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

[0006]An object of the present invention is to improve determination accuracy by controlling any influences from an inhibitor when the concentration of a substrate, for example, glucose in a specimen such as blood, is determined without any complicated and time-consuming handling steps.Means for Solving the Problems
[0043]Therefore, the reaction of the hydrogen peroxide with the deoxyhemoglobin is suppressed when the suppressing agent is allowed to coexist in the specimen. As a result, the current value measured by the current value measuring unit 32 can be prevented from decreasing, and the variability of the measured response current value can be suppressed in the concentration determination apparatus 1. Thus, the reduction of the result of the concentration determination can be suppressed, which leads to the improvement of reproducibility, and the accuracy and reliability in the measurement can be thereby improved.
[0045]Further, ionic strength in the reaction tank 20 can be increased when the supporting electrolyte such as sodium chloride is included in the reagent, and the oxidation current obtained in the current value measuring unit 32 can be thereby stabilized. Thus, the inclusion of the supporting electrolyte can further improve the measurement reproducibility.
[0056]On the other hand, the result of the specimen 2 in which NaN3 was added showed maximum values larger than those of the specimen 1, and a stabilized time course of the response value. More specifically, in the specimen 2 in which NaN3 was added, it is considered that hemoglobin and NaN3 react with each other, which is expected to prevent the amount of hydrogen peroxide measured in a hydrogen peroxide from decreasing.
[0072]Therefore, it is considered that the response value is stabilized and the determination accuracy and reproducibility are improved when NaCl is added to the reagent.

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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