Inhibitor of 2,5-dihydroxybenzenesulfonic acid or its salt, and reagent composition for measurement that suppresses the effect of 2,5-dihydroxybenzenesulfonic acid or its salt on measurement

Abstract

To provide an inhibitor for 2,5-dihydroxybenzenesulfonic acid or salt thereof, and a measurement reagent composition for suppressing the effect of 2,5-dihydroxybenzenesulfonic acid or salt thereof on measurement when measuring substances contained in a sample.SOLUTION: An embodiment of the present invention provides a measurement reagent composition comprising peroxidase, a coloring reagent configured to develop color by reacting with hydrogen peroxide derived from a substance in a sample under the presence of peroxidase, and cyclodextrin or an analog thereof.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to an inhibitor for 2,5-dihydroxybenzenesulfonic acid or a salt thereof, and a reagent composition for measurement that suppresses the influence of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on measurement. [Background technology]

[0002] Measurement systems using Trinder's reagent and couplers, which are redox color-developing systems utilizing the hydrogen peroxide / peroxidase reaction, and measurement systems using leuco-type dyes are used for quantitative evaluation in various clinical biochemical tests (see, for example, Patent Documents 1 and 2). Although the sensitivity of these measurement systems has been increased with improvements in color-developing reagents, it is known that the measured values ​​are affected by substances such as 2,5-dihydroxybenzenesulfonic acid such as ethamsylate, ascorbic acid, N-acetylcysteine, and bilirubin in biological samples. In recent years, it has been suggested that calcium dobesilate, which is a calcium salt of 2,5-dihydroxybenzenesulfonic acid, also affects the measured values ​​in the measurement systems using Trinder's reagent and couplers.

[0003] Calcium dobesilate is a drug with vascular protective properties, and is prescribed as a treatment for diabetic retinopathy, chronic venous insufficiency, and various microvascular disorders. It has also been shown to have protective effects against diabetic nephropathy and acute kidney injury caused by gentamicin, and its protective ability against intestinal ischemia-reperfusion injury due to increased antioxidant capacity has also been suggested. Orally administered calcium dobesilate remains in the body as the parent drug even after absorption, and is excreted from the kidney and intestine. According to pharmacokinetic data, the plasma concentration of calcium dobesilate reaches 8 μg / mL 6 hours after a single dose of 500 mg. The current clinical dosage form is 500 mg three times a day, so the steady-state plasma concentration is estimated to be about 15 μg / mL (see, for example, Non-Patent Document 1).

[0004] X. Guo et al. have demonstrated that calcium dobesilate has a dose-dependent effect on the measured values ​​of uric acid, triglycerides, total cholesterol, good cholesterol, and bad cholesterol in samples containing calcium dobesilate, when the samples were measured using a color-developing system with Trinder's reagent and a coupler. X. Guo et al. have also found that calcium dobesilate has a dose-dependent effect on the measured values ​​of glycated albumin (GA). In other words, by preparing samples with calcium dobesilate added at multiple concentrations to three types of serum with different amounts of glycated albumin and measuring the glycated albumin in the samples, they have demonstrated that calcium dobesilate has an effect on the measured values ​​of glycated albumin by approximately 10% at 16 μg / mL and approximately 30% at 64 μg / mL. Furthermore, by administering 500 mg of calcium dobesilate 3 times a day for 3 days to healthy subjects and then measuring their fasting blood after reaching a steady state, and then measuring their blood two hours after administering a further 500 mg of calcium dobesilate, it was found that calcium dobesilate of 7.33 μg / mL in fasting conditions affected glycated albumin measurements by 4.8%, and calcium dobesilate of 18.52 μg / mL two hours later affected glycated albumin measurements by 12.8% (see, for example, non-patent documents 2 and 3).

[0005] Glycated albumin is one of the glycosylated proteins that is a very important index in diagnosing and managing diabetes. Glycated albumin has been introduced into clinical testing as an item that reflects the average blood glucose level for the past about two weeks, which is more recent than hemoglobin A1c (HbA1c), which reflects the average blood glucose level for the past about one to two months. Glycated albumin is measured using an oxidation-reduction coloring system that utilizes the hydrogen peroxide / peroxidase reaction described above. Measurement systems using oxidation-reduction coloring systems, including glycated albumin, are used to measure various target substances in clinical biochemistry tests, and as described above, it has been found that the measured value is affected by 2,5-dihydroxybenzenesulfonic acid or a salt thereof. On the other hand, no method is known for reducing the effect. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Patent No. 4697809 [Patent Document 2] Patent No. 5274590 [Non-patent literature]

[0007] [Non-Patent Document 1] Xiuzhi Guo et al., “Negative interferences by calcium dobesilate in the detection of five serum analytes Trinder reaction-based assays,” PLoS ONE, 13(2):e0192440, February 2018. [Non-Patent Document 2] X. Guo et al., “Negative interference by Calcium Dobesilate in Five Trinder Reaction Assays,” 69th AACC Annual Scientific Meeting Abstracts, A-244, 2017. [Non-Patent Document 3] X. Guo et al., “Calcium Dobesilate can Negatively Interfere with the Detection of Glycated Albumin”, 69th AACC Annual Scientific Meeting Abstracts, A-245, 2017. Summary of the Invention [Problem to be solved by the invention]

[0008] An object of the present invention is to provide an inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof, and a measurement reagent composition that suppresses the effect of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on the measurement when measuring a substance in a sample. [Means for solving the problem]

[0009] Aspects of the invention include the following.

[0010] [1] A measurement reagent composition comprising peroxidase, a color-developing reagent configured to react with hydrogen peroxide derived from a substance in a sample in the presence of peroxidase to develop color, and cyclodextrin or an analog thereof.

[0011] [2] The reagent composition for measurement according to [1], which contains an effective amount of peroxidase.

[0012] [3] The reagent composition for measurement according to [1] or [2], comprising an effective amount of a color-developing reagent.

[0013] [4] The reagent composition for measurement according to any one of [1] to [3], which contains an effective amount of cyclodextrin or an analogue thereof.

[0014] [5] The reagent composition for measurement according to any one of [1] to [4], wherein the color-developing reagent comprises a Trinder reagent and a coupler.

[0015] [6] Trinder's reagent is N-(3-sulfopropyl)aniline sodium monohydrate (HALPS), N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium monohydrate (TOPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (MAOS), N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (HDAPS), N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (HDAOS), N-ethyl-N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (DAPS), N-ethyl-N-(2-hydroxy- ... The reagent composition for measurement according to [5], which is one selected from the group consisting of N-ethyl-N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium (DAOS), N-ethyl-N-(3-sulfopropyl)aniline sodium (ALPS), N-ethyl-N-(3-sulfopropyl)-3-methoxyaniline sodium monohydrate (ADPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methoxyaniline sodium dihydrate (ADOS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine (TOOS), N,N-bis(4-sulfobutyl)-3-methylaniline disodium (TODB), and 3-hydroxy-2,4,6-triiodobenzoic acid (HTIB).

[0016] [7] The reagent composition for measurement according to [6], wherein the Trinder reagent is TODB.

[0017] [8] The reagent composition for measurement according to any one of [5] to [7], wherein the concentration of the Trinder reagent is 0.1 mmol / L or more and 50 mmol / L or less.

[0018] [9] The reagent composition for measurement according to [8], wherein the concentration of the Trinder reagent is 0.2 mmol / L or more.

[0019]

[10] The reagent composition for measurement according to [8], wherein the concentration of the Trinder reagent is 0.5 mmol / L or more.

[0020]

[11] The reagent composition for measurement according to [8], wherein the concentration of the Trinder reagent is 1.0 mmol / L or more.

[0021]

[12] The reagent composition for measurement according to [8], wherein the concentration of the Trinder reagent is 30 mmol / L or less.

[0022]

[13] The reagent composition for measurement according to [8], wherein the concentration of the Trinder reagent is 20 mmol / L or less.

[0023]

[14] The reagent composition for measurement according to [8], wherein the concentration of the Trinder reagent is 10 mmol / L or less.

[0024]

[15] The reagent composition for measurement according to any one of [8] to

[14] , wherein the concentration of the Trinder reagent is the concentration of the Trinder reagent in the reagent composition for measurement.

[0025]

[16] The reagent composition for measurement according to any one of [8] to

[14] , wherein the concentration of the Trinder reagent is the concentration of the Trinder reagent in a mixture of the reagent composition for measurement and a sample.

[0026]

[17] The reagent composition for measurement according to any one of [5] to

[16] , wherein the coupler is one selected from the group consisting of 4-aminoantipyrine (4-AA), aminoantipyrine derivatives, vanillindiaminesulfonic acid, methylbenzthiazolinone hydrazone (MBTH), and sulfonated methylbenzthiazolinone hydrazone (SMBTH).

[0027]

[18] The reagent composition for measurement according to

[17] , wherein the coupler is 4-AA.

[0028]

[19] The reagent composition for measurement according to any one of [5] to

[18] , wherein the concentration of the coupler is 0.1 mmol / L or more and 50 mmol / L or less.

[0029]

[20] The reagent composition for measurement according to

[19] , wherein the concentration of the coupler is 0.2 mmol / L or more.

[0030]

[21] The reagent composition for measurement according to

[19] , wherein the concentration of the coupler is 0.5 mmol / L or more.

[0031]

[22] The reagent composition for measurement according to

[19] , wherein the concentration of the coupler is 1.0 mmol / L or more.

[0032]

[23] The reagent composition for measurement according to

[19] , wherein the concentration of the coupler is 30 mmol / L or less.

[0033]

[24] The reagent composition for measurement according to

[19] , wherein the concentration of the coupler is 20 mmol / L or less.

[0034]

[25] The reagent composition for measurement according to

[19] , wherein the concentration of the coupler is 10 mmol / L or less.

[0035]

[26] The reagent composition for measurement according to any one of

[19] to

[25] , wherein the concentration of the coupler is the concentration of the coupler in the reagent composition for measurement.

[0036]

[27] The reagent composition for measurement according to any one of

[19] to

[25] , wherein the concentration of the coupler is the concentration of the coupler in a mixture of the reagent composition for measurement and a sample.

[0037]

[28] The reagent composition for measurement according to any one of [5] to

[27] , wherein the reagent composition for measurement comprises a first reagent and a second reagent, the first reagent comprises a coupler and a cyclodextrin or an analog thereof, and the second reagent comprises a Trinder reagent and a peroxidase.

[0038]

[29] The reagent composition for measurement according to any one of [5] to

[27] , wherein the reagent composition for measurement comprises a first reagent and a second reagent, the first reagent comprises a coupler, a peroxidase, and a cyclodextrin or an analog thereof, and the second reagent comprises Trinder's reagent.

[0039]

[30] The reagent composition for measurement according to any one of [5] to

[27] , wherein the reagent composition for measurement comprises a first reagent and a second reagent, the first reagent comprises a Trinder reagent and a cyclodextrin or an analogue thereof, and the second reagent comprises a coupler and a peroxidase.

[0040]

[31] The reagent composition for measurement according to any one of [5] to

[27] , wherein the reagent composition for measurement comprises a first reagent and a second reagent, the first reagent comprises Trinder's reagent, peroxidase, and cyclodextrin or an analogue thereof, and the second reagent comprises a coupler.

[0041]

[32] The reagent composition for measurement according to any one of

[27] to

[31] , wherein the first reagent is added to the sample, and then the second reagent is added to the sample.

[0042]

[33] The reagent composition for measurement according to any one of

[27] to

[32] , wherein the first reagent and the second reagent are packaged separately.

[0043]

[34] The reagent composition for measurement according to any one of [1] to [4], wherein the color-developing reagent is a leuco dye.

[0044]

[35] The reagent composition for measurement according to

[34] , wherein the leuco dye is one selected from the group consisting of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 10-(carboxymethylaminocarbonyl)-3,7-bis(dimethylamino)phenothiazine sodium (DA-67), N-(carboxymethylaminocarbonyl)-4,4-bis(dimethylamino)biphenylamine (DA-64), and N,N,N',N',N'',N''-hexa(3-sulfopropyl)-4,4',4''-triaminotriphenylmethane 6Na (TPM-PS).

[0045]

[36] The reagent composition for measurement according to

[35] , wherein the leuco dye is DA-67.

[0046]

[37] The reagent composition for measurement according to any one of

[34] to

[36] , wherein the concentration of the leuco dye is 0.005 mmol / L or more and 10 mmol / L or less.

[0047]

[38] The reagent composition for measurement according to

[37] , wherein the concentration of the leuco dye is 0.01 mmol / L or more.

[0048]

[39] The reagent composition for measurement according to

[37] , wherein the concentration of the leuco dye is 0.02 mmol / L or more.

[0049]

[40] The reagent composition for measurement according to

[37] , wherein the concentration of the leuco dye is 0.05 mmol / L or more.

[0050]

[41] The reagent composition for measurement according to

[37] , wherein the concentration of the leuco dye is 5 mmol / L or less.

[0051]

[42] The reagent composition for measurement according to

[37] , wherein the concentration of the leuco dye is 3 mmol / L or less.

[0052]

[43] The reagent composition for measurement according to

[37] , wherein the concentration of the leuco dye is 1 mmol / L or less.

[0053]

[44] The reagent composition for measurement according to any one of

[37] to

[43] , wherein the concentration of the leuco dye is the concentration of the leuco dye in the reagent composition for measurement.

[0054]

[45] The reagent composition for measurement according to any one of

[37] to

[43] , wherein the concentration of the leuco dye is the concentration of the leuco dye in a mixture of the reagent composition for measurement and a sample.

[0055]

[46] The reagent composition for measurement according to any one of [1] to

[45] , wherein the cyclodextrin is at least one selected from the group consisting of α-cyclodextrin, β-cyclodextrin, and / or γ-cyclodextrin.

[0056]

[47] The reagent composition for measurement according to

[46] , wherein the cyclodextrin or an analog thereof is α-cyclodextrin or an analog thereof.

[0057]

[48] ​​The reagent composition for measurement according to

[46] , wherein the cyclodextrin or an analog thereof is β-cyclodextrin or an analog thereof.

[0058]

[49] The reagent composition for measurement according to

[46] , wherein the cyclodextrin or an analogue thereof is γ-cyclodextrin or an analogue thereof.

[0059]

[50] The reagent composition for measurement according to any one of [1] to

[45] , wherein the cyclodextrin analogue is at least one selected from the group consisting of methyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, and hydroxyethyl-β-cyclodextrin.

[0060]

[51] The reagent composition for measurement according to

[50] , wherein the cyclodextrin analogue is methyl-β-cyclodextrin.

[0061]

[52] The reagent composition for measurement according to

[50] , wherein the cyclodextrin analogue is hydroxypropyl-β-cyclodextrin.

[0062]

[53] The reagent composition for measurement according to

[50] , wherein the cyclodextrin analogue is hydroxyethyl-β-cyclodextrin.

[0063]

[54] The reagent composition for measurement according to any one of [1] to

[53] , wherein the concentration of cyclodextrin or an analogue thereof is 0.1 weight / volume % or more and 35 weight / volume % or less.

[0064]

[55] The reagent composition for measurement according to

[54] , wherein the concentration of cyclodextrin or an analogue thereof is 0.5 weight / volume % or more.

[0065]

[56] The reagent composition for measurement according to

[54] , wherein the concentration of cyclodextrin or an analogue thereof is 1.0 weight / volume % or more.

[0066]

[57] The reagent composition for measurement according to

[54] , wherein the concentration of cyclodextrin or an analogue thereof is 5.0 weight / volume % or more.

[0067]

[58] The reagent composition for measurement according to

[54] , wherein the concentration of cyclodextrin or an analogue thereof is 30.0 weight / volume % or less.

[0068]

[59] The reagent composition for measurement according to

[54] , wherein the concentration of cyclodextrin or an analogue thereof is 25.0 weight / volume % or less.

[0069]

[60] The reagent composition for measurement according to any one of

[54] to

[59] , wherein the concentration of the cyclodextrin or its analogue is the concentration of the cyclodextrin or its analogue in the reagent composition for measurement.

[0070]

[61] The reagent composition for measurement according to any one of

[54] to

[59] , wherein the concentration of the cyclodextrin or its analogue is the concentration of the cyclodextrin or its analogue in a mixed solution of the reagent composition for measurement and a sample.

