070]The response value was measured in a manner similar to Example 1. The measurement results of response current values, which were converted into voltage values, were illustrated in the drawings, the sample 6 in FIG. 7, while the sample 4 in FIG. 8.

[0071]As is known from FIGS. 7 and 8, the response value was instable in the sample 6 in which NaCl was not added, while the response value was stable in the sample 7 in which NaCl was added. That is, it is considered that NaCl is likely to stabilize the basis of the response current value.

[0072]Therefore, it is considered that the...

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Abstract

The present invention relates to a method, a reagent and an apparatus for determining a substrate concentration based on an amount of hydrogen peroxide generated from a substrate. In the present invention, a suppressing agent for suppressing a reaction between the hydrogen peroxide and an inhibitor is added. As the suppressing agent, an azide compound such as sodium azide or a nitrite compound such as sodium nitrite is used. In the invention, a supporting electrolyte, such as sodium chloride or potassium chloride may be further added.

Description

TECHNICAL FIELD[0001]The present invention relates to a method, an apparatus and a reagent for carrying out the measurement of a substrate such as glucose.BACKGROUND ART[0002]There is a method called an enzyme-electrode method, which is a method for determining a glucose concentration. According to the method, information correlating with a glucose concentration in a specimen is outputted to an electrode in contact with the specimen, and the glucose concentration is calculated based on the output. In an example of the method, electrons are removed at an electrode from hydrogen peroxide generated by oxidizing glucose with an enzyme, and a glucose concentration is calculated based on a current value measured at the time in the electrode (for example, see the Patent Document 1)[0003]When the glucose concentration is determined, blood may be used as the specimen in some cases. In a case where hemolysis in hemocyte occurs, hemoglobin may be included in the specimen whether the whole bloo...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N27/26C01B21/08
CPCC12Q1/28C12Q1/006
Inventor KAMATA, TATSUOTAKAGI, TAKESHI
Owner ARKRAY INC
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