JP2001116756A - Liquid reagent and preservation method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To solve a problem caused by the use of a refrigerating drying reagent or the like when glucose should be erased when the quantitative examination of a determination substance in liquid is to be made. SOLUTION: The present invention is related to a liquid glucose extinction reagent that sets pH to 6-8.5, and is stable for a long time. Furthermore, a liquid reagent for measuring 1,5-anhydroglucitol using the reagent, and a liquid kit for measuring 1,5-anhydroglucitol measurement are provided.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a liquid glucose elimination reagent, a method for storing the same, a liquid reagent for determining 1,5-anhydroglucitol, and a 1,5-anhydroglucitol (hereinafter, referred to as 1,1) containing them. 5AG) Liquid reagent for measurement,
And a liquid kit for measuring 1,5AG.

[0002]

2. Description of the Related Art Conventionally, when a test substance such as serum or the like in a test liquid is quantitatively tested, glucose must be measured after erasing a large amount of glucose present in the test liquid. , 5AG, maltose, α-glucosidase, glucose amylase or other blood sugars,
There are measurements such as saccharide hydrolases. In order to measure these efficiently in a short time, a glucose elimination reaction is important, and various methods have been proposed. However, it is difficult to ensure the long-term storage stability of aqueous solutions of various enzymes, substrates, and reagents used in the reagents.For example, the measurement reagent is freeze-dried and separated from the reconstitution solution, and the enzyme or substrate is deactivated. A long-term storage stability is ensured by using a reagent composition that avoids the combinations to be used. On the other hand, methods for measuring 1,5AG include (1) a method in which endogenous glucose is adsorbed by an ion exchange resin, and (2) a method in which endogenous glucose is phosphorylated with hexokinase or glucokinase to eliminate it. A 1,5AG measurement kit has been developed and marketed by a method in which hydrogen peroxide generated by oxidizing 5, AG with pyranose oxidase (hereinafter abbreviated as PROD) is detected with a leuco-based chromogenic substrate.

[0003]

However, the method of (1) used for measuring 1,5AG is complicated in operation, and the method of (2) is required to guarantee the stability of the reagent. In addition, it was composed of a lyophilized reagent and a reconstituted solution thereof, and was inconvenient. Therefore, a liquid reagent for measuring 1,5AG which is stable for a long time and requires no preparation at the time of use is desired.

[0004]

Means for Solving the Problems The present inventors have developed a first reagent for eliminating glucose contained in a test solution in a large amount, and 1,
As a result of examining the composition of each enzyme and the reagent and the combination of pH and chromogenic substrate of the second reagent for detecting a measurement test item whose measurement value is affected by glucose such as 5AG, a long-term stable liquid glucose erasing reagent was obtained. And a reagent for a measurement test item whose measurement value is affected by glucose, for example, a 1,5AG measurement reagent, and a kit thereof have been completed. That is, the present invention

(1) A liquid glucose-eliminating reagent containing glucokinase or hexokinase stored at pH 6 to 8.5 (2) Glucokinase or hexokinase stored at pH 6 to 8.5, 4-aminoantipyrine or aniline-based coloring A substrate and a liquid glucose-eliminating reagent containing adenosine triphosphate (3) glucokinase or hexokinase stored at pH 6 to 8.5, 4-aminoantipyrine or aniline-based chromogenic substrate, and adenosine triphosphate, pyruvate kinase; Liquid glucose scavenging reagent containing phosphoenolpyruvate

(4) The method according to any one of (1) to (3), wherein the glucose elimination reagent is contained.
Liquid reagent for measuring 1,5-anhydroglucitol which quantifies by oxidative coloring of 5-anhydroglucitol or its derivative (5) Pyranose oxidase is used to oxidize 1,5-anhydroglucitol or its derivative. Alternatively, the method described in (4) above, wherein L-sorbose oxidase is used,
Liquid reagent for measuring anhydroglucitol (6) Storage method of liquid glucose-eliminating reagent containing glucokinase or hexokinase characterized by adjusting to pH 6 to 8.5 (7) Adjusting to pH 6 to 8.5 A method for storing a liquid glucose-eliminating reagent containing glucokinase or hexokinase, 4-aminoantipyrine or aniline-based chromogenic substrate, and adenosine triphosphate (8) adjusting the pH to 6 to 8.5. For preserving liquid glucose scavenging reagent containing glucokinase or hexokinase, 4-aminoantipyrine or aniline chromogenic substrate, and adenosine triphosphate, pyruvate kinase, phosphoenolpyruvate