[0071]

[62] The reagent composition for measurement according to any one of [1] to

[61] , wherein the substance in the sample is one selected from the group consisting of uric acid (UA), creatinine (CRE), triglyceride (TG), cholesterol (CHO), and glycated proteins.

[0072]

[63] The reagent composition for measurement according to

[62] , wherein the substance in the sample is a glycated protein.

[0073]

[64] The reagent composition for measurement according to

[62] , wherein the glycated protein is glycated albumin (GA).

[0074]

[65] The reagent composition for measurement according to any one of [1] to

[64] , further comprising ferrocyanide.

[0075]

[66] The reagent composition for measurement according to

[65] , wherein the ferrocyanide is at least one selected from the group consisting of potassium ferrocyanide and sodium ferrocyanide.

[0076]

[67] The reagent composition for measurement according to

[65] or

[66] , wherein the concentration of ferrocyanide is 0.001 mmol / L or more and 10 mmol / L or less.

[0077]

[68] The reagent composition for measurement according to

[67] , wherein the concentration of ferrocyanide is 0.005 mmol / L or more.

[0078]

[69] The reagent composition for measurement according to

[67] , wherein the concentration of ferrocyanide is 0.01 mmol / L or more.

[0079]

[70] The reagent composition for measurement according to

[67] , wherein the concentration of ferrocyanide is 0.5 mmol / L or less.

[0080]

[71] The reagent composition for measurement according to

[67] , wherein the concentration of ferrocyanide is 0.1 mmol / L or less.

[0081]

[72] The concentration of the ferrocyanide is the concentration of the ferrocyanide in the reagent composition for measurement according to any one of

[67] to

[71] .

[0082]

[73] The reagent composition for measurement according to any one of

[67] to

[71] , wherein the concentration of the ferrocyanide is the concentration of the ferrocyanide in a mixture of the reagent composition for measurement and a sample.

[0083]

[74] The reagent composition for measurement according to any one of [1] to

[73] , further comprising a saccharide other than cyclodextrin or its analogue.

[0084]

[75] The reagent composition for measurement according to

[74] , wherein the saccharide other than cyclodextrin or its analogue is at least one selected from the group consisting of sugar alcohols, aldoses, ketoses, non-reducing disaccharides, and reducing disaccharides.

[0085]

[76] The reagent composition for measurement according to

[75] , wherein the sugar alcohol is at least one selected from the group consisting of erythritol, xylitol, sorbitol, and mannitol.

[0086]

[77] The reagent composition for measurement according to

[75] , wherein the aldose is at least one selected from the group consisting of erythrose, xylose, glucose, and mannose.

[0087]

[78] The reagent composition for measurement according to

[75] , wherein the ketose is at least one selected from the group consisting of erythrulose, xylulose, and fructose.

[0088]

[79] The reagent composition for measurement according to

[75] , wherein the non-reducing disaccharide is at least one selected from the group consisting of trehalose and sucrose.

[0089]

[80] The reagent composition for measurement according to

[75] , wherein the reducing disaccharide is maltose.

[0090]

[81] The reagent composition for measurement according to

[76] , wherein the sugar alcohol is at least one selected from the group consisting of sorbitol and mannitol.

[0091]

[82] The reagent composition for measurement according to

[81] , wherein the sugar alcohol is sorbitol.

[0092]

[83] The reagent composition for measurement according to

[81] , wherein the sugar alcohol is mannitol.

[0093]

[84] The reagent composition for measurement according to any one of

[74] to

[83] , wherein the concentration of the sugar is 0.1 weight / volume % or more and 50.0 weight / volume % or less.

[0094]

[85] The reagent composition for measurement according to

[84] , wherein the concentration of the sugar is 0.5 weight / volume % or more.

[0095]

[86] The reagent composition for measurement according to

[84] , wherein the concentration of the sugar is 1.0 weight / volume % or more.

[0096]

[87] The reagent composition for measurement according to

[84] , wherein the concentration of the sugar is 40.0 weight / volume % or less.

[0097]

[88] The reagent composition for measurement according to

[84] , wherein the concentration of the sugar is 30.0 weight / volume % or less.

[0098]

[89] The reagent composition for measurement described in any one of

[84] to

[88] , wherein the concentration of the sugar is the concentration of the sugar in the reagent composition for measurement.

[0099]

[90] The reagent composition for measurement according to any one of

[84] to

[88] , wherein the concentration of the sugar is the concentration of the sugar in a mixed solution of the reagent composition for measurement and a sample.

[0100]

[91] The reagent composition for measurement according to any one of [1] to

[90] , wherein cyclodextrin or an analog thereof suppresses color development inhibition caused by 2,5-dihydroxybenzenesulfonic acid or a salt thereof that may be contained in a sample.

[0101]

[92] The reagent composition for the measurement according to

[91] , wherein the 2,5-dihydroxybenzenesulfonic acid or a salt thereof is at least one selected from the group consisting of calcium dobesilate and ethanesilate.

[0102]

[93] A method for measuring a substance in a sample, comprising: preparing a measurement reagent composition comprising peroxidase, a color-developing reagent configured to react with hydrogen peroxide derived from a substance in the sample in the presence of peroxidase to produce a color, and cyclodextrin or an analog thereof; mixing the measurement reagent composition with the sample; and measuring the color development.

[0103]

[94] The method for measuring a substance in a sample according to

[93] , wherein the measurement reagent composition contains an effective amount of peroxidase.

[0104]

[95] The method for measuring a substance in a sample according to

[93] or

[94] , wherein the measurement reagent composition contains an effective amount of a color-developing reagent.

[0105]

[96] A method for measuring a substance in a sample according to any one of

[93] to

[95] , wherein the measurement reagent composition contains an effective amount of cyclodextrin or an analogue thereof.

[0106]

[97] A method for measuring a substance in a sample according to any one of

[93] to

[96] , wherein the color-developing reagent comprises a Trinder reagent and a coupler.

[0107]

[98] Trinder's reagent is N-(3-sulfopropyl)aniline sodium monohydrate (HALPS), N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium monohydrate (TOPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (MAOS), N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (HDAPS), N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (HDAOS), N-ethyl-N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (DAPS), N-ethyl-N-(2-hydroxy- ...

[97] The method for measuring a substance in a sample according to the above

[97] , wherein the substance is one selected from the group consisting of N-ethyl-N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium (DAOS), N-ethyl-N-(3-sulfopropyl)-3-methoxyaniline sodium monohydrate (ALPS), N-ethyl-N-(3-sulfopropyl)-3-methoxyaniline sodium dihydrate (ADPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methoxyaniline sodium dihydrate (ADOS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine (TOOS), N,N-bis(4-sulfobutyl)-3-methylaniline disodium (TODB), and 3-hydroxy-2,4,6-triiodobenzoic acid (HTIB).

[0108]

[99] The method for measuring a substance in a sample according to

[98] , wherein the Trinder reagent is TODB.

[0109]

[0100] A method for measuring a substance in a sample described in any of

[97] to

[99] , wherein the concentration of the Trinder reagent is 0.1 mmol / L or more and 50 mmol / L or less.

[0110]

[0101] A method for measuring a substance in a sample described in

[0100] , wherein the concentration of the Trinder reagent is 0.2 mmol / L or more.

[0111]

[0102] A method for measuring a substance in a sample described in

[0100] , wherein the concentration of the Trinder reagent is 0.5 mmol / L or more.

[0112]

[0103] A method for measuring a substance in a sample described in

[0100] , wherein the concentration of the Trinder reagent is 1.0 mmol / L or more.

[0113]

[0104] A method for measuring a substance in a sample described in

[0100] , wherein the concentration of the Trinder reagent is 30 mmol / L or less.

[0114]

[0105] A method for measuring a substance in a sample described in

[0100] , wherein the concentration of the Trinder reagent is 20 mmol / L or less.

[0115]

[0106] A method for measuring a substance in a sample described in

[0100] , wherein the concentration of the Trinder reagent is 10 mmol / L or less.

[0116]

[0107] A method for measuring a substance in a sample described in any one of

[0100] to

[0106] , wherein the concentration of the Trinder reagent is the concentration of the Trinder reagent in a mixed solution of the measurement reagent composition and the sample.

[0117]

[0108] A method for measuring a substance in a sample described in any of

[97] to

[0107] , wherein the coupler is one selected from the group consisting of 4-aminoantipyrine (4-AA), aminoantipyrine derivatives, vanillindiamine sulfonic acid, methylbenzthiazolinone hydrazone (MBTH), and sulfonated methylbenzthiazolinone hydrazone (SMBTH).

[0118]

[0109] A method for measuring a substance in a sample described in

[0108] , wherein the coupler is 4-AA.

[0119]

[0110] A method for measuring a substance in a sample described in any one of

[97] to

[0109] , wherein the concentration of the coupler is 0.1 mmol / L or more and 50 mmol / L or less.

[0120]

[0111] A method for measuring a substance in a sample described in

[0110] , wherein the concentration of the coupler is 0.2 mmol / L or more.

[0121]

[0112] A method for measuring a substance in a sample described in

[0110] , wherein the concentration of the coupler is 0.5 mmol / L or more.

[0122]

[0113] A method for measuring a substance in a sample described in

[0110] , wherein the concentration of the coupler is 1.0 mmol / L or more.

[0123]

[0114] A method for measuring a substance in a sample described in

[0110] , wherein the concentration of the coupler is 30 mmol / L or less.

[0124]

[0115] A method for measuring a substance in a sample described in

[0110] , wherein the concentration of the coupler is 20 mmol / L or less.

[0125]

[0116] A method for measuring a substance in a sample described in

[0110] , wherein the concentration of the coupler is 10 mmol / L or less.

[0126]

[0117] A method for measuring a substance in a sample described in any one of

[0110] to

[0116] , wherein the concentration of the coupler is the concentration of the coupler in a mixed solution of the measurement reagent composition and the sample.

[0127]

[0118] A method for measuring a substance in a sample described in any of

[97] to

[0117] , wherein the measurement reagent composition comprises a first reagent and a second reagent, the first reagent comprises a coupler and a cyclodextrin or an analogue thereof, and the second reagent comprises Trinder's reagent and peroxidase.

[0128]

[0119] A method for measuring a substance in a sample described in any of

[97] to

[0117] , wherein the measurement reagent composition comprises a first reagent and a second reagent, the first reagent comprises a coupler, a peroxidase, and a cyclodextrin or an analogue thereof, and the second reagent comprises Trinder's reagent.

[0129]

[0120] A method for measuring a substance in a sample described in any of

[97] to

[0117] , wherein the measurement reagent composition comprises a first reagent and a second reagent, the first reagent comprises Trinder's reagent and cyclodextrin or an analogue thereof, and the second reagent comprises a coupler and peroxidase.

[0130]

[0121] A method for measuring a substance in a sample described in any of

[97] to

[0117] , wherein the measurement reagent composition comprises a first reagent and a second reagent, the first reagent comprises Trinder's reagent, peroxidase, and cyclodextrin or an analogue thereof, and the second reagent comprises a coupler.

[0131]

[0122] A method for measuring a substance in a sample described in any one of

[0118] to

[0121] , wherein a first reagent and the sample are mixed, and then a second reagent and the sample are mixed.

[0132]

[0123] A method for measuring a substance in a sample described in any one of

[0118] to

[0122] , wherein the first reagent and the second reagent are packaged separately in the measurement reagent composition.

[0133]

[0124] A method for measuring a substance in a sample described in any one of

[93] to

[96] , wherein the color-developing reagent is a leuco dye.

[0134]

[0125] A method for measuring a substance in a sample described in

[0124] , wherein the leuco dye is one selected from the group consisting of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 10-(carboxymethylaminocarbonyl)-3,7-bis(dimethylamino)phenothiazine sodium (DA-67), N-(carboxymethylaminocarbonyl)-4,4-bis(dimethylamino)biphenylamine (DA-64), and N,N,N',N',N'',N''-hexa(3-sulfopropyl)-4,4',4''-triaminotriphenylmethane 6Na (TPM-PS).

[0135]

[0126] A method for measuring a substance in a sample described in

[0125] , wherein the leuco dye is DA-67.

[0136]

[0127] A method for measuring a substance in a sample described in any one of

[0124] to

[0126] , wherein the concentration of the leuco dye is 0.005 mmol / L or more and 10 mmol / L or less.

[0137]

[0128] A method for measuring a substance in a sample described in

[0127] , wherein the concentration of the leuco dye is 0.01 mmol / L or more.

[0138]

[0129] A method for measuring a substance in a sample described in

[0127] , wherein the concentration of the leuco dye is 0.02 mmol / L or more.

[0139]

[0130] A method for measuring a substance in a sample according to

[0127] , wherein the concentration of the leuco dye is 0.05 mmol / L or more.

[0140]

[0131] A method for measuring a substance in a sample described in

[0127] , wherein the concentration of the leuco dye is 5 mmol / L or less.

[0141]

[0132] A method for measuring a substance in a sample described in

[0127] , wherein the concentration of the leuco dye is 3 mmol / L or less.

[0142]

[0133] A method for measuring a substance in a sample described in

[0127] , wherein the concentration of the leuco dye is 1 mmol / L or less.

[0143]

[0134] A method for measuring a substance in a sample described in any one of

[0127] to

[0133] , wherein the concentration of the leuco dye is the concentration of the leuco dye in a mixed solution of the measurement reagent composition and the sample.

[0144]

[0135] A method for measuring a substance in a sample described in any one of

[93] to

[0134] , wherein the cyclodextrin is at least one selected from the group consisting of α-cyclodextrin, β-cyclodextrin, and / or γ-cyclodextrin.

[0145]

[0136] A method for measuring a substance in a sample described in

[0135] , wherein the cyclodextrin or an analog thereof is α-cyclodextrin or an analog thereof.

[0146]

[0137] A method for measuring a substance in a sample described in

[0135] , wherein the cyclodextrin or an analogue thereof is β-cyclodextrin or an analogue thereof.

[0147]

[0138] A method for measuring a substance in a sample described in

[0135] , wherein the cyclodextrin or an analog thereof is γ-cyclodextrin or an analog thereof.

[0148]

[0139] A method for measuring a substance in a sample described in any of

[93] to

[0134] , wherein the cyclodextrin analogue is at least one selected from the group consisting of methyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, and hydroxyethyl-β-cyclodextrin.

[0149]

[0140] A method for measuring a substance in a sample described in

[0139] , wherein the cyclodextrin analogue is methyl-β-cyclodextrin.

[0150]

[0141] A method for measuring a substance in a sample described in

[0139] , wherein the cyclodextrin analogue is hydroxypropyl-β-cyclodextrin.

[0151]

[0142] A method for measuring a substance in a sample described in

[0139] , wherein the cyclodextrin analogue is hydroxyethyl-β-cyclodextrin.

[0152]

[0143] A method for measuring a substance in a sample described in any one of

[93] to

[0142] , wherein the concentration of cyclodextrin or its analogue is 0.1 weight / volume % or more and 35 weight / volume % or less.

[0153]

[0144] A method for measuring a substance in a sample described in

[0143] , wherein the concentration of cyclodextrin or its analogue is 0.5 weight / volume % or more.

[0154]

[0145] A method for measuring a substance in a sample described in

[0143] , wherein the concentration of cyclodextrin or its analogue is 1.0 weight / volume % or more.

[0155]

[0146] A method for measuring a substance in a sample described in

[0143] , wherein the concentration of cyclodextrin or its analogue is 5.0 weight / volume % or more.

[0156]

[0147] A method for measuring a substance in a sample described in

[0143] , wherein the concentration of cyclodextrin or its analogue is 30.0 weight / volume % or less.

[0157]

[0148] A method for measuring a substance in a sample described in

[0143] , wherein the concentration of cyclodextrin or its analogue is 25.0 weight / volume % or less.

[0158]

[0149] A method for measuring a substance in a sample described in any one of

[0143] to

[0148] , wherein the concentration of the cyclodextrin or its analogue is the concentration of the cyclodextrin or its analogue in a mixed solution of the measurement reagent composition and the sample.

[0159]

[0150] A method for measuring a substance in a sample described in any of

[93] to

[0149] , wherein the substance in the sample is one selected from the group consisting of uric acid (UA), creatinine (CRE), triglyceride (TG), cholesterol (CHO), and glycated proteins.

[0160]

[0151] A method for measuring a substance in a sample described in

[0150] , wherein the substance in the sample is a glycated protein.

[0161]

[0152] A method for measuring a substance in a sample described in

[0150] , wherein the glycated protein is glycated albumin (GA).

[0162]

[0153] A method for measuring a substance in a sample described in any one of

[93] to

[0152] , wherein the measurement reagent composition further comprises ferrocyanide.

[0163]

[0154] A method for measuring a substance in a sample described in

[0153] , wherein the ferrocyanide is at least one selected from the group consisting of potassium ferrocyanide and sodium ferrocyanide.

[0164]

[0155] A method for measuring a substance in a sample described in

[0153] or

[0154] , wherein the concentration of ferrocyanide is 0.001 mmol / L or more and 10 mmol / L or less.

[0165]

[0156] A method for measuring a substance in a sample described in

[0155] , wherein the concentration of ferrocyanide is 0.005 mmol / L or more.

[0166]

[0157] A method for measuring a substance in a sample described in

[0155] , wherein the concentration of ferrocyanide is 0.01 mmol / L or more.

[0167]

[0158] A method for measuring a substance in a sample described in

[0155] , wherein the concentration of ferrocyanide is 0.5 mmol / L or less.

[0168]

[0159] A method for measuring a substance in a sample described in

[0155] , wherein the concentration of ferrocyanide is 0.1 mmol / L or less.

[0169]

[0160] A method for measuring a substance in a sample described in any one of

[0155] to

[0159] , wherein the concentration of the ferrocyanide is the concentration of the ferrocyanide in a mixed solution of the measurement reagent composition and the sample.

[0170]

[0161] A method for measuring a substance in a sample described in any one of

[93] to

[0160] , wherein the measurement reagent composition further comprises a saccharide other than cyclodextrin or its analogue.

[0171]

[0162] A method for measuring a substance in a sample described in

[0161] , wherein the sugar other than cyclodextrin or its analogue is at least one selected from the group consisting of sugar alcohols, aldoses, ketoses, non-reducing disaccharides, and reducing disaccharides.

[0172]

[0163] A method for measuring a substance in a sample described in

[0162] , wherein the sugar alcohol is at least one selected from the group consisting of erythritol, xylitol, sorbitol, and mannitol.

[0173]

[0164] A method for measuring a substance in a sample described in

[0162] , wherein the aldose is at least one selected from the group consisting of erythrose, xylose, glucose, and mannose.

[0174]

[0165] A method for measuring a substance in a sample described in

[0162] , wherein the ketose is at least one selected from the group consisting of erythrulose, xylulose, and fructose.

[0175]

[0166] A method for measuring a substance in a sample described in

[0162] , wherein the non-reducing disaccharide is at least one selected from the group consisting of trehalose and sucrose.

[0176]

[0167] A method for measuring a substance in a sample described in

[0162] , wherein the reducing disaccharide is maltose.

[0177]

[0168] A method for measuring a substance in a sample described in

[0163] , wherein the sugar alcohol is at least one selected from the group consisting of sorbitol and mannitol.

[0178]

[0169] A method for measuring a substance in a sample described in

[0168] , wherein the sugar alcohol is sorbitol.

[0179]

[0170] A method for measuring a substance in a sample described in

[0168] , wherein the sugar alcohol is mannitol.

[0180]

[0171] A method for measuring a substance in a sample described in

[0161] to

[0170] , wherein the sugar concentration is 0.1 weight / volume% or more and 50.0 weight / volume% or less.

[0181]

[0172] A method for measuring a substance in a sample described in

[0171] , wherein the sugar concentration is 0.5 weight / volume % or more.

[0182]

[0173] A method for measuring a substance in a sample described in

[0171] , wherein the sugar concentration is 1.0 weight / volume % or more.

[0183]

[0174] A method for measuring a substance in a sample described in

[0171] , wherein the sugar concentration is 40.0 weight / volume % or less.

[0184]

[0175] A method for measuring a substance in a sample described in

[0171] , wherein the sugar concentration is 30.0 weight / volume % or less.

[0185]

[0176] A method for measuring a substance in a sample described in any one of

[0171] to

[0175] , wherein the concentration of the sugar is the concentration of the sugar in a mixed solution of the measurement reagent composition and the sample.

[0186]

[0177] A method for measuring a substance in a sample described in any one of

[93] to

[0176] , in which cyclodextrin or an analogue thereof suppresses color inhibition caused by 2,5-dihydroxybenzenesulfonic acid or a salt thereof that may be contained in the sample.

[0187]

[0178] A method for measuring a substance in a sample described in

[0177] , wherein 2,5-dihydroxybenzenesulfonic acid or a salt thereof is at least one selected from the group consisting of calcium dobesilate and ethanesilate.

[0188]

[0179] Use of cyclodextrin or an analogue thereof as an inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof.

[0189]

[0180] Use of cyclodextrin or an analog thereof in the production of an inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof.

[0190]

[0181] Use of cyclodextrin or an analogue thereof as an inhibitor of color development inhibition by 2,5-dihydroxybenzenesulfonic acid or a salt thereof in a measurement reagent composition comprising peroxidase and a color-developing reagent configured to react with hydrogen peroxide derived from a substance in a sample in the presence of peroxidase to develop color.

[0191]

[0182] Use of cyclodextrin or an analogue thereof in the manufacture of an inhibitor of color development inhibition by 2,5-dihydroxybenzenesulfonic acid or a salt thereof in a measurement reagent composition comprising peroxidase and a color-developing reagent configured to react with hydrogen peroxide derived from a substance in a sample in the presence of peroxidase to develop color.

[0192]

[0183] The use described in

[0181] or

[0182] , wherein the measurement reagent composition contains an effective amount of peroxidase.

[0193]

[0184] A use described in any of

[0181] to

[0183] , wherein the measurement reagent composition contains an effective amount of a color-developing reagent.

[0194]

[0185] The use described in any one of

[0181] to

[0184] , wherein the measurement reagent composition contains an effective amount of cyclodextrin or an analogue thereof.

[0195]

[0186] The use described in any one of

[0181] to

[0185] , wherein the color-developing reagent comprises a Trinder reagent and a coupler.

[0196]

[0187] Trinder's reagent is N-(3-sulfopropyl)aniline sodium monohydrate (HALPS), N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium monohydrate (TOPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (MAOS), N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (HDAPS), N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (HDAOS), N-ethyl-N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (DAPS), N-ethyl-N-(2-hydroxy-3- The use according to

[0186] , wherein the is one selected from the group consisting of N-ethyl-N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium (DAOS), N-ethyl-N-(3-sulfopropyl)-aniline sodium (ALPS), N-ethyl-N-(3-sulfopropyl)-3-methoxyaniline sodium monohydrate (ADPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methoxyaniline sodium dihydrate (ADOS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine (TOOS), N,N-bis(4-sulfobutyl)-3-methylaniline disodium (TODB), and 3-hydroxy-2,4,6-triiodobenzoic acid (HTIB).

[0197]

[0188] The use described in

[0187] , wherein the Trinder reagent is TODB.

[0198]

[0189] The use described in any of

[0186] to

[0188] , wherein the concentration of the Trinder reagent is 0.1 mmol / L or more and 50 mmol / L or less.

[0199]

[0190] The use described in

[0189] , wherein the concentration of the Trinder reagent is 0.2 mmol / L or more.

[0200]

[0191] The use described in

[0189] , wherein the concentration of the Trinder reagent is 0.5 mmol / L or more.

[0201]

[0192] The use described in

[0189] , wherein the concentration of the Trinder reagent is 1.0 mmol / L or more.

[0202]

[0193] The use described in

[0189] , wherein the concentration of the Trinder reagent is 30 mmol / L or less.

[0203]

[0194] The use described in

[0189] , wherein the concentration of the Trinder reagent is 20 mmol / L or less.

[0204]

[0195] The use described in

[0189] , wherein the concentration of the Trinder reagent is 10 mmol / L or less.

[0205]

[0196] The use described in any one of

[0189] to

[0195] , wherein the concentration of the Trinder reagent is the concentration of the Trinder reagent in a mixed solution of the measurement reagent composition and the sample.

[0206]

[0197] The use described in any one of

[0186] to

[0196] , wherein the coupler is one selected from the group consisting of 4-aminoantipyrine (4-AA), aminoantipyrine derivatives, vanillin diamine sulfonic acid, methyl benzthiazolinone hydrazone (MBTH), and sulfonated methyl benzthiazolinone hydrazone (SMBTH).

[0207]

[0198] The use described in

[0197] , wherein the coupler is 4-AA.

[0208]

[0199] The use described in any one of

[0186] to

[0198] , wherein the concentration of the coupler is 0.1 mmol / L or more and 50 mmol / L or less.

[0209]

[0200] The use described in

[0199] , wherein the concentration of the coupler is 0.2 mmol / L or more.

[0210]

[0201] The use described in

[0199] , wherein the concentration of the coupler is 0.5 mmol / L or more.

[0211]

[0202] The use described in

[0199] , wherein the concentration of the coupler is 1.0 mmol / L or more.

[0212]

[0203] The use described in

[0199] , wherein the concentration of the coupler is 30 mmol / L or less.

[0213]

[0204] The use described in

[0199] , wherein the concentration of the coupler is 20 mmol / L or less.

[0214]

[0205] The use described in

[0199] , wherein the concentration of the coupler is 10 mmol / L or less.

[0215]

[0206] The use described in any one of

[0199] to

[0205] , wherein the concentration of the coupler is the concentration of the coupler in a mixed solution of the measurement reagent composition and the sample.

[0216]

[0207] The use described in any of

[0186] to

[0206] , wherein the measurement reagent composition comprises a first reagent and a second reagent, the first reagent comprises a coupler and a cyclodextrin or an analogue thereof, and the second reagent comprises a Trinder reagent and peroxidase.

[0217]

[0208] The use described in any of

[0186] to

[0206] , wherein the measurement reagent composition comprises a first reagent and a second reagent, the first reagent comprises a coupler, a peroxidase, and a cyclodextrin or an analogue thereof, and the second reagent comprises Trinder's reagent.

[0218]

[0209] The use described in any of

[0186] to

[0206] , wherein the measurement reagent composition comprises a first reagent and a second reagent, the first reagent comprises Trinder's reagent and cyclodextrin or an analogue thereof, and the second reagent comprises a coupler and peroxidase.

[0219]

[0210] The use described in any of

[0186] to

[0206] , wherein the measurement reagent composition comprises a first reagent and a second reagent, the first reagent comprises Trinder's reagent, peroxidase, and cyclodextrin or an analogue thereof, and the second reagent comprises a coupler.

[0220]

[0211] A use described in any of

[0207] to

[0210] , wherein the first reagent and the sample are mixed and then the second reagent and the sample are mixed.

[0221]

[0212] A use described in any of

[0207] to

[0211] , wherein the first reagent and the second reagent are packaged separately in the measurement reagent composition.

[0222]

[0213] The use described in any one of

[0181] to

[0185] , wherein the color-developing reagent is a leuco dye.

[0223]

[0214] The use described in

[0213] , wherein the leuco dye is one selected from the group consisting of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 10-(carboxymethylaminocarbonyl)-3,7-bis(dimethylamino)phenothiazine sodium (DA-67), N-(carboxymethylaminocarbonyl)-4,4-bis(dimethylamino)biphenylamine (DA-64), and N,N,N',N',N'',N''-hexa(3-sulfopropyl)-4,4',4''-triaminotriphenylmethane 6Na (TPM-PS).

[0224]

[0215] The use according to

[0214] , wherein the leuco dye is DA-67.

[0225]

[0216] The use described in any one of

[0213] to

[0215] , wherein the concentration of the leuco dye is 0.005 mmol / L or more and 10 mmol / L or less.

[0226]

[0217] The use described in

[0216] , wherein the concentration of the leuco dye is 0.01 mmol / L or more.

[0227]

[0218] The use described in

[0216] , wherein the concentration of the leuco dye is 0.02 mmol / L or more.

[0228]

[0219] The use described in

[0216] , wherein the concentration of the leuco dye is 0.05 mmol / L or more.

[0229]

[0220] The use described in

[0216] , wherein the concentration of the leuco dye is 5 mmol / L or less.

[0230]

[0221] The use described in

[0216] , wherein the concentration of the leuco dye is 3 mmol / L or less.

[0231]

[0222] The use described in

[0216] , wherein the concentration of the leuco dye is 1 mmol / L or less.

[0232]

[0223] The use described in any one of

[0216] to

[0222] , wherein the concentration of the leuco dye is the concentration of the leuco dye in a mixed solution of the measurement reagent composition and a sample.

[0233]

[0224] The use described in any one of

[0179] to

[0223] , wherein the cyclodextrin is at least one selected from the group consisting of α-cyclodextrin, β-cyclodextrin, and / or γ-cyclodextrin.

[0234]

[0225] The use described in

[0224] , wherein the cyclodextrin or an analog thereof is α-cyclodextrin or an analog thereof.

[0235]

[0226] The use described in

[0224] , wherein the cyclodextrin or an analog thereof is β-cyclodextrin or an analog thereof.

[0236]

[0227] The use described in

[0224] , wherein the cyclodextrin or an analog thereof is γ-cyclodextrin or an analog thereof.

[0237]

[0228] The use described in any one of

[0179] to

[0223] , wherein the cyclodextrin analog is at least one selected from the group consisting of methyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, and hydroxyethyl-β-cyclodextrin.

[0238]

[0229] The use described in

[0228] , wherein the cyclodextrin analog is methyl-β-cyclodextrin.

[0239]

[0230] The use described in

[0228] , wherein the cyclodextrin analog is hydroxypropyl-β-cyclodextrin.

[0240]

[0231] The use described in

[0228] , wherein the cyclodextrin analog is hydroxyethyl-β-cyclodextrin.

[0241]

[0232] The use described in any one of

[0179] to

[0231] , wherein the concentration of cyclodextrin or an analogue thereof is 0.1 weight / volume % or more and 35 weight / volume % or less.

[0242]

[0233] The use described in

[0232] , wherein the concentration of cyclodextrin or its analogue is 0.5 weight / volume % or more.

[0243]

[0234] The use described in

[0232] , wherein the concentration of cyclodextrin or an analogue thereof is 1.0 weight / volume % or more.

[0244]

[0235] The use described in

[0232] , wherein the concentration of cyclodextrin or its analogue is 5.0 weight / volume % or more.

[0245]

[0236] The use described in

[0232] , wherein the concentration of cyclodextrin or its analogue is 30.0 weight / volume % or less.

[0246]

[0237] The use described in

[0232] , wherein the concentration of cyclodextrin or its analogue is 25.0 weight / volume % or less.

[0247]

[0238] The use described in any one of

[0232] to

[0237] , wherein the concentration of the cyclodextrin or its analogue is the concentration of the cyclodextrin or its analogue in a mixed solution of the measurement reagent composition and a sample.

[0248]

[0239] The use described in any of

[0181] to

[0223] , wherein the substance in the sample is one selected from the group consisting of uric acid (UA), creatinine (CRE), triglyceride (TG), cholesterol (CHO), and glycated proteins.

[0249]

[0240] The use described in

[0239] , wherein the substance in the sample is a glycated protein.

[0250]

[0241] The use described in

[0239] , wherein the glycated protein is glycated albumin (GA).

[0251]

[0242] The use described in any one of

[0181] to

[0223] ,

[0240] and

[0241] , wherein the measurement reagent composition further comprises ferrocyanide.

[0252]

[0243] The use described in

[0242] , wherein the ferrocyanide is at least one selected from the group consisting of potassium ferrocyanide and sodium ferrocyanide.

[0253]

[0244] The use described in

[0242] or

[0243] , wherein the concentration of ferrocyanide is 0.001 mmol / L or more and 10 mmol / L or less.

[0254]

[0245] The use described in

[0244] , wherein the concentration of ferrocyanide is 0.005 mmol / L or more.

[0255]

[0246] The use described in

[0244] , wherein the concentration of ferrocyanide is 0.01 mmol / L or more.

[0256]

[0247] The use described in

[0244] , wherein the concentration of ferrocyanide is 0.5 mmol / L or less.