(9) A liquid glucose-eliminating reagent containing glucokinase or hexokinase and a liquid reagent for quantifying 1,5-anhydroglucitol which oxidizes and quantifies 1,5-anhydroglucitol or a derivative thereof. (1) A liquid glucose-eliminating reagent containing glucokinase or hexokinase, and 4-aminoantipyrine or aniline-based chromogenic substrate, and 1,5-anhydroglucose 1,5-quantifying by oxidizing sitol or its derivative
(1) Kit for measuring 1,5-anhydroglucitol having good stability comprising a liquid reagent for quantifying anhydroglucitol (11) The above (9) or (9) wherein the pH of the liquid glucose-eliminating reagent is 6 to 8.5. The kit for measuring 1,5-anhydroglucitol described in 10) (12) The pH of the liquid glucose elimination reagent is 6 to 7, and the pH of the liquid reagent for quantifying 1,5-anhydroglucitol is 7 The kit for measuring 1,5-anhydroglucitol according to the above (9) or (10), which is 8.5 to 8.5. (13) Pyranose oxidase or (9) to (12), which are performed using L-sorbose oxidase
A kit for measuring 1,5-anhydroglucitol according to any one of the above.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.
In the present invention, glucokinase or hexokinase and, if necessary, 4-aminoantipyrine (hereinafter referred to as 4A
A liquid glucose-eliminating reagent containing an aniline-based chromogenic substrate, adenosine triphosphate (hereinafter abbreviated as ATP), and optionally pyruvate kinase and phosphoenolpyruvate is pH 6 to pH 8.5, preferably pH 6 to pH 8.5. The pH is adjusted to 6 to 8, more preferably to 6 to 7. In particular, protein and glucose are reversibly bound in stored serum, but by making the protein acidic, glucose is released from the protein, and complete elimination of glucose becomes possible. That is, in this case, the pH is desirably less than 7 in order to accurately determine 1,5AG. Using this liquid glucose elimination reagent, for example, 1,5
When quantifying AG, the pH is preferably 6 to 8.5, more preferably pH 7 containing PROD or L-sorbose oxidase, and peroxidase.
It is used in combination with a liquid reagent (second reagent) for quantifying 1,5-anhydroglucitol which is 8.5. The pH of the solution is preferably around neutral from the viewpoint of the stability and activity of the enzyme. However, since the liquid glucose elimination reagent is more preferably acidic as described above, the second reagent is preferably weakly alkaline. The second reagent may further contain 4AAP or an aniline-based chromogenic substrate. As described above, both the liquid glucose-eliminating reagent (first reagent) and the liquid reagent for determining 1,5-anhydroglucitol (second reagent) can be made liquid. The liquid glucose elimination reagent and the liquid reagent for quantifying 1,5-anhydroglucitol using the enzyme of the present invention can be stably stored at a temperature of 15 ° C. or lower at a temperature at which the reagent does not freeze for a long period of time.