[0257]

[0248] The use described in

[0244] , wherein the concentration of ferrocyanide is 0.1 mmol / L or less.

[0258]

[0249] The use described in any one of

[0244] to

[0248] , wherein the concentration of the ferrocyanide is the concentration of the ferrocyanide in a mixture of the measurement reagent composition and the sample.

[0259]

[0250] A use described in any one of

[0181] to

[0223] and

[0240] to

[0249] , wherein the measurement reagent composition further comprises a sugar other than cyclodextrin or an analogue thereof.

[0260]

[0251] The use described in

[0250] , wherein the sugar other than cyclodextrin or an analog thereof is at least one selected from the group consisting of sugar alcohols, aldoses, ketoses, non-reducing disaccharides, and reducing disaccharides.

[0261]

[0252] The use described in

[0251] , wherein the sugar alcohol is at least one selected from the group consisting of erythritol, xylitol, sorbitol, and mannitol.

[0262]

[0253] The use described in

[0251] , wherein the aldose is at least one selected from the group consisting of erythrose, xylose, glucose, and mannose.

[0263]

[0254] The use described in

[0251] , wherein the ketose is at least one selected from the group consisting of erythrulose, xylulose, and fructose.

[0264]

[0255] The use described in

[0251] , wherein the non-reducing disaccharide is at least one selected from the group consisting of trehalose and sucrose.

[0265]

[0256] The use described in

[0251] , wherein the reducing disaccharide is maltose.

[0266]

[0257] The use described in

[0252] , wherein the sugar alcohol is at least one selected from the group consisting of sorbitol and mannitol.

[0267]

[0258] The use described in

[0257] , wherein the sugar alcohol is sorbitol.

[0268]

[0259] The use described in

[0257] , wherein the sugar alcohol is mannitol.

[0269]

[0260] The use described in

[0250] to

[0259] , wherein the concentration of the sugar is 0.1 weight / volume % or more and 50.0 weight / volume % or less.

[0270]

[0261] The use described in

[0260] , wherein the concentration of sugar is 0.5 weight / volume % or more.

[0271]

[0262] The use described in

[0260] , wherein the concentration of sugar is 1.0 weight / volume % or more.

[0272]

[0263] The use described in

[0260] , wherein the sugar concentration is 40.0 weight / volume % or less.

[0273]

[0264] The use described in

[0260] , wherein the sugar concentration is 30.0 weight / volume % or less.

[0274]

[0265] The use described in any one of

[0260] to

[0264] , wherein the concentration of the sugar is the concentration of the sugar in a mixed solution of the measurement reagent composition and the sample.

[0275]

[0266] The use described in any one of

[0181] to

[0223] and

[0240] to

[0265] , wherein cyclodextrin or an analog thereof suppresses color inhibition caused by 2,5-dihydroxybenzenesulfonic acid or a salt thereof that may be contained in a sample.

[0276]

[0267] The use described in any of

[0179] to

[0266] , wherein cyclodextrin or an analog thereof inhibits the action of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on a substance.

[0277]

[0268] The use described in any one of

[0179] to

[0267] , wherein cyclodextrin or an analogue thereof inhibits the action of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on a color-developing reagent.

[0278]

[0269] The use described in any one of

[0179] to

[0268] , wherein cyclodextrin or an analog thereof inhibits the inhibition of a color-developing reaction using a color-developing reagent by 2,5-dihydroxybenzenesulfonic acid or a salt thereof.

[0279]

[0270] The use described in any of

[0179] to

[0269] , wherein 2,5-dihydroxybenzenesulfonic acid or a salt thereof is at least one selected from the group consisting of calcium dobesilate and ethanesilate.

[0280]

[0271] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or its salt, including cyclodextrin or an analogue thereof.

[0281]

[0272] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in

[0271] , which contains an effective amount of cyclodextrin or an analogue thereof.

[0282]

[0273] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0271] or

[0272] , which inhibits the action of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on a substance.

[0283]

[0274] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0271] or

[0272] , which inhibits the action of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on a color-developing reagent.

[0284]

[0275] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0271] or

[0272] , which inhibits the inhibition of a color-developing reaction using a color-developing reagent by 2,5-dihydroxybenzenesulfonic acid or a salt thereof.

[0285]

[0276] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0274] or

[0275] , wherein the color-developing reagent reacts with hydrogen peroxide in the presence of peroxidase to develop color.

[0286]

[0277] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0274] to

[0276] , wherein the color-developing reagent is at least one selected from the group consisting of a Trinder reagent and a coupler.

[0287]

[0278] Trinder's reagent is N-(3-sulfopropyl)aniline sodium monohydrate (HALPS), N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium monohydrate (TOPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (MAOS), N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (HDAPS), N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (HDAOS), N-ethyl-N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (DAPS), N-ethyl-N-(2-hydroxy- ... The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0277] , which is one selected from the group consisting of dimethoxyaniline sodium (DAOS), N-ethyl-N-(3-sulfopropyl)aniline sodium (ALPS), N-ethyl-N-(3-sulfopropyl)-3-methoxyaniline sodium monohydrate (ADPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methoxyaniline sodium dihydrate (ADOS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine (TOOS), N,N-bis(4-sulfobutyl)-3-methylaniline disodium (TODB), and 3-hydroxy-2,4,6-triiodobenzoic acid (HTIB).

[0288]

[0279] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0278] , wherein the Trinder reagent is TODB.

[0289]

[0280] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0277] to

[0279] , wherein the coupler is one selected from the group consisting of 4-aminoantipyrine (4-AA), aminoantipyrine derivatives, vanillindiamine sulfonic acid, methylbenzthiazolinone hydrazone (MBTH), and sulfonated methylbenzthiazolinone hydrazone (SMBTH).

[0290]

[0281] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0280] , wherein the coupler is 4-AA.

[0291]

[0282] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0274] to

[0276] , wherein the color-developing reagent is a leuco dye.

[0292]

[0283] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0282] , wherein the leuco dye is one selected from the group consisting of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 10-(carboxymethylaminocarbonyl)-3,7-bis(dimethylamino)phenothiazine sodium (DA-67), N-(carboxymethylaminocarbonyl)-4,4-bis(dimethylamino)biphenylamine (DA-64), and N,N,N',N',N'',N''-hexa(3-sulfopropyl)-4,4',4''-triaminotriphenylmethane 6Na (TPM-PS).

[0293]

[0284] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0283] , wherein the leuco dye is DA-67.

[0294]

[0285] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0271] to

[0284] , which inhibits the action of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on peroxidase.

[0295]

[0286] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0271] to

[0284] , which inhibits the action of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on hydrogen peroxide.

[0296]

[0287] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any of

[0271] to

[0286] , wherein 2,5-dihydroxybenzenesulfonic acid or a salt thereof is at least one selected from the group consisting of calcium dobesilate and ethamsylate.

[0297]

[0288] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0271] to

[0287] , wherein the cyclodextrin is at least one selected from the group consisting of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin.

[0298]

[0289] The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0288] , wherein the cyclodextrin or an analogue thereof is α-cyclodextrin or an analogue thereof.

[0299]

[0290] The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0288] , wherein the cyclodextrin or an analogue thereof is β-cyclodextrin or an analogue thereof.

[0300]

[0291] The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0288] , wherein the cyclodextrin or an analogue thereof is γ-cyclodextrin or an analogue thereof.

[0301]

[0292] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0271] to

[0287] , wherein the cyclodextrin analogue is at least one selected from the group consisting of methyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, and hydroxyethyl-β-cyclodextrin.

[0302]

[0293] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0292] , wherein the cyclodextrin analogue is methyl-β-cyclodextrin.

[0303]

[0294] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0292] , wherein the cyclodextrin analogue is hydroxypropyl-β-cyclodextrin.

[0304]

[0295] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0292] , wherein the cyclodextrin analogue is hydroxyethyl-β-cyclodextrin.

[0305]

[0296] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0271] to

[0295] , further comprising ferrocyanide.

[0306]

[0297] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0296] , wherein the ferrocyanide is at least one selected from the group consisting of potassium ferrocyanide and sodium ferrocyanide.

[0307]

[0298] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0271] to

[0297] , further comprising a sugar other than cyclodextrin or an analogue thereof.

[0308]

[0299] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in

[0298] , wherein the sugar other than cyclodextrin or an analogue thereof is at least one selected from the group consisting of sugar alcohols, aldoses, ketoses, non-reducing disaccharides, and reducing disaccharides.

[0309]

[0300] The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in

[0299] , wherein the sugar alcohol is at least one selected from the group consisting of erythritol, xylitol, sorbitol, and mannitol.

[0310]

[0301] The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0299] , wherein the aldose is at least one selected from the group consisting of erythrose, xylose, glucose, and mannose.

[0311]

[0302] The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in

[0299] , wherein the ketose is at least one selected from the group consisting of erythrulose, xylulose, and fructose.

[0312]

[0303] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0299] , wherein the non-reducing disaccharide is at least one selected from the group consisting of trehalose and sucrose.

[0313]

[0304] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0299] , wherein the reducing disaccharide is maltose.

[0314]

[0305] The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0300] , wherein the sugar alcohol is at least one selected from the group consisting of sorbitol and mannitol.

[0315]

[0306] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0305] , wherein the sugar alcohol is sorbitol.

[0316]

[0307] An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0305] , wherein the sugar alcohol is mannitol.

[0317]

[0308] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof, comprising mixing cyclodextrin or an analogue thereof with 2,5-dihydroxybenzenesulfonic acid or a salt thereof.

[0318]

[0309] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in

[0308] , which comprises mixing an effective amount of cyclodextrin or an analogue thereof with 2,5-dihydroxybenzenesulfonic acid or a salt thereof.

[0319]

[0310] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0308] or

[0309] , which inhibits the action of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on a substance.

[0320]

[0311] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0308] or

[0309] , which inhibits the action of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on a color-developing reagent.

[0321]

[0312] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0308] or

[0309] , which inhibits 2,5-dihydroxybenzenesulfonic acid or a salt thereof from inhibiting a color-developing reaction using a color-developing reagent.

[0322]

[0313] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0311] or

[0312] , wherein the color-developing reagent is a color-developing reagent that reacts with hydrogen peroxide in the presence of peroxidase to develop color.

[0323]

[0314] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0311] to

[0313] , wherein the color-developing reagent is at least one selected from the group consisting of Trinder's reagent and a coupler.

[0324]

[0315] Trinder's reagent is N-(3-sulfopropyl)aniline sodium monohydrate (HALPS), N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium monohydrate (TOPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (MAOS), N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (HDAPS), N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (HDAOS), N-ethyl-N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (DAPS), N-ethyl-N-(2-hydroxy- ... The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0314] , wherein the compound is one selected from the group consisting of N-ethyl-N-(3-sulfopropyl)aniline sodium (DAOS), N-ethyl-N-(3-sulfopropyl)-3-methoxyaniline sodium monohydrate (ADPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methoxyaniline sodium dihydrate (ADOS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine (TOOS), N,N-bis(4-sulfobutyl)-3-methylaniline disodium (TODB), and 3-hydroxy-2,4,6-triiodobenzoic acid (HTIB).

[0325]

[0316] The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0315] , wherein the Trinder reagent is TODB.

[0326]

[0317] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0314] to

[0316] , wherein the coupler is one selected from the group consisting of 4-aminoantipyrine (4-AA), aminoantipyrine derivatives, vanillindiaminesulfonic acid, methylbenzthiazolinone hydrazone (MBTH), and sulfonated methylbenzthiazolinone hydrazone (SMBTH).

[0327]

[0318] The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0317] , wherein the coupler is 4-AA.

[0328]

[0319] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to any one of

[0311] to

[0313] , wherein the color-developing reagent is a leuco dye.

[0329]

[0320] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0319] , wherein the leuco dye is one selected from the group consisting of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 10-(carboxymethylaminocarbonyl)-3,7-bis(dimethylamino)phenothiazine sodium (DA-67), N-(carboxymethylaminocarbonyl)-4,4-bis(dimethylamino)biphenylamine (DA-64), and N,N,N',N',N'',N''-hexa(3-sulfopropyl)-4,4',4''-triaminotriphenylmethane 6Na (TPM-PS).

[0330]

[0321] The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0320] , wherein the leuco dye is DA-67.

[0331]

[0322] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0308] to

[0321] , wherein 2,5-dihydroxybenzenesulfonic acid or a salt thereof is at least one selected from the group consisting of calcium dobesilate and ethanesilate.

[0332]

[0323] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0308] to

[0322] , wherein the cyclodextrin is at least one selected from the group consisting of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin.

[0333]

[0324] The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0323] , wherein the cyclodextrin or an analogue thereof is α-cyclodextrin or an analogue thereof.

[0334]

[0325] The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0323] , wherein the cyclodextrin or an analogue thereof is β-cyclodextrin or an analogue thereof.

[0335]

[0326] The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0323] , wherein the cyclodextrin or an analogue thereof is γ-cyclodextrin or an analogue thereof.

[0336]

[0327] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0308] to

[0322] , wherein the cyclodextrin analogue is at least one selected from the group consisting of methyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, and hydroxyethyl-β-cyclodextrin.

[0337]

[0328] The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0327] , wherein the cyclodextrin analogue is methyl-β-cyclodextrin.

[0338]

[0329] The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0327] , wherein the cyclodextrin analogue is hydroxypropyl-β-cyclodextrin.

[0339]

[0330] The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0327] , wherein the cyclodextrin analogue is hydroxyethyl-β-cyclodextrin.

[0340]

[0331] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0308] to

[0330] , in which ferrocyanide is further mixed in when mixing cyclodextrin or an analogue thereof with 2,5-dihydroxybenzenesulfonic acid or a salt thereof.

[0341]

[0332] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0331] , wherein the ferrocyanide is at least one selected from the group consisting of potassium ferrocyanide and sodium ferrocyanide.

[0342]

[0333] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in any one of

[0308] to

[0332] , in which, in mixing cyclodextrin or an analogue thereof with 2,5-dihydroxybenzenesulfonic acid or a salt thereof, a sugar other than cyclodextrin or an analogue thereof is further mixed in.

[0343]

[0334] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof described in

[0333] , wherein the sugar other than cyclodextrin or an analogue thereof is at least one selected from the group consisting of sugar alcohols, aldoses, ketoses, non-reducing disaccharides, and reducing disaccharides.

[0344]

[0335] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0334] , wherein the sugar alcohol is at least one selected from the group consisting of erythritol, xylitol, sorbitol, and mannitol.

[0345]

[0336] The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0334] , wherein the aldose is at least one selected from the group consisting of erythrose, xylose, glucose, and mannose.

[0346]

[0337] The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0334] , wherein the ketose is at least one selected from the group consisting of erythrulose, xylulose, and fructose.

[0347]

[0338] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0334] , wherein the non-reducing disaccharide is at least one selected from the group consisting of trehalose and sucrose.

[0348]

[0339] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0334] , wherein the reducing disaccharide is maltose.

[0349]

[0340] The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0335] , wherein the sugar alcohol is at least one selected from the group consisting of sorbitol and mannitol.

[0350]

[0341] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0340] , wherein the sugar alcohol is sorbitol.

[0351]

[0342] A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to

[0340] , wherein the sugar alcohol is mannitol. Effect of the Invention

[0352] According to the present invention, it is possible to provide an inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof, and a reagent composition for sample measurement that suppresses the effect of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on measurement when measuring a substance in a sample. [Brief description of the drawings]

[0353] [Figure 1] 2 is a graph showing the results of one-dimensional NMR measurement in Example 1. [Diagram 2]1 is a graph showing the results of two-dimensional NMR measurement in Example 2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0354] A preferred embodiment of the present invention (hereinafter referred to as "embodiment") will be described in detail below. Note that the embodiment described below is an example of an apparatus and method for embodying the technical idea of ​​the present invention, and the technical idea of ​​the present invention is not limited to the combination of components described below. The technical idea of ​​the present invention can be modified in various ways within the scope of the claims.

[0355] The chemical formula for 2,5-dihydroxybenzenesulphonic acid is C 6 H 6 O 5 S and has the following structure: sulfonic acid SO 3 H is mainly reacted with SO in solution. 3 - It exists as an ion. [ka]

[0356] 2,5-Dihydroxybenzenesulfonic acid H + The salts in which the ions are exchanged with other cations are not particularly limited. 2,5-dihydroxybenzenesulfonic acid or its salts may be in the form of a simple substance or a hydrate. The salts may be metal salts. The salts may be amine salts. Examples of salts of 2,5-dihydroxybenzenesulfonic acid include calcium dobesilate and ethanesilate.

[0357] The chemical formula of calcium dobesilate, also known as 2,5-dihydroxybenzenesulfonic acid calcium salt or calcium 2,5-dihydroxybenzenesulfonate, is (C 6 H 5 O 5 S) 2 .Ca and has the following structure: [ka]

[0358] Calcium dobesilate may be a hydrate. Calcium dobesilate hydrate (2,5-dihydroxybenzenesulfonic acid calcium salt hydrate, or calcium 2,5-dihydroxybenzenesulfonate hydrate) has the following structure: [ka]

[0359] Diethylamine 2,5-dihydroxybenzenesulfonate, also known as ethanesylate or ethamsylate, has the chemical formula C 10 H 17 NO 5 S and has the following structure: [ka]

[0360] The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to the embodiment includes cyclodextrin or an analog thereof. The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to the embodiment includes an effective amount of cyclodextrin or an analog thereof. Alternatively, the inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to the embodiment consists of cyclodextrin or an analog thereof.

[0361] The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to the embodiment can be used as an inhibitor that inhibits the action of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on other substances. The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to the embodiment can be used as an inhibitor that inhibits the action of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on a color-developing reagent. In the present disclosure, the color-developing reagent includes a reagent that functions as a color-developing reagent by itself, such as a leuco-type dye, and a reagent that develops color by oxidative condensation of dye precursors, such as a Trinder reagent and a coupler. The color-developing reagent also includes a dye precursor by itself and a compound in which dye precursors are condensed by each other.

[0362] The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to the embodiment can be used as an inhibitor that inhibits 2,5-dihydroxybenzenesulfonic acid or a salt thereof from acting on a chemical reaction. The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to the embodiment can be used as an inhibitor that inhibits 2,5-dihydroxybenzenesulfonic acid or a salt thereof from acting on an enzyme reaction. The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to the embodiment can be used as an inhibitor that inhibits 2,5-dihydroxybenzenesulfonic acid or a salt thereof from acting on a color reaction using a color-developing reagent. The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to the embodiment can be used as an inhibitor that inhibits 2,5-dihydroxybenzenesulfonic acid or a salt thereof from acting on peroxidase. The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to the embodiment can be used as an inhibitor that inhibits 2,5-dihydroxybenzenesulfonic acid or a salt thereof from acting on hydrogen peroxide.

[0363] Cyclodextrin or its analogues are a collective term for cyclodextrins, which are cyclic oligosaccharides in which several molecules of D-glucose are bound by glycosidic bonds to form a cyclic structure. In cyclodextrin analogues, the hydroxyl groups of D-glucose, which is a component of the cyclodextrin, are substituted. The substituents are not particularly limited. Cyclodextrin analogues are sometimes called chemically modified cyclodextrins. For example, cyclodextrins with a specified degree of substitution, such as methyl-β-cyclodextrin (trade name CAVASOL W7 M, registered trademark) manufactured by Kanto Chemical Co., Ltd., cyclodextrins that are hydrated, and cyclodextrins with a specified purity are available. In the present disclosure, cyclodextrin includes all cyclodextrins regardless of the degree of substitution, the presence or absence of hydration, and the purity. Cyclodextrins are called α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, etc. depending on the size of their ring, i.e., the number of monosaccharides that compose them. For example, when referring to α-cyclodextrin or its analogues, it collectively refers to cyclodextrins or their analogues that have the same ring size as α-cyclodextrin, regardless of whether they are chemically modified or not.

[0364] Any type of cyclodextrin or its analogue may be used as long as it can reduce the effect of 2,5-dihydroxybenzenesulfonic acid or its salt. Examples of cyclodextrin or its analogue include α-cyclodextrin or its analogue, β-cyclodextrin or its analogue, and γ-cyclodextrin or its analogue. The cyclodextrin or its analogue is preferably α-cyclodextrin or its analogue and β-cyclodextrin or its analogue. Depending on the mode of use, β-cyclodextrin or its analogue may be more preferred, or α-cyclodextrin or its analogue may be more preferred.

[0365] Examples of analogs of β-cyclodextrin include methyl-β-cyclodextrin, hydroxyethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, triacetyl-β-cyclodextrin, trimethyl-β-cyclodextrin, dimethyl-β-cyclodextrin, and monomethyl-β-cyclodextrin, some may prefer hydroxypropyl-β-cyclodextrin, and some may prefer methyl-β-cyclodextrin.

[0366] In the inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to the embodiment, the above-mentioned different types of cyclodextrins or analogs thereof may be appropriately mixed.

[0367] The measurement reagent composition according to the present embodiment is a measurement reagent composition configured to measure hydrogen peroxide derived from a substance in a sample, and includes peroxidase, a color-developing reagent configured to react with hydrogen peroxide in the presence of peroxidase to develop color, and an inhibitor that inhibits inhibition of color development by 2,5-dihydroxybenzenesulfonic acid or a salt thereof that may be contained in the sample. As the inhibitor, the above-mentioned cyclodextrin or its analogues can be used.

[0368] The sample may be a biological sample. Examples of biological samples include serum, plasma, blood and urine. Other examples of samples include beverages and foods.

[0369] The concentration of cyclodextrin or its analogue in the reagent composition for measurement is not particularly limited, but the lower limit of the concentration is, for example, 0.1 wt / vol%, preferably 0.5 wt / vol%, and more preferably 1.0 wt / vol%. The upper limit of the concentration is, for example, 35 wt / vol%, preferably 30 wt / vol%, and more preferably 25 wt / vol%.

[0370] The method for measuring a substance in a sample using the measurement reagent composition according to this embodiment is a method for measuring a substance in a sample by an enzymatic method. The method for measuring a substance in a sample is, for example, a method using an oxidase-peroxidase-coloring system, in which a substance to be measured in a sample, which is a specimen, is subjected to an enzymatic reaction to generate hydrogen peroxide, and the hydrogen peroxide is reacted with a coloring reagent in the presence of peroxidase to perform colorimetric quantification. Since the degree of coloring depends on the amount of the substance to be measured, it is possible to quantify the substance to be measured from the degree of coloring. In this disclosure, the amount includes concentration.

[0371] The redox color-developing reaction refers to the Trinder reaction, which develops color by redox reaction, or a reaction using a leuco dye. In the Trinder reaction, for example, a combination of a coupler and a Trinder reagent such as phenol or aniline is used.

[0372] Methods for measuring substances in samples using this principle have already been widely used in the technical field, and the substances to be measured are not particularly limited. The substances to be measured are, for example, biological substances. Examples of the substances to be measured in the measurement reagent composition according to this embodiment include biological components such as uric acid (UA), creatinine (CRE), triglyceride (TG), cholesterol (CHO), and glycated proteins. Examples of glycated proteins include, for example, glycated albumin (GA) and glycated hemoglobin (HbA1c).

[0373] When measuring uric acid, hydrogen peroxide produced by the reaction of uric acid with uricase, an oxidase that uses uric acid as a substrate, can be quantified using a peroxidase-coloring agent system.

[0374] When measuring creatinine, hydrogen peroxide is not directly produced in the reaction between creatinine and creatinine amidinohydrolase, which uses creatinine as a substrate. Therefore, creatine produced in the reaction between creatinine and creatinine amidinohydrolase is further reacted with creatine amide hydrolase contained in the measurement reagent composition to produce sarcosine, and then hydrogen peroxide is produced using sarcosine oxidase (oxidizing enzyme) contained in the measurement reagent composition, thereby performing a so-called coupled reaction, thereby enabling quantification of creatinine using a peroxidase-color developer system.

[0375] When measuring triglyceride, triglyceride can be quantified using a peroxidase-coloring agent system by reacting it with lipoprotein lipase, which uses triglyceride as a substrate, the conjugated enzyme glycerol kinase, and the oxidizing enzyme glycerol-3-phosphate oxidase to produce hydrogen peroxide.

[0376] When measuring cholesterol, cholesterol can be quantified using a peroxidase-coloring agent system by generating free cholesterol using cholesterol esterase, which uses esterified cholesterol in cholesterol as a substrate, and then generating hydrogen peroxide using cholesterol oxidase, an oxidizing enzyme.

[0377] When measuring glycated proteins, first, glycated proteins bound to glucose are decomposed into glycated amino acids or glycated peptides by protease. Next, the glycated amino acids or glycated peptides are decomposed into amino acids or peptides and glucosone by glycated amino acid oxidase. During this process, water and oxygen react with glycated amino acid oxidase to generate hydrogen peroxide, which allows the quantification of glycated proteins using a peroxidase-coloring agent system.

[0378] The measurement reagent composition according to the present embodiment may have various configurations. For example, the reagent composition according to the present embodiment may be a combination of a reagent portion composition configured to be applicable to an automatic analyzer (for example, Hitachi High-Technologies Corporation's automatic analyzer 7180). Here, the reagent portion composition is a composition that constitutes a part of the measurement reagent composition. The reagent portion composition alone cannot measure the substance to be measured. The reagent portion composition may be in a liquid form. The measurement reagent composition according to the present embodiment may be composed of a combination of a dry preparation produced by means of freeze-drying or the like and a dissolving solution. The reagent composition according to the present embodiment may be included in a kit or sensor called a so-called dry system in the form of an enzyme or the like supported on a suitable carrier.

[0379] The measurement reagent composition according to this embodiment may be included in a kit in which the reagent partial composition is divided into two portions (hereinafter, also referred to as a "two-reagent kit"). When using the measurement reagent composition of this embodiment, a first type of reagent partial composition (hereinafter, also referred to as a "first reagent" or "R1") is added to a sample and reacted for a certain period of time, and then a second type of reagent partial composition (hereinafter, also referred to as a "second reagent" or "R2") is further added and reacted, and the change in absorbance during this period is measured, thereby enabling the quantitative determination of the target substance (see, for example, Japanese Patent No. 6446875). The measurement reagent composition according to this embodiment may be included in a kit in which the reagent partial composition is divided into three or more portions (hereinafter, also referred to as a "three-reagent kit"). The measurement reagent composition according to this embodiment may be included in a kit in which the reagent partial composition is divided into a plurality of portions (hereinafter, also referred to as a "multiple-reagent kit").

[0380] When the measurement reagent composition according to this embodiment is included in a kit of a multiple reagent system, cyclodextrin or its analogue may be added to any of the reagent portion compositions. For example, in the case of a two-reagent system kit, cyclodextrin or its analogue may be added to either the first reagent or the second reagent, or both, from the viewpoint of reducing the influence of 2,5-dihydroxybenzenesulfonic acid or its salt on the measured value. In the case of a three-reagent system kit, cyclodextrin or its analogue may be added to either the first reagent, the second reagent, or the third reagent, or a part of these, or all of these, from the viewpoint of reducing the influence of 2,5-dihydroxybenzenesulfonic acid or its salt on the measured value.

[0381] When an automatic analyzer is used, the reaction time and reaction temperature are appropriately determined depending on the type of the automatic analyzer. For example, the automatic analyzer 7180 can perform a reaction for 5 minutes, 10 minutes, 15 minutes, or 22 minutes at 37°C. In the case of a 10-minute reaction, for example, the sample and the first reagent are mixed and reacted at 37°C for 5 minutes, and then the second reagent is added, mixed, and reacted for another 5 minutes.

[0382] The type of automatic analyzer that can be used is not particularly limited, but is preferably an automatic analyzer for biochemistry or an automatic analyzer for biochemistry and immunocomplex. For example, the automatic analyzer 7180, the automatic analyzer 3500, the automatic analyzer 3100, the automatic analyzer LABOSPECT008α, the automatic analyzer LABOSPECT006, the automatic analyzer LABOSPECT003 (all manufactured by Hitachi High-Technologies Corporation), TBA (registered trademark)-FX8, TBA (registered trademark)-c16000, TBA (registered trademark)-2000FR, TBA (registered trademark)-1500FR, TBA (registered trademark)-nx360, TBA (registered trademark)-c8000, TBA (registered trademark)-120FR Pearl Edition, TBA (registered trademark)-120FR Sora Edition, TBA (registered trademark)-c4000, Accute (registered trademark)-RX (all manufactured by Canon Medical Systems, Inc.), JCA-ZS050 automatic analyzer BioMajesty (registered trademark) ZERO, JCA-BM6010 G automatic analyzer BioMajesty(registered trademark), JCA-BM6050 Automated Analyzer BioMajesty(registered trademark), JCA-BM9130 Automated Analyzer BioMajesty(registered trademark), JCA-BM6070 Automated Analyzer BioMajesty(registered trademark), JCA-BM8000 series Automated Analyzer BioMajesty(registered trademark) (all manufactured by JEOL Ltd.), DxC 700 AU, AU5800, AU680, AU480 (all manufactured by Beckman Coulter, Inc.), VITROS(registered trademark) XT 7600, VITROS(registered trademark) 5600 II, VITROS(registered trademark) 4600, VITROS(registered trademark) 350PLUS (all manufactured by Ortho Diagnostics, Inc.), cobas 8000 <702> , cobas 8000 <502> , cobas 6000 <501> (All manufactured by Roche Diagnostics), Atellica CH 930, Dimension EXL 200, Dimension EXL LM, Dimension Vista 500T / 1500T, Dimension Vista 1000T / 3000T (All manufactured by Siemens Healthcare), etc.

[0383] As the peroxidase, any type of enzyme may be used as long as it catalyzes the reaction between hydrogen peroxide and a redox color-developing reagent, and there are no particular limitations on the amount used or the form of addition.

[0384] Peroxidase of any origin can be used, and examples thereof include peroxidase derived from plants such as horseradish, and peroxidase derived from microorganisms such as bacteria and mold. Specific examples include Peroxidase from horseradish (Sigma), peroxidase from horseradish (Wako Pure Chemical Industries, Ltd.), PO "AMANO" 3 (Amano Enzyme Co., Ltd.), and Peroxidase (Toyobo Enzyme Co., Ltd.). In addition, if there is no suitable enzyme that catalyzes the reaction of directly oxidizing the substance to be measured to generate hydrogen peroxide, it is also possible to measure the amount of the substance to be measured by appropriately designing a conjugated reaction that combines an enzyme (several stages of enzyme reactions may be connected) that catalyzes a reaction that can change the substance to be measured into a substrate that can generate hydrogen peroxide and an oxidase.

[0385] The concentration of peroxidase in the measurement reagent composition may be sufficient to react with the generated hydrogen peroxide, and the lower limit is, for example, 0.01 U / mL or more, preferably 0.1 U / mL or more, and more preferably 1 U / mL or more. From the viewpoint of cost, the upper limit is, for example, 500 U / mL or less, preferably 100 U / mL or less, and more preferably 50 U / mL or less.

[0386] Examples of Trinder reagents as dye precursors used in the Trinder reaction, which is an oxidation-reduction color-developing reaction, include phenol, phenol derivatives, aniline derivatives, naphthol, naphthol derivatives, naphthylamine, and naphthylamine derivatives. For example, N-(3-sulfopropyl)aniline sodium monohydrate (HALPS), N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium monohydrate (TOPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (MAOS), N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (HDAPS), N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (HDAOS), N-ethyl-N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (DAPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium (DAOS), N-ethyl-N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (DAPS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (DAOS), N-ethyl-N-(3-sulfopropyl)-3,5-dimethoxyaniline sodium monohydrate (DAPS), N-ethyl-N-( ... Examples of suitable aniline salts include sodium N-ethyl-N-(3-sulfopropyl)aniline (ALPS), sodium N-ethyl-N-(3-sulfopropyl)-3-methoxyaniline monohydrate (ADPS), sodium N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methoxyaniline dihydrate (ADOS), N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine (TOOS), disodium N,N-bis(4-sulfobutyl)-3-methylaniline (TODB), and 3-hydroxy-2,4,6-triiodobenzoic acid (HTIB). Of these, HALPS, TOPS, MAOS, HDAPS, HDAOS, DAPS, DAOS, ALPS, ADPS, ADOS, TOOS, TODB, or HTIB is preferred, and TODB is more preferred.

[0387] The color-developing wavelength when TOPS is used is 550 nm. The color-developing wavelength when ALPS is used is 561 nm. The color-developing wavelength when TOOS is used is 555 nm. The color-developing wavelength when TODB is used is 550 nm. However, the wavelength measured by the device for measuring absorbance does not have to strictly match the color-developing wavelength. For example, it is possible to measure the color-developing reactions when TOPS, ALPS, TOOS, and TODB are used with a device that measures absorbance at a wavelength of 546 nm.

[0388] In addition, if the Trinder reagent and the coupler are contained in the same reagent composition, the Trinder reagent and the coupler will naturally undergo an oxidative condensation reaction and will develop a color during storage, so it is preferable that the Trinder reagent and the coupler are packaged separately in separate reagent partial compositions. A partial composition of the reagent composition containing the Trinder reagent is called a Trinder reagent-containing partial composition. Also, a partial composition of the reagent composition containing the coupler is called a coupler-containing partial composition.

[0389] The concentration of the Trinder's reagent contained in the Trinder's reagent-containing partial composition may be sufficient to react with the generated hydrogen peroxide, and examples of the lower limit include 0.1 mmol / L or more, preferably 0.2 mmol / L or more, more preferably 0.5 mmol / L or more, and even more preferably 1.0 mmol / L or more. Examples of the upper limit include 50 mmol / L or less, preferably 30 mmol / L or less, more preferably 20 mmol / L or less, and even more preferably 10 mmol / L or less.

[0390] Couplers that undergo an oxidative condensation reaction with the Trinder reagent include 4-aminoantipyrine (4AA), aminoantipyrine derivatives, vanillindiaminesulfonic acid, methylbenzthiazolinone hydrazone (MBTH), and sulfonated methylbenzthiazolinone hydrazone (SMBTH).

[0391] The concentration of the coupler contained in the coupler-containing partial composition may be sufficient to react with the generated hydrogen peroxide, and examples of the lower limit include 0.1 mmol / L or more, preferably 0.2 mmol / L or more, more preferably 0.5 mmol / L or more, and more preferably 1.0 mmol / L or more. Examples of the upper limit include 50 mmol / L or less, preferably 30 mmol / L or less, preferably 20 mmol / L or less, and more preferably 10 mmol / L or less.

[0392] Examples of leuco dyes include 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 10-(carboxymethylaminocarbonyl)-3,7-bis(dimethylamino)phenothiazine sodium (DA-67), N-(carboxymethylaminocarbonyl)-4,4-bis(dimethylamino)biphenylamine (DA-64), and N,N,N',N',N'',N''-hexa(3-sulfopropyl)-4,4',4''-triaminotriphenylmethane 6Na (TPM-PS), of which DA-64 or DA-67 is preferred, and DA-67 is more preferred.

[0393] The concentration of the leuco dye in the measurement reagent composition may be sufficient to react with the generated hydrogen peroxide, and examples of the lower limit include 0.005 mmol / L or more, preferably 0.01 mmol / L or more, more preferably 0.02 mmol / L or more, and even more preferably 0.05 mmol / L or more. Examples of the upper limit include 10 mmol / L or less, preferably 5 mmol / L or less, preferably 3 mmol / L or less, and even more preferably 1 mmol / L or less.

[0394] The measuring reagent composition may further include a ferrocyanide. This may further enhance the effect of cyclodextrin or its analogues. Examples of the ferrocyanide include any compound containing ferrocyanide ions, such as potassium ferrocyanide (Fe(CN) 6 K 4) and sodium ferrocyanide (Fe(CN) 6 Na 4 ) etc.

[0395] The concentration of the ferrocyanide in the measuring reagent composition is not particularly limited, but examples of the lower limit include 0.001 mmol / L or more, preferably 0.005 mmol / L or more, and more preferably 0.01 mmol / L or more. Examples of the upper limit include 1.0 mmol / L or less, preferably 0.5 mmol / L or less, and more preferably 0.1 mmol / L or less.

[0396] The reagent composition for measurement may further contain a saccharide other than cyclodextrin or its analogue, which may further enhance the effect of cyclodextrin or its analogue.

[0397] Examples of sugars include sugar alcohols other than cyclodextrin, aldoses, ketoses, non-reducing disaccharides, and reducing disaccharides. Examples of sugar alcohols other than cyclodextrin include erythritol, xylitol, sorbitol, and mannitol. Examples of aldoses include erythrose, xylose, glucose, and mannose. Examples of ketoses include erythrulose, xylulose, and fructose. Examples of non-reducing disaccharides include trehalose and sucrose. Examples of reducing disaccharides include maltose, and sorbitol is preferred.

[0398] The concentration of the saccharide in the reagent composition for measurement is not particularly limited, but the lower limit is, for example, 0.1 wt / vol % or more, preferably 0.5 wt / vol % or more, and more preferably 1.0 wt / vol % or more, and the upper limit is, for example, 50 wt / vol % or less, preferably 40 wt / vol % or less, and more preferably 30 wt / vol % or less.

[0399] The measurement reagent composition for measuring glycated proteins may further include a protease. When measuring glycated proteins, the protease contained in the reagent composition may be a protease that effectively generates glycated amino acids or glycated peptides by decomposing glycated proteins derived from human proteins. The protease is, for example, an endopeptidase. An example of the endopeptidase is serine endopeptidase. The protease is preferably a protease derived from a microorganism such as the genus Bacillus, the genus Streptomyces, the genus Tritirachium, or the genus Aspergillus.

[0400] Examples of proteases derived from the genus Bacillus include Alcalase, Neutrase, Esperase, and Savinase (all manufactured by Novozymes), Bioprase OP, Bioprase SP-20FG, Bioprase 30L, Bioprase 30G, Bioprase AL-15FG, Bioprase APL-30, and Protease CL-15 (all manufactured by Nagase ChemteX Corporation), Protin SD PC-10CF, Samoase PC-10F, Protin SD-AY10, and Protin SD-NY10 (all manufactured by Amano Enzyme Co., Ltd.), Multifect PR6L, Optimase PR40L, Optimase PR40X, and Optimase PR40E (all manufactured by Danisco Japan Co., Ltd.), Orientase 22BF, Nucleisin, Orientase 10NL, and Orientase 90N (all manufactured by HI), and Aroase. Examples of such protease include Aroase AP-10, Aroase NP-10, Aroase XA-10 (all manufactured by Yakult Pharmaceutical Co., Ltd.), subtilisin, bacterial proteinase type XXIV, protease type VIII derived from Bacillus licheniformis, protease type IX derived from Bacillus polymyxa, thermolysin type X derived from Geobacillus Stearothermophilus, protease type XV derived from Bacillus polymyxa, bacterial proteinase type XXIV, protease type XXVII derived from Bacillus sp., protease type XXXI derived from Bacillus licheniformis (all manufactured by Sigma), thermolysin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), Dispase I, Dispase II (all manufactured by Godo Shusei Co., Ltd.), and Neutral Proteinase (manufactured by Toyobo USA, Inc.).

[0401] Examples of proteases derived from the genus Streptomyces include pronase, protease Streptomyces griseus type XIV (all manufactured by Sigma), Alkalophilic proteinase (manufactured by Toyobo USA), and Denatiyme PMC SOFTER (manufactured by Nagase ChemteX Corporation).

[0402] An example of a protease derived from the genus Tritirachium is Proteinase K (Sigma).

[0403] Examples of proteases derived from the genus Aspergillus include Protease P "Amano" 3SD, Protease A "Amano" SD, Protease M "Amano" SD (all manufactured by Amano Enzyme Co., Ltd.), Sumiteam MP, Sumiteam LPL-G, Sumiteam LP50D, Sumiteam AP (all manufactured by Shin Nippon Chemical Industry Co., Ltd.), Orientase OP, Orientase AY (all manufactured by HBI Corporation), Protease Type XIII derived from Aspergillus saitoi, Protease Type XIX derived from Aspergillus sojae, Protease Type XXIII derived from Aspergillus melleus, Protease Type XXIII derived from Aspergillus oryzae (all manufactured by Sigma), Denatzyme AP, Denapsin 2P (all manufactured by Nagase ChemteX Corporation), Panchidase NP-2, Panchidase MP, and Panchidase P (both manufactured by Yakult Pharmaceutical Co., Ltd.).

[0404] Of these, Alcalase, Bioprase SP-20FG, Protin SD-AY10, Multifect PR6L, Optimase PR40L, Aroase XA-10, Subtilisin, Pronase, Proteinase K, and PR "Amano" K are preferred.

[0405] From another viewpoint, preferred examples of the protease contained in the 4-aminoantipyrine-containing partial composition of the reagent composition for measuring a glycosylated protein according to the embodiment include proteases having an enzyme number of EC: 3.4 or EC: 3.4.21. Further preferred examples include proteases having an enzyme number of EC: 3.4.21.62.

[0406] The concentration of the protease in the measurement reagent composition may be any concentration that can decompose the glycated protein within a suitable time, and examples of the lower limit include 100 U / mL or more, preferably 1 kU / mL or more, and more preferably 10 kU / mL or more. Examples of the upper limit include 1000 kU / mL or less, preferably 200 kU / mL or less, and more preferably 100 kU / mL or less, from the viewpoint of cost.

[0407] The measurement reagent composition for measuring glycated proteins may further comprise a glycated amino acid oxidase. When measuring glycated proteins, the glycated amino acid oxidase contained in the reagent composition may be any glycated amino acid oxidase that effectively acts on glycated amino acids or glycated peptides derived from human proteins, and examples of such glycated amino acid oxidase include those derived from the genus Gibberella, the genus Aspergillus, the genus Candida (C. andida, Penicillium, Fusarium, Acremonium, Deb aryomyces and Corynebacterium, and recombinant glycosylated amino acid oxidase mutants thereof. More specifically, ketoamine oxidase (KAOD, manufactured by Asahi Kasei Pharma Corporation, described in Clinica Chimica Acta, 2002, 324, pp. 61-71), mutant KAOD (KAOD-V, manufactured by Asahi Kasei Pharma Corporation, described in Patent Document 1), and FAOD-E (manufactured by Kikkoman Corporation) are included.

[0408] The concentration of glycated amino acid oxidase in the measurement reagent composition may be sufficient to react with the generated glycated amino acid or glycated peptide, and examples of the lower limit include 0.1 U / mL or more, preferably 1 U / mL or more, and more preferably 10 U / mL or more. From the viewpoint of cost, examples of the upper limit include 1000 U / mL or less, preferably 200 U / mL or less, and more preferably 100 U / mL or less.

[0409] The measurement reagent composition may further contain, as a solvent, water and an organic solvent, etc. Examples of the organic solvent include alcohol-based solvents (alcohols having 1 to 18 carbon atoms, specifically, methanol, butanol, ethylene glycol, glycerin, etc.), ketone-based solvents (acetone, methyl ethyl ketone, etc.), and ether-based solvents (tetrahydrofuran, diethyl ether, ethylene glycol monoalkyl ether, cyclic ether, etc.).

[0410] The measurement reagent composition may also contain a buffer. Examples of the buffer include N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid (TES), 3-[4-(2-hydroxyethyl)-1-piperazinyl]propanesulfonic acid (EPPS), 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid (HEPES), 2-hydroxy-3-[4-(2-hydroxyethyl)-1-piperazinyl]propanesulfonic acid (HEPPSO), N-(2-acetamido)-2-aminoethanesulfonic acid (ACES), N-(2-acetamido)-1-aminoethanesulfonic acid (N-(2-acetamido)-1-aminoethanesulfonic acid ... Aminodiacetic acid (ADA), N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES), N,N-bis(2-hydroxyethyl)glycine (Bicine), Bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane (Bis-Tris), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), N-cyclohexyl-2-hydroxy-3-aminopropanesulfonic acid (CAPSO), N-cyclohexyl-2-aminoethanesulfonic acid (CHES), 3-[N,N-Bis(2-hydroxyethyl)glycine (Bicine), N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS), N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS), N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS), N-tris(hydroxyethyl ... Examples of buffers include those using 2-hydroxy-N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPSO), N-[tris(hydroxymethyl)methyl]glycine (Tricine), and trishydroxymethylaminomethane (Tris), as well as boric acid, ammonia, glycine, carbonic acid, acetic acid, phosphoric acid, diethanolamine, p-phenolsulfonic acid, 2-amino-2-methylpropane-1,3-diol, cacodylic acid, maleic acid, veronal, and 3,3-dimethylglutaric acid.

[0411] The pH of the reagent composition using a buffer can be appropriately determined taking into consideration the optimum pH and stability of the enzyme used, the stability of the color-developing reagent used, and the like.

[0412] The measurement reagent composition may further contain other additives such as preservatives, enzyme stabilizers, surfactants, and other enzymes. Examples of preservatives include sodium azide and ProClin. Any enzyme stabilizer that stabilizes the enzyme during storage can be used. In addition, the above-mentioned sugars such as cyclodextrin, sucrose, mannose, fructose, lactose, galactose, and trehalose, and sugar alcohols such as sorbitol and mannitol, which reduce the effects of calcium dobesilate, can also function as enzyme stabilizers.

[0413] The surfactant may be any of cationic surfactants, anionic surfactants, amphoteric surfactants, and nonionic surfactants. Examples of nonionic surfactants include polyoxyethylene alkyl ethers, polyoxyethylene alkylphenyl ethers, polyoxyethylene polyoxypropylene alkyl ethers, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters, glycerin fatty acid esters, and polyoxyethylene polyoxypropylene block polymers. Examples of amphoteric surfactants include alkyl betaines, betaine acetates, sulfobetaines, and alkylamine oxides.

[0414] Other enzymes include, for example, ascorbic acid oxidase and bilirubin oxidase, which are used to suppress the influence of ascorbic acid and bilirubin contained in the sample on the measured value.

[0415] As a method for confirming the effect of the measurement reagent composition according to this embodiment in suppressing the influence of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on the measured value, the measurement value obtained by measuring a sample containing 2,5-dihydroxybenzenesulfonic acid or a salt thereof using the measurement reagent composition according to this embodiment is B, and the measurement value obtained by measuring a sample not containing 2,5-dihydroxybenzenesulfonic acid or a salt thereof using the measurement reagent composition according to this embodiment is C, and the accuracy A (%) is calculated using the following formula (1). The closer the accuracy A is to 100%, the more the measurement reagent composition according to this embodiment exerts the effect of suppressing the influence of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on the measured value. A = (B / C) × 100 (1)

[0416] Examples of accuracy A include 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more, 97% or more, 98% or more, or 99% or more. EXAMPLES

[0417] Next, examples of the present embodiment will be described, but it goes without saying that the present embodiment is not limited to the following examples.

[0418] For the NMR measurements in Examples 1 and 2, AVANCE3HD400MHz (manufactured by Bruker) was used.