When an aniline-based chromogenic substrate is used, the activity of the coexisting enzyme is less reduced than in the phenol-based chromogenic substrate. As the aniline-based coloring substrate, an aniline derivative having increased water solubility by introducing a sulfoalkyl group or the like called a Neutrinder reagent can be used. For example, N-sulfopropylaniline, N-sulfopropyl-3,5-dimethoxyaniline, N- (2-hydroxy-3-sulfopropyl) -3,5-dimethoxyaniline, N-ethyl-N-sulfur Phopropylaniline, N-ethyl-N- (2-hydroxy-3-sulfopropyl) aniline, N-ethyl-N-sulfopropylanisidine, N-ethyl-N-sulfopropyl-m
-Toluidine, N-ethyl-N- (2-hydroxy-3
-Sulfopropyl) aniline, N-ethyl-N- (2
-Hydroxy-3-sulfopropyl) -m-anisidine, N-ethyl-N- (2-hydroxy-3-sulfopropyl) -m-toluidine (hereinafter abbreviated as TOOS),
N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3,5-dimethoxyaniline, N, N-bissulfopropylaniline, N, N-bissulfopropyl-m-toluidine, N, N-bis (2-hydroxy-
3-sulfopropyl) aniline, N, N-bis (2-
(Hydroxy-3-sulfopropyl) -m-toluidine, N-ethyl-N- (2-succinylaminoethyl)
-M-toluidine and the like.

An aniline type and a m-toluidine type having a large molar extinction coefficient are preferred, and TOOS, N, N-bis (2-hydroxy-3-sulfopropyl) aniline,
N, N-bissulfopropylaniline, N, N-bissulfopropyl-m-toluidine, N, N-bis (2
-Hydroxy-3-sulfopropyl) -m-toluidine, N-ethyl-N- (2-succinylaminoethyl)
-M-toluidine and the like. Various test liquids can be used, and examples thereof include cerebrospinal fluid, plasma, serum, urine and other body fluids and tissue extracts of various animals including humans, plant extracts, foods and the like.

The 1,5AG derivative used in the present invention is not particularly limited as long as it can measure 1,5AG by oxidative coloration. For example, 1,5AG phosphate ester and 1,5AG organic carboxylic acid Esters and the like, and in particular, 1,5AG-6-phosphoric acid, 1,5AG-6-
Acetate is preferred. The determination of 1,5-anhydroglucitol or its derivative by oxidative coloring in the present invention means that the substance is oxidized by an enzyme using 1,5-anhydroglucitol or its derivative as a substrate and is converted or its conversion. The objective is to determine the amount of 1,5AG by color development of the substrate.

The glucokinase and hexokinase used in the present invention are glucokinase: EC 2.7.1.2 and hexokinase by IUPAC-IUB nomenclature.
There is no particular limitation as long as it can be classified into EC 2.7.1.1, and a commercially available product can be used. The oxidizing enzyme used for oxidative coloring of 1,5AG or a derivative thereof used in the present invention is not particularly limited. For example, pyranose oxidase, L-sorbose oxidase,
5-anhydroglucitol dehydrogenase, 1,5-anhydroglucitol 6-phosphate dehydrogenase and the like,
Pyranose oxidase and L-sorbose oxidase include pyranose oxidase: EC 1.1.3.1.1.
0, L-sorbose oxidase: EC 1.1.3.1.
There is no particular limitation as long as it can be classified into 1, and a commercially available product can be used. The pyruvate kinase and peroxidase used in the present invention include pyruvate kinase: EC 2.7.1.40 and peroxidase: EC
There is no particular limitation as long as it can be classified into 1.1.1.1.7, and a commercially available product can be used.

The amounts of glucokinase and hexokinase used vary depending on the amount of glucose in the test solution and the like, but are usually as follows. Glucokinase (hereinafter abbreviated as GK): 0.1 U / mL to 500 U / mL, preferably 1 U / mL to 20 U / mL Hexokinase: 0.1 U / mL to 500 U / mL, preferably 1 U / mL to 20 U / mL The amount of other enzymes and substrates used varies depending on the measurement target and the like, but is usually as follows. Pyruvate kinase (hereinafter abbreviated as PK): 1 U / mL to 10 U / mL PROD: 5 U / mL to 100 U / mL L-sorbose oxidase: 5 U / mL to 100 U / mL Peroxidase (hereinafter abbreviated as POD): 0.5 U / mL to 20 U / mL ATP: 0.1 mmol / L to 3 mmol / L phosphoenolpyruvate (hereinafter abbreviated as PEP): 2 mmol / L to 12 mmol / L 4AAP: 0.1 mmol / L to 10 mmol / L aniline-based chromogenic substrate : 0.2 mmol / L to 10 mmol / L

The measurement using the measuring reagent of the present invention is usually performed
It is performed in a buffer of pH 6.0 to 8.5 and 10 to 200 mM at 10 to 55 ° C. for about 1 to 50 minutes. The buffer is preferably a Tris-HCl buffer, a Good's buffer, or the like.
The concentration is preferably about 00 mM and about 0.5 to 50 mM of magnesium chloride, but is not particularly limited.

The stable 1,5AG measuring kit of the present invention comprises a liquid glucose elimination reagent containing GK or hexokinase and a 1,5AG for oxidizing and quantifying 1,5-anhydroglucitol or a derivative thereof. -It comprises at least two liquid agents of a liquid reagent for quantifying anhydroglucitol.
These solutions have good storage stability and can be stably stored at 10 ° C. or lower for one year or more. The liquid glucose elimination reagent is
The liquid glucose elimination reagent containing GK or hexokinase described above can be used. Specifically, GK or hexokinase, if necessary, further 4AAP or aniline-based chromogenic substrate, and ATP, and if necessary, PK, P
Including EP, pH 6 to pH 8.5, preferably pH 6 to pH 8.5
8, more preferably pH 6-7. As the liquid reagent for quantifying 1,5-anhydroglucitol, a liquid reagent for quantifying 1,5-anhydroglucitol which oxidizes and quantifies the aforementioned 1,5-anhydroglucitol or a derivative thereof is used. And specifically includes PROD or L-sorbose oxidase, and POD, and has a pH of preferably 6 to 8.5, more preferably pH 7 to 8.5. The enzymes and reagents in this kit are used at the above concentrations. In addition, this kit is applicable to various automatic analyzers, and can be used for 1,5AG measurement, for example, using the parameters described in Example 1 below. The same sample as described above can be used as a sample that can use this kit.

[0016]

EXAMPLES Hereinafter, the present invention will be described specifically with reference to Comparative Examples and Examples, but the present invention is not limited to these Examples.

Example 1 A liquid glucose-eliminating reagent according to the present invention was prepared, and the result of measuring a 1000 mg / dL aqueous solution of glucose as a test solution is shown in Table 1. The measurement was performed using an automatic analyzer 7150 (manufactured by Hitachi, Ltd.) with the following parameters. The amount of residual glucose was physiological saline and glucose 50 mg / d.
It was determined from the calibration curve of L. Analysis method 2-point end Measurement point 27-50 Sample volume 6 μL First reagent 180 μL Second reagent 90 μL Temperature 37 ° C. Measurement wavelength (secondary / positive) 700/546 nm Reagent composition First reagent HEPES buffer (pH 7.5) 100 mM potassium chloride 50 mM magnesium chloride 5 mM TOOS 3.0 mM GK 4 U / mL PK 4 U / mL ATP 1.0 mM PEP 8.0 mM NaN ₃ 0.02% Second reagent HEPES buffer (pH 7.5) 100 mM NaN ₃ 0.02% horseradish Peroxidase (hereinafter abbreviated as HRP) 5 U / mL Glucose oxidase 40 U / mL 4 AAP 1.5 mM

Table 1 Measurement value of glucose 1000 mg / dL (mg / dL) Reagent storage period (month) Storage temperature 0 13 7 13 10 ° C. 0.0 0.0 0.0 0.0 0.0 15 ° C. 0 0.0 0.0 0.0 0.0 0.0 25 ° C. 0.0 0.0 0.0 0.0 3.0
It was stable for 1 year or more at below ℃.

Example 2 A liquid glucose-eliminating reagent according to the present invention was prepared, and the results were obtained by preparing a reagent having the following composition and measuring a 1000 mg / dL aqueous solution of glucose as a test solution. The measurement was performed by an automatic analyzer 7150 type (manufactured by Hitachi, Ltd.).
And using the same parameters as in Example 1. The amount of residual glucose was 500 μl of physiological saline and glucose.
It was determined from a calibration curve of g / mL. Reagent composition First reagent MES buffer (pH 6.2) 100 mM potassium chloride 50 mM magnesium chloride 5 mM 4AAP 1.5 mM GK 8 U / mL PK 4 U / mL ATP 1.0 mM PEP 8.0 mM NaN ₃ 0.02% Second reagent HEPES Buffer (pH 7.5) 100 mM NaN ₃ 0.02% HRP 5 U / mL Glucose oxidase 40 U / mL TOOS 6.0 mM

Table 2 Measured value of glucose 1000 mg / dL (mg / dL) Reagent storage period (month) Storage temperature 0 13 7 13 10 ° C. 0.0 0.0 0.0 0.0 0.0 15 ° C. 0 0.0 0.0 0.0 2.7 15.0 25 ° C. 0.0 0.0 22.4 215.6 Unmeasured As described above, it was stable for 1 year at 10 ° C. or less while still in a liquid state.

Example 3 A liquid glucose-eliminating reagent according to the present invention was prepared. A reagent having the following composition was prepared, and a 1000 mg / dL aqueous glucose solution was measured as a test solution. Table 3 shows the results. The measurement was performed by an automatic analyzer 7150 type (manufactured by Hitachi, Ltd.).
And using the same parameters as in Example 1. The amount of residual glucose was 500 μl of physiological saline and glucose.
It was determined from a calibration curve of g / mL. Reagent composition First reagent Tris-HCl buffer (pH 8.4) 100 mM potassium chloride 50 mM magnesium chloride 5 mM 4AAP 1.5 mM GK 8 U / mL PK 4 U / mL ATP 1.0 mM PEP 8.0 mM NaN ₃ 0.02% Second reagent HEPES buffer (pH 7.5) 100 mM NaN ₃ 0.02% HRP 5 U / ml Glucose oxidase 40 U / ml TOOS 6.0 mM

Table 3 Measured value of glucose 1000 mg / dL (mg / dL) Reagent storage period (month) Storage temperature 0 13 7 13 10 ° C. 0.0 0.0 0.0 0.0 0.0 15 ° C. 0 0.0 0.0 0.0 0.3 2.5 25 ° C. 0.0 0.0 0.2 43.7 218.4 As described above, it was stable for one year at 10 ° C. or less while remaining liquid.

Example 4 A liquid glucose-eliminating reagent according to the present invention was prepared. A reagent having the following composition was prepared, and a 1000 mg / dL aqueous solution of glucose was measured as a test solution. The results are shown in Table 4. The measurement was performed by an automatic analyzer 7150 type (manufactured by Hitachi, Ltd.).
And using the same parameters as in Example 1. The amount of residual glucose was 500 μl of physiological saline and glucose.
It was determined from a calibration curve of g / mL. Reagent composition First reagent MES buffer (pH 6.2) 100 mM potassium chloride 50 mM magnesium chloride 5 mM N-ethyl-N- (2-hydroxy-3-sulfopropyl) aniline 6.0 mM GK 8 U / mL PK 4 U / mL ATP 1.0 mM PEP 8.0 mM NaN ₃ 0.02% Second reagent HEPES buffer (pH 7.5) 100 mM NaN ₃ 0.02% HRP 5 U / ml Glucose oxidase 40 U / ml 4AAP 1.5 mM

Table 4 Measured value of glucose 1000 mg / dL (mg / dL) Reagent storage period (month) Storage temperature 0 13 7 13 10 ° C. 0.0 0.0 0.0 0.0 0.0 15 ° C. 0 0.0 0.0 0.0 0.0 0.2 25 ° C. 0.0 0.0 0.0 48.9 201.5 As described above, it was stable for one year at 15 ° C. or less while remaining liquid.

Example 5 1,5AG measuring reagent according to the present invention A reagent having the following composition was prepared, and the 1,5AG concentration was measured using serum as a test solution. Table 5 shows the results. The measurement was performed using an automatic analyzer 7150 (manufactured by Hitachi, Ltd.) with the following parameters, and the measured values of 1,5AG were as follows: 1,5AG concentration 0 μg / mL and 50 μg / mL.
Was determined from the calibration curve. Analysis method 2-point end Measurement point 27-50 Sample volume 6 μL First reagent 180 μL Second reagent 90 μL Temperature 37 ° C. Measurement wavelength (secondary / positive) 700/546 nm In addition, 1,5AG and glucose of the test serum used for measurement The concentrations were as follows: 1,5AG (μg / mL) Glucose (mg / dL) Serum A 4.5 256 Serum B 14.6 111 Serum C 23.8 109 Reagent composition First reagent HEPES buffer (pH 7.5) 100 mM Potassium chloride 50 mM Chloride Magnesium 5 mM 4AAP 1.5 mM GK 4 U / mL PK 4 U / mL ATP 1.0 mM PEP 8.0 mM NaN ₃ 0.02% Second reagent HEPES buffer (pH 7.5) 100 mM NaN ₃ 0.02% HRP 5 U / mL PROD 40U / mL TOOS 6.0mM

Table 5 Measured value of 1,5AG when serum is used as a test liquid (μg / mL) Reagent storage period (months) Storage temperature 0 13 7 13 10 ° C. Serum A 4.5 4.4 4.5 4.7 4.6 Serum B 14.2 14.5 14.6 14.5 14.6 Serum C 24.3 23.8 24.6 24.2 23.9 15 ° C Serum A 4.5 4.2 4.6 4.7 4.8 Serum B 14.2 14.5 14.9 14.7 14.9 Serum C 24.3 24.4 24.5 24.2 24.6 25 ° C Serum A 4.5 4.3 4.2 4.2 5.2 Serum B 14.2 14.4 14.4 14.1 14.9 Serum C 24.3 24.2 24.2 24.3 25.5 As above Stable for a long time in liquid state, especially 15
It was stable for 1 year or more at below ℃.

Comparative Example 1 A reagent having the following composition was prepared, and a 1000 mg / dL aqueous solution of glucose was measured as a test solution. The results are shown in Table 6. The measurement was performed by an automatic analyzer 7150 type (manufactured by Hitachi, Ltd.).
And using the same parameters as in Example 1. The amount of residual glucose was 500 μl of physiological saline and glucose.
It was determined from a calibration curve of g / mL. Reagent composition First reagent MES buffer (pH 5.8) 100 mM potassium chloride 50 mM magnesium chloride 5 mM 4AAP 1.5 mM GK 8 U / mL PK 4 U / mL ATP 1.0 mM PEP 8.0 mM NaN ₃ 0.02% Second reagent HEPES Buffer (pH 7.5) 100 mM NaN ₃ 0.02% HRP 5 U / ml Glucose oxidase 40 U / ml TOOS 6.0 mM

Table 6 Measured value of glucose 1000 mg / dL (mg / dL) Reagent storage period (month) Storage temperature 0 13 7 13 10 ° C. 0.0 0.0 0.0 12.8 95.4 15 ° C. 0 0.0 0.0 18.9 95.2 Calibration curve over 25 ° C 0.0 4.8 264.2 Unmeasured As described above, glucose is 10 after 7 months even at 10 ° C.
The complete erasure ability of 00 mg / dL was lost.

Comparative Example 2 A reagent having the following composition was prepared, and a 1000 mg / dL aqueous solution of glucose was measured as a test solution. The results are shown in Table 7. The measurement was performed by an automatic analyzer 7150 type (manufactured by Hitachi, Ltd.).
And using the same parameters as in Example 1. The amount of residual glucose was 500 μl of physiological saline and glucose.
It was determined from a calibration curve of g / mL. Reagent composition First reagent Tris-HCl buffer (pH 8.8) 100 mM potassium chloride 50 mM magnesium chloride 5 mM 4 AAP 1.5 mM GK 8 U / mL PK 4 U / mL ATP 1.0 mM PEP 8.0 mM NaN ₃ 0.02% Second reagent HEPES buffer (pH 7.5) 100 mM NaN ₃ 0.02% HRP 5 U / ml Glucose oxidase 40 U / ml TOOS 6.0 mM

Table 7 Measurement value of glucose 1000 mg / dL (mg / dL) Reagent storage period (month) Storage temperature 0 13 7 13 10 ° C. 0.0 0.0 0.0 1.9 48.5 15 ° C. 0 0.0 0.0 0.6 9.8 122.2 25 ° C. 0.0 0.0 15.6 158.3 unmeasured As described above, even after 10 months, glucose 10
The complete erasure ability of 00 mg / dL was lost.

[0031]

The liquid glucose elimination reagent of the present invention, which is stable for a long period of time and does not need to be prepared at the time of use, does not require complicated operations when glucose in the sample must be removed, and is convenient to use. Furthermore, it has become possible to provide a liquid reagent for measuring 1,5-anhydroglucitol, which does not require any dissolution, and a kit for measuring 1,5AG, which is composed of the liquid reagent and does not require any dissolution, using this glucose-eliminating reagent.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) G01N 33/48 G01N 33/48 C

Claims (13)

[Claims] 1. A liquid glucose-eliminating reagent containing glucokinase or hexokinase stored at pH 6 to 8.5. 2. A liquid glucose-eliminating reagent containing glucokinase or hexokinase, 4-aminoantipyrine or aniline-based chromogenic substrate stored at pH 6 to 8.5, and adenosine triphosphate. 3. A liquid glucose-eliminating reagent containing glucokinase or hexokinase, 4-aminoantipyrine or aniline-based chromogenic substrate stored at pH 6 to 8.5, and adenosine triphosphate, pyruvate kinase, and phosphoenolpyruvate. 4. A method for quantifying 1,5-anhydroglucitol or a derivative thereof by oxidative coloring, comprising the glucose scavenging reagent according to claim 1. Liquid reagent for measuring glucitol 5. The method according to claim 4, wherein the oxidation of 1,5-anhydroglucitol or a derivative thereof is carried out using pyranose oxidase or L-sorbose oxidase.
Liquid reagent for measuring 5-anhydroglucitol 6. A method for storing a liquid glucose-eliminating reagent containing glucokinase or hexokinase, wherein the pH is adjusted to 6 to 8.5. 7. A method for preserving a liquid glucose-eliminating reagent containing glucokinase or hexokinase, 4-aminoantipyrine or aniline-based chromogenic substrate, and adenosine triphosphate, wherein the pH is adjusted to 6 to 8.5. 8. A composition comprising glucokinase or hexokinase, 4-aminoantipyrine or aniline chromogenic substrate, and adenosine triphosphate, pyruvate kinase, phosphoenolpyruvate, characterized in that the pH is adjusted to 6 to 8.5. Storage method of liquid glucose elimination reagent 9. A liquid glucose elimination reagent containing glucokinase or hexokinase, and 1,5-anhydroglucitol or a derivative thereof, which is oxidized and quantified.
Kit for measuring 1,5-anhydroglucitol with good stability, comprising a liquid reagent for determining anhydroglucitol 10. A glucokinase or hexokinase,
And a liquid glucose elimination reagent containing a 4-aminoantipyrine or aniline-based chromogenic substrate, and a liquid reagent for quantifying 1,5-anhydroglucitol which oxidizes and quantifies 1,5-anhydroglucitol or a derivative thereof. Kit for measuring 1,5-anhydroglucitol with good stability 11. The pH of the liquid glucose elimination reagent is 6 to
The kit for measuring 1,5-anhydroglucitol according to claim 9 or 10, which is 8.5. 12. The pH of the liquid glucose elimination reagent is 6-7.
The kit for measuring 1,5-anhydroglucitol according to claim 9 or 10, wherein the pH of the liquid reagent for determining 1,5-anhydroglucitol is 7 to 8.5. 13. The method of claim 1, wherein the oxidation of 1,5-anhydroglucitol or a derivative thereof is performed by pyranose oxidase or L-
The method is performed using sorbose oxidase.
The kit for measuring 1,5-anhydroglucitol according to any one of the above