[0419] A Hitachi 7180 automatic analyzer (Hitachi High-Technologies Corporation) was used to measure absorbance in Examples 3 to 10. In Examples 3 and 5 to 9, which used the Trinder reagent and coupler, the main wavelength was set to 546 nm and the secondary wavelength was set to 700 nm in the analyzer. The absorbance Abs of the sample was calculated by dividing the absorbance at a wavelength of 546 nm by Abs. 546 The absorbance at a wavelength of 700 nm is Abs 700 This was calculated using the following formula (2). Abs=Abs 546 -Abs 700(2)

[0420] In the measurement of absorbance in Example 9, the main wavelength was set to 600 nm and the sub-wavelength was set to 750 nm in the device. The absorbance Abs of the sample was calculated by dividing the absorbance at a wavelength of 600 nm by Abs 600 The absorbance at a wavelength of 750 nm is Abs 750 This was calculated using the following formula (3). Abs=Abs 600 -Abs 750 (3)

[0421] In the measurement of absorbance in Example 10, the main wavelength was set to 600 nm and the sub-wavelength was set to 700 nm in the device. The absorbance Abs of the sample was calculated by Abs 600 The absorbance at a wavelength of 700 nm is Abs 700 was calculated using the following formula (4). Abs=Abs 600 -Abs 700 (4)

[0422] In Example 4, in which a leuco dye was used, the main wavelength was set to 660 nm and the sub-wavelength was set to 800 nm in the device. The absorbance Abs of the sample was calculated by Abs 660 The absorbance at a wavelength of 800 nm is Abs 800 This was calculated using the following formula (5). Abs=Abs 660 -Abs 800 (5)

[0423] [Example 1: Analysis of the interaction between cyclodextrin and 2,5-dihydroxybenzenesulfonate by one-dimensional NMR]

[0424] (liquid preparation) As mother liquid 1 for sample preparation, calcium dobesilate (Tokyo Chemical Industry Co., Ltd.) was dissolved in heavy water (Taiyo Nissan Co., Ltd.) to prepare a 0.024 mol / L calcium dobesilate solution. As mother liquid 2 for sample preparation, methyl-β-cyclodextrin (Aldrich Co., Ltd.) was dissolved in heavy water (Taiyo Nissan Co., Ltd.) to prepare a 0.024 mol / L methyl-β-cyclodextrin solution. By appropriately mixing mother liquid 2 and heavy water (Taiyo Nissan Co., Ltd.) with 70 μL of mother liquid 1, samples with the substance ratios of calcium dobesilate and methyl-β-cyclodextrin of 1:0.25, 1:0.5, 1:1, 1:2, 1:3, and 1:4 were obtained. In addition, 2,5-dihydroxybenzenesulfonic acid was considered as one unit of calcium dobesilate, and the molecular weight of methyl-β-cyclodextrin was set to 1310.

[0425] (measurement) One-dimensional NMR measurements were performed to obtain spectra for each sample. The results are shown in Figure 1. As shown in Figure 1, as the ratio of methyl-β-cyclodextrin to calcium dobesilate increased, a peak shift was observed in the spectrum. This indicated that methyl-β-cyclodextrin and calcium dobesilate interacted with each other. It was also shown that the interaction was dependent on the concentration of cyclodextrin.

[0426] [Example 2: Analysis of the interaction between cyclodextrin and 2,5-dihydroxybenzenesulfonate by 2D NMR]

[0427] (liquid preparation) As the mother liquid 1 for sample preparation, 5 mg of calcium dobesilate (Tokyo Chemical Industry Co., Ltd.) and 125 mg of methyl-β-cyclodextrin (Aldrich Co., Ltd.) were dissolved in 700 μL of heavy water (Taiyo Nissan Co., Ltd.) to obtain a sample with a substance ratio of calcium dobesilate to methyl-β-cyclodextrin of 1:4. Note that, for calcium dobesilate, 2,5-dihydroxybenzenesulfonic acid was considered as one unit, and the molecular weight of methyl-β-cyclodextrin was set to 1310.

[0428] (measurement) Two-dimensional NMR measurements were performed to obtain NOESY (Nuclear Overhauser Enhancement Spectroscopy) spectra for the samples. The results are shown in Figure 2. As shown in Figure 2, a nuclear Overhauser effect (NOE) correlation was observed between methyl-β-cyclodextrin and calcium dobesilate, indicating that methyl-β-cyclodextrin and calcium dobesilate interact with each other.

[0429] [Example 3: Effect of adding cyclodextrin or its analogue and ferrocyanide to a measuring reagent composition using Trinder reagent and coupler to avoid the influence of 2,5-dihydroxybenzenesulfonic acid]

[0430] (First Reagent) The following reagent components were dissolved in purified water to the concentrations shown below, and the pH was adjusted to 7.5 to prepare a first reagent. In addition, a control reagent was prepared that did not contain cyclodextrin or its analogues. The cyclodextrin or its analogues used in Example 3 were α-cyclodextrin (manufactured by Wako Pure Chemical Industries, Ltd.), methyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), hydroxypropyl-β-cyclodextrin (manufactured by Aldrich Co., Ltd.), γ-cyclodextrin (manufactured by Wako Pure Chemical Industries, Ltd.), trimethyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), and hydroxyethyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.). 100mmol / L N-tris(hydroxymethyl)aminomethane (Dojindo Chemical Industries, Ltd.) 1% w / v or 5% w / v cyclodextrin or analogue

[0431] (Second Reagent) The following reagent components were dissolved in purified water to the concentrations shown below, and the pH was adjusted to 7.5 to prepare a second reagent. A control reagent was also prepared that did not contain potassium ferrocyanide. 10U / mL peroxidase (Amano Enzyme Co., Ltd.) 100mmol / L N-tris(hydroxymethyl)aminomethane (Dojindo Chemical Industries, Ltd.) 5mmol / L 4-aminoantipyrine (Actec) 5mmol / L N,N-bis(4-sulfobutyl)-3-methylaniline disodium (Dojin Chemical Industries, Ltd.) 0.1mmol / L Potassium ferrocyanide (Kokusan Chemical Co., Ltd.)

[0432] (Sample specimen) As the mother solution for preparing the sample solution, a hydrogen peroxide solution (manufactured by Kokusan Kagaku Co., Ltd.) was diluted with purified water to prepare a 3.5 mmol / L hydrogen peroxide solution. As the additive solution for preparing the sample solution, calcium dobesilate (manufactured by Tokyo Kasei Co., Ltd.) was diluted with purified water to prepare 640 μg / mL, 1000 μg / mL, and 2000 μg / mL calcium dobesilate solutions. The mother solution was added to each additive solution or purified water at a ratio of 9:1, and then mixed appropriately to obtain hydrogen peroxide solutions containing calcium dobesilate at concentrations of 0 μg / mL, 16 μg / mL, 32 μg / mL, 64 μg / mL, 100 μg / mL, and 200 μg / mL, respectively.

[0433] (measurement) 4.5 μL of the sample specimen was added to 180 μL of the first reagent incubated at 37° C., and allowed to react for 5 minutes at 37° C. Next, 45 μL of the second reagent was added to the mixture of the first reagent and the sample specimen, and allowed to react for an additional 5 minutes at 37° C. The absorbance Abs1 5 minutes after the sample specimen was added to the first reagent, and the absorbance Abs2 5 minutes after the second reagent was added were measured, and the absorbance change V was calculated according to the following formula (6). V = Abs2 - Abs1 × 180 / 225 (6) Here, 180 represents the liquid volume of the first reagent, and 225 represents the total liquid volume of the first and second reagents.

[0434] The absorbance change measured using purified water instead of the sample specimen is used as the reagent blank, and the absorbance change V measured using the sample specimen is expressed as shown in the following formula (7). S Change in absorbance of the reagent blank from V B The value obtained by subtracting this was calculated as the sensitivity ΔAbs. ΔAbs=V S -V B (7)

[0435] As shown in the following formula (8), the sensitivity C measured using a sample specimen not containing calcium dobesilate was set to 100%, and the percentage of sensitivity B measured using a sample specimen containing calcium dobesilate was calculated as accuracy A. A = (B / C) × 100 (8) The results are shown in Tables 1 and 2. [Table 1-1] [Table 1-2] [Table 2-1] [Table 2-2]

[0436] As shown in Table 1, when a reagent without cyclodextrin or its analogue was used, the accuracy of the measurement was adversely affected by the addition of calcium dobesilate in a concentration-dependent manner. In contrast, the effect of calcium dobesilate on the measurement was reduced by adding cyclodextrin or its analogue to the reagent. For example, the addition of α-cyclodextrin, methyl-β-cyclodextrin, γ-cyclodextrin, trimethyl-β-cyclodextrin, or hydroxyethyl-β-cyclodextrin to the reagent improved the accuracy of the measurement depending on the concentration of cyclodextrin or its analogue. Therefore, it was shown that in a measurement system using a redox color-developing reagent, the effect of calcium dobesilate contained in the sample on the measurement can be reduced by including cyclodextrin or its analogue in the reagent.

[0437] As shown in Table 2, even when the reagent did not contain potassium ferrocyanide, the accuracy of the measurement was adversely affected by the addition of calcium dobesilate in a concentration-dependent manner when a reagent to which cyclodextrin or its analogues had not been added was used. In contrast, the effect of calcium dobesilate on the measurement was reduced by adding cyclodextrin or its analogues to the reagent. For example, the addition of α-cyclodextrin, methyl-β-cyclodextrin, γ-cyclodextrin, trimethyl-β-cyclodextrin, or hydroxyethyl-β-cyclodextrin to the reagent improved the accuracy of the measurement depending on the concentration of cyclodextrin or its analogues.

[0438] As shown by comparing Tables 1 and 2, the effect on calcium dobesilate measurement can be reduced by adding potassium ferrocyanide to the reagent. For example, even if cyclodextrin or its analogue is not added to the reagent, the effect on calcium dobesilate measurement can be reduced by adding potassium ferrocyanide to the reagent in the entire calcium dobesilate concentration range tested. Also, even when 1% α-cyclodextrin, 1% or 5% methyl-β-cyclodextrin, 1% or 5% hydroxypropyl-β-cyclodextrin, 1% or 5% γ-cyclodextrin, 1% or 5% trimethyl-β-cyclodextrin, or 1% or 5% hydroxyethyl-β-cyclodextrin is added to the reagent, the effect on calcium dobesilate measurement can be reduced by adding potassium ferrocyanide to the reagent in the entire calcium dobesilate concentration range tested. When 5% α-cyclodextrin was added to the reagent, the effect on calcium dobesilate measurement was reduced when potassium ferrocyanide was added to the reagent at calcium dobesilate concentrations of 64 to 200 μg / mL.

[0439] Therefore, it was shown that in a measurement system using an oxidation-reduction color-developing reagent, the effect of adding potassium ferrocyanide to the measurement reagent composition can also be reduced on the measurement of calcium dobesilate contained in the sample.

[0440] [Example 4: Effect of adding cyclodextrin or its analogue and ferrocyanide to a measuring reagent composition using a leuco dye to avoid the influence of 2,5-dihydroxybenzenesulfonic acid]

[0441] (First Reagent) The following reagent components were dissolved in purified water to the concentrations shown below, and the pH was adjusted to 7.5 to prepare a first reagent. In addition, a control reagent was prepared that did not contain cyclodextrin or its analogues. The cyclodextrin or its analogues used in Example 4 were α-cyclodextrin (manufactured by Wako Pure Chemical Industries, Ltd.), methyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), hydroxypropyl-β-cyclodextrin (manufactured by Aldrich Co., Ltd.), γ-cyclodextrin (manufactured by Wako Pure Chemical Industries, Ltd.), trimethyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), and hydroxyethyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.). 100mmol / L N-tris(hydroxymethyl)aminomethane (Dojindo Chemical Industries, Ltd.) 1%, 3%, or 5% w / v cyclodextrin or its analogues

[0442] (Second Reagent) The following reagent components were dissolved in purified water to the concentrations shown below, and the pH was adjusted to 7.5 to prepare a second reagent. A control reagent was also prepared that did not contain potassium ferrocyanide. 10U / mL peroxidase (Amano Enzyme Co., Ltd.) 100mmol / L N-tris(hydroxymethyl)aminomethane (Dojindo Chemical Industries, Ltd.) 2mmol / L DA-67 (Fujifilm Wako Pure Chemical Industries, Ltd.) 0.1mmol / L Potassium ferrocyanide (Kokusan Chemical Co., Ltd.)

[0443] (Sample specimen) As the mother liquor for preparing the sample solution, a hydrogen peroxide solution (manufactured by Kokusan Kagaku Co., Ltd.) was diluted with purified water to prepare a 3.5 mmol / L hydrogen peroxide solution. As the additive solution for preparing the sample solution, calcium dobesilate (manufactured by Tokyo Kasei Co., Ltd.) was diluted with purified water to prepare 640 μg / mL, 1000 μg / mL, and 2000 μg / mL calcium dobesilate solutions. The mother liquor was mixed with each additive solution or purified water at a ratio of 9:1, and then mixed appropriately to obtain hydrogen peroxide solutions containing calcium dobesilate at concentrations of 0 μg / mL, 16 μg / mL, 32 μg / mL, 64 μg / mL, 100 μg / mL, and 200 μg / mL.

[0444] (measurement) 4.5 μL of the sample specimen was added to 180 μL of the first reagent incubated at 37° C., and allowed to react for 5 minutes at 37° C. Next, 45 μL of the second reagent was added to the mixture of the first reagent and the sample specimen, and allowed to react for an additional 5 minutes at 37° C. The absorbance Abs1 5 minutes after the sample specimen was added to the first reagent, and the absorbance Abs2 5 minutes after the second reagent was added were measured, and the absorbance change V was calculated according to the following formula (9). V=Abs2-Abs1×180 / 225 (9) Here, 180 represents the liquid volume of the first reagent, and 225 represents the total liquid volume of the first and second reagents.

[0445] The absorbance change measured using purified water instead of the sample specimen is used as the reagent blank, and the absorbance change V measured using the sample specimen is calculated as shown in the following formula (10). S Change in absorbance of the reagent blank from V B The value obtained by subtracting this was calculated as the sensitivity ΔAbs. ΔAbs=V S -V B (10)

[0446] As shown in the following formula (11), the sensitivity C measured using a sample specimen not containing calcium dobesilate was set as 100%, and the percentage of sensitivity B measured using a sample specimen containing calcium dobesilate was calculated as accuracy A. A = (B / C) × 100 (11) The results are shown in Tables 3 and 4.

[0447] [Table 3-1] [Table 3-2] [Table 4-1] [Table 4-2]

[0448] As shown in Table 3, when a reagent without cyclodextrin or its analogue was used, the accuracy of the measurement was adversely affected by the addition of calcium dobesilate in a concentration-dependent manner. In contrast, the effect of calcium dobesilate on the measurement was reduced by adding cyclodextrin or its analogue to the reagent. For example, the addition of α-cyclodextrin, methyl-β-cyclodextrin, γ-cyclodextrin, trimethyl-β-cyclodextrin, or hydroxyethyl-β-cyclodextrin to the reagent improved the accuracy of the measurement depending on the concentration of cyclodextrin or its analogue. Therefore, it was shown that in a measurement system using a redox color-developing reagent, the effect of calcium dobesilate contained in the sample on the measurement can be reduced by including cyclodextrin or its analogue in the reagent.

[0449] As shown in Table 4, even when the reagent did not contain potassium ferrocyanide, the accuracy of the measurement was adversely affected by the addition of calcium dobesilate in a concentration-dependent manner when a reagent was used that did not contain cyclodextrin or its analogues. In contrast, the effect of calcium dobesilate on the measurement was reduced by adding cyclodextrin or its analogues to the reagent. For example, the addition of 1% or 5% α-cyclodextrin or 1% γ-cyclodextrin to the reagent improved the accuracy of the measurement depending on the concentration of cyclodextrin or its analogues.

[0450] As shown by the comparison of Tables 3 and 4, the effect on the measurement of calcium dobesilate can be reduced by adding potassium ferrocyanide to the reagent. For example, even if cyclodextrin or its analogue is not added to the reagent, the effect on the measurement of calcium dobesilate can be reduced by adding potassium ferrocyanide to the reagent in the entire calcium dobesilate concentration range tested. Also, even when 1% or 5% α-cyclodextrin, 1% or 5% methyl-β-cyclodextrin, 1% or 5% hydroxypropyl-β-cyclodextrin, 1% or 5% γ-cyclodextrin, 1% or 3% trimethyl-β-cyclodextrin, or 1% or 5% hydroxyethyl-β-cyclodextrin is added to the reagent, the effect on the measurement of calcium dobesilate can be reduced by adding potassium ferrocyanide to the reagent in the entire calcium dobesilate concentration range tested.

[0451] Therefore, it was shown that in a measurement system using an oxidation-reduction color-developing reagent, the effect of adding potassium ferrocyanide to the measurement reagent composition can also be reduced on the measurement of calcium dobesilate contained in the sample.

[0452] [Example 5: Effects of the type and concentration of cyclodextrin in a glycated albumin measurement reagent]

[0453] (First Reagent) The following reagent components were dissolved in purified water to the concentrations shown below, and the pH was adjusted to 7.6 to prepare a first reagent. In addition, a control reagent was prepared that did not contain cyclodextrin or its analogues. The cyclodextrin or its analogues used in Example 5 were α-cyclodextrin (manufactured by Wako Pure Chemical Industries, Ltd.), methyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), hydroxypropyl-β-cyclodextrin (manufactured by Aldrich Co., Ltd.), and γ-cyclodextrin (manufactured by Wako Pure Chemical Industries, Ltd.). 100mmol / L N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid (Dojin Chemical Industries, Ltd.) 0.05% by weight / volume sodium azide (Merck) 1mmol / L Sodium ethylenediaminetetraacetate (Kokusan Chemical Co., Ltd.) 6% by weight / volume D(-)-sorbitol (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) 2mmol / L N,N-bis(4-sulfobutyl)-3-methylaniline disodium (Dojin Chemical Industries, Ltd.) 10U / mL peroxidase (Amano Enzyme Co., Ltd.) 30U / mL ketoamine oxidase (Asahi Kasei Pharma) 1%, 3%, or 5% w / v cyclodextrin or its analogues

[0454] (Second Reagent) Lucica GA-L GA R-2 (Asahi Kasei Pharma Corporation), which contains protease and 4-AA, was used.

[0455] (Sample specimen) As the mother liquor for preparing the samples, Lucica GA-L Dedicated Control Serum L (Asahi Kasei Pharma) was prepared. As the additive liquid for preparing the samples, calcium dobesilate (Tokyo Kasei) was diluted with purified water to prepare a 640μg / mL calcium dobesilate aqueous solution. The mother liquor was mixed with each additive liquid or purified water at a ratio of 9:1, and then mixed appropriately to obtain sample samples containing calcium dobesilate at concentrations of 0μg / mL, 32μg / mL, and 64μg / mL.

[0456] (measurement) 4.5 μL of the sample specimen was added to 180 μL of the first reagent incubated at 37° C., and allowed to react for 5 minutes at 37° C. Next, 45 μL of the second reagent was added to the mixture of the first reagent and the sample specimen, and allowed to react for an additional 5 minutes at 37° C. The absorbance Abs1 5 minutes after the sample specimen was added to the first reagent, and the absorbance Abs2 5 minutes after the second reagent was added were measured, and the absorbance change V was calculated according to the following formula (12). V=Abs2-Abs1×180 / 225 (12) Here, 180 represents the liquid volume of the first reagent, and 225 represents the total liquid volume of the first and second reagents.

[0457] The absorbance change measured using purified water instead of the sample specimen is used as the reagent blank, and the absorbance change V measured using the sample specimen is expressed as shown in the following formula (13). S Change in absorbance of the reagent blank from V B The value obtained by subtracting this was calculated as the sensitivity ΔAbs. ΔAbs=V S -V B (13)

[0458] As shown in the following formula (14), the sensitivity C measured using a sample specimen not containing calcium dobesilate was set as 100%, and the ratio of sensitivity B measured using a sample specimen containing calcium dobesilate was calculated as accuracy A. A = (B / C) × 100 (14) The results are shown in Table 5.

[0459] [Table 5]

[0460] As shown in Table 5, when a reagent without cyclodextrin or its analogues was used, the accuracy of the measurement was adversely affected by the addition of calcium dobesilate in a concentration-dependent manner. In contrast, by adding cyclodextrin or its analogues to the reagent, the effect of calcium dobesilate on the measurement was reduced, and the accuracy of the measurement was improved depending on the concentration of cyclodextrin or its analogues. Therefore, it was shown that in a measurement system using a redox color-developing reagent, the effect of calcium dobesilate contained in the sample on the measurement can be reduced by including α-cyclodextrin, β-cyclodextrin, or their analogues in the measurement reagent.

[0461] [Example 6: Effects of the type and concentration of cyclodextrin in a glycated albumin measurement reagent]

[0462] (First Reagent) The following reagent components were dissolved in purified water to the concentrations shown below, and the pH was adjusted to 7.6 to prepare a first reagent. In addition, a control reagent was prepared that did not contain cyclodextrin or its analogues. The cyclodextrin analogues used in Example 6 were methyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), hydroxypropyl-β-cyclodextrin (manufactured by Aldrich Co., Ltd.), trimethyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), and hydroxyethyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.). 100mmol / L N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid (Dojin Chemical Industries, Ltd.) 0.05% by weight / volume sodium azide (Merck) 1mmol / L Sodium ethylenediaminetetraacetate (Kokusan Chemical Co., Ltd.) 6% by weight / volume D(-)-sorbitol (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) 2mmol / L N,N-bis(4-sulfobutyl)-3-methylaniline disodium (Dojin Chemical Industries, Ltd.) 10U / mL peroxidase (Amano Enzyme Co., Ltd.) 30U / mL ketoamine oxidase (Asahi Kasei Pharma) 1%, 5%, 10%, 15%, or 20% w / v cyclodextrin or its analogues

[0463] (Second Reagent) Lucica GA-L GA-R2 (Asahi Kasei Pharma) containing protease and 4-AA was used.

[0464] (Sample specimen) Lucica GA-L Dedicated Control Serum L (Asahi Kasei Pharma) was prepared as the mother liquor for preparing the samples. Calcium dobesilate (Tokyo Kasei) was diluted with purified water to prepare a 640μg / mL aqueous solution of calcium dobesilate as the additive liquid for preparing the samples. The mother liquor was mixed with each additive liquid or purified water at a ratio of 9:1, and then mixed appropriately to obtain sample samples containing calcium dobesilate at concentrations of 0μg / mL, 16μg / mL, 32μg / mL, and 64μg / mL.

[0465] (measurement) 4.5 μL of the sample specimen was added to 180 μL of the first reagent incubated at 37° C., and allowed to react for 5 minutes at 37° C. Next, 45 μL of the second reagent was added to the mixture of the first reagent and the sample specimen, and allowed to react for an additional 5 minutes at 37° C. The absorbance Abs1 5 minutes after the sample specimen was added to the first reagent, and the absorbance Abs2 5 minutes after the second reagent was further added were measured, and the absorbance change V was calculated according to the following formula (15). V=Abs2-Abs1×180 / 225 (15) Here, 180 represents the liquid volume of the first reagent, and 225 represents the total liquid volume of the first and second reagents.

[0466] The absorbance change measured using purified water instead of the sample specimen is used as the reagent blank, and the absorbance change V measured using the sample specimen is calculated as shown in the following formula (16). S Change in absorbance of the reagent blank from V B The value obtained by subtracting this was calculated as the sensitivity ΔAbs. ΔAbs=V S -V B (16)

[0467] As shown in the following formula (17), the sensitivity C measured using a sample specimen not containing calcium dobesilate was set as 100%, and the ratio of sensitivity B measured using a sample specimen containing calcium dobesilate was calculated as accuracy A. A = (B / C) × 100 (17) The results are shown in Tables 6 and 7.

[0468] [Table 6] [Table 7]

[0469] As shown in Tables 6 and 7, when a reagent containing no cyclodextrin or its analogue was used, the accuracy of the measurement was adversely affected by the addition of calcium dobesilate in a concentration-dependent manner. In contrast, by adding a cyclodextrin analogue to the reagent, the effect of calcium dobesilate on the measurement was reduced, and the accuracy of the measurement was improved depending on the concentration of the cyclodextrin analogue. Therefore, it was shown that in a measurement system using a redox color-developing reagent, the effect of calcium dobesilate contained in the sample on the measurement can be reduced by including a cyclodextrin analogue in the measurement reagent.

[0470] [Example 7: Effect of cyclodextrin and sorbitol concentrations in glycated albumin measurement reagent]

[0471] (First Reagent) The following reagent components were dissolved in purified water to the concentrations shown below, and the pH was adjusted to 7.6 to prepare a first reagent. A control reagent was also prepared that did not contain cyclodextrin or its analogues. The cyclodextrin or its analogues used in Example 7 were methyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) and hydroxypropyl-β-cyclodextrin (manufactured by Aldrich Co., Ltd.). 100mmol / L N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid (Dojin Chemical Industries, Ltd.) 0.05% by weight / volume sodium azide (Merck) 1mmol / L Sodium ethylenediaminetetraacetate (Kokusan Chemical Co., Ltd.) 0 wt / vol% D(-)-sorbitol (Fujifilm Wako Pure Chemical Industries, Ltd.) or 1 wt / vol% D(-)-sorbitol (Fujifilm Wako Pure Chemical Industries, Ltd.) or 3 wt / vol% D(-)-sorbitol (Fujifilm Wako Pure Chemical Industries, Ltd.) or 6 wt / vol% D(-)-sorbitol (Fujifilm Wako Pure Chemical Industries, Ltd.) 2mmol / L N,N-bis(4-sulfobutyl)-3-methylaniline disodium (Dojin Chemical Industries, Ltd.) 10U / mL peroxidase (Amano Enzyme Co., Ltd.) 30U / mL ketoamine oxidase (Asahi Kasei Pharma) 10%, 15%, or 20% w / v cyclodextrin or its analogues

[0472] (Second Reagent) Lucica GA-L GA-R2 (Asahi Kasei Pharma) was used.

[0473] (Sample specimen) As the mother solution for preparing the samples, Lucica GA-L Special Control Serum L (Asahi Kasei Pharma) was prepared. As the additive solution for preparing the samples, calcium dobesilate (Tokyo Kasei) was diluted with purified water to prepare a 640μg / mL aqueous solution of calcium dobesilate. The mother solution was mixed with each additive solution or purified water at a ratio of 9:1, and then mixed appropriately to obtain sample samples containing calcium dobesilate at concentrations of 0μg / mL, 16μg / mL, 32μg / mL, and 64μg / mL.

[0474] (measurement) 4.5 μL of the sample specimen was added to 180 μL of the first reagent incubated at 37° C., and allowed to react for 5 minutes at 37° C. Next, 45 μL of the second reagent was added to the mixture of the first reagent and the sample specimen, and allowed to react for an additional 5 minutes at 37° C. The absorbance Abs1 5 minutes after the sample specimen was added to the first reagent, and the absorbance Abs2 5 minutes after the second reagent was added were measured, and the absorbance change V was calculated according to the following formula (18). V=Abs2-Abs1×180 / 225 (18) Here, 180 represents the liquid volume of the first reagent, and 225 represents the total liquid volume of the first and second reagents.

[0475] The absorbance change measured using purified water instead of the sample specimen is used as the reagent blank, and the absorbance change V measured using the sample specimen is expressed as shown in the following formula (19). S Change in absorbance of the reagent blank from V B The value obtained by subtracting this was calculated as the sensitivity ΔAbs. ΔAbs=V S -V B (19)

[0476] As shown in the following formula (20), the sensitivity C measured using a sample specimen not containing calcium dobesilate was set to 100%, and the percentage of sensitivity B measured using a sample specimen containing calcium dobesilate was calculated as accuracy A. A = (B / C) × 100 (20) The results are shown in Table 8.

[0477] [Table 8-1] [Table 8-2] [Table 8-3]

[0478] As shown in Table 8, when a reagent without cyclodextrin or its analogue was used, the accuracy of the measurement was adversely affected by the addition of calcium dobesilate in a concentration-dependent manner. In contrast, by adding cyclodextrin or its analogue to the reagent, the influence of calcium dobesilate on the measurement was reduced, and the accuracy of the measurement was improved depending on the concentration of cyclodextrin or its analogue. In addition, when a reagent without sorbitol was used, the accuracy of the measurement was adversely affected by the addition of calcium dobesilate in a concentration-dependent manner. In contrast, by adding sorbitol to the reagent, the influence of calcium dobesilate on the measurement was reduced, and the accuracy of the measurement was improved depending on the concentration of sorbitol. Therefore, it was shown that in a measurement system using a redox coloring reagent, the influence of calcium dobesilate contained in the sample on the measurement can be reduced by adding sorbitol to the measurement reagent composition.

[0479] [Example 8: Relationship between the effect of adding cyclodextrin or its analogue to a glycated albumin measurement reagent to avoid the influence of 2,5-dihydroxybenzenesulfonic acid and the reagent composition]

[0480] (First Reagent) The following reagent components were dissolved in purified water to the concentrations shown below and in Table 9, and the pH was adjusted to 7.6 to prepare a first reagent. A control reagent was also prepared that did not contain cyclodextrin or its analogue. The cyclodextrin or its analogue used in Example 8 was methyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.). 100mmol / L N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid (Dojin Chemical Industries, Ltd.) 1mmol / L Sodium ethylenediaminetetraacetate (Kokusan Chemical Co., Ltd.) 6% by weight / volume D(-)-sorbitol (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) 1.25mmol / L 4-aminoantipyrine (4-AA) (Actec) or 2mmol / L N,N-bis(4-sulfobutyl)-3-methylaniline disodium (TODB) (Dojin Chemical Industries, Ltd.) 0U / mL peroxidase or 10U / mL peroxidase (Amano Enzyme Co., Ltd.) 30U / mL ketoamine oxidase (Asahi Kasei Pharma) 5% by weight / volume of cyclodextrin or its analogues

[0481] (Second Reagent) The following reagent components were dissolved in purified water to the concentrations described below and in Table 9, and the pH was adjusted to 7.5 to prepare a second reagent. 200mmol / L 3,3-dimethylglutaric acid (Tokyo Chemical Industry Co., Ltd.) 0.05% by weight / volume Proclean 300 (manufactured by Sperco) 30% by weight / volume dimethyl sulfoxide (Nacalai Tesque) 0.1mmol / L Potassium ferrocyanide (Kokusan Chemical Co., Ltd.) 500U / mL catalase (Nagase Chemtex) 40 kU / mL protease for glycated albumin (manufactured by Asahi Kasei Pharma Corporation, obtained according to the method described in Patent No. 4565807) 5mmol / L 4-aminoantipyrine (4-AA) (Actec) or 8mmol / L N,N-bis(4-sulfobutyl)-3-methylaniline disodium (TODB) (Dojindo Chemical Industries, Ltd.) 0U / mL peroxidase or 40U / mL peroxidase (Amano Enzyme Co., Ltd.) As shown in Table 9, when the first reagent contained 4-AA, the second reagent contained TODB, and when the first reagent contained TODB, the second reagent contained 4-AA.

[0482] (Sample specimen) As the mother solution for preparing the samples, Lucica GA-L Special Control Serum L (Asahi Kasei Pharma) was prepared. As the additive solution for preparing the samples, calcium dobesilate (Tokyo Kasei) was diluted with purified water to prepare a 640μg / mL aqueous solution of calcium dobesilate. The mother solution was mixed with each additive solution or purified water at a ratio of 9:1, and then mixed appropriately to obtain sample samples containing calcium dobesilate at concentrations of 0μg / mL, 16μg / mL, 32μg / mL, and 64μg / mL.

[0483] (measurement) 4.5 μL of the sample specimen was added to 180 μL of the first reagent incubated at 37° C., and allowed to react for 5 minutes at 37° C. Next, 45 μL of the second reagent was added to the mixture of the first reagent and the sample specimen, and allowed to react for an additional 5 minutes at 37° C. The absorbance Abs1 5 minutes after the sample specimen was added to the first reagent, and the absorbance Abs2 5 minutes after the second reagent was added were measured, and the absorbance change V was calculated according to the following formula (21). V=Abs2-Abs1×180 / 225 (21) Here, 180 represents the liquid volume of the first reagent, and 225 represents the total liquid volume of the first and second reagents.

[0484] The absorbance change measured using purified water instead of the sample specimen is used as the reagent blank, and the absorbance change V measured using the sample specimen is calculated as shown in the following formula (22). S Change in absorbance of the reagent blank from V B The value obtained by subtracting this was calculated as the sensitivity ΔAbs. ΔAbs=V S -V B (twenty two)

[0485] As shown in the following formula (23), the sensitivity C measured using a sample specimen not containing calcium dobesilate was set to 100%, and the ratio of sensitivity B measured using a sample specimen containing calcium dobesilate was calculated as accuracy A. A = (B / C) × 100 (23) The results are shown in Table 10.

[0486] [Table 9] [Table 10]

[0487] As shown in Table 10, when a reagent without cyclodextrin or its analogues was used, the accuracy of the measurement was adversely affected by the addition of calcium dobesilate in a concentration-dependent manner. In contrast, by adding cyclodextrin or its analogues to the reagent, the effect of calcium dobesilate on the measurement was reduced, improving the accuracy of the measurement. In this case, the same improvement in accuracy by the addition of cyclodextrin or its analogues was confirmed in all four combinations of dye precursors and peroxidases.

[0488] [Example 9: Effect of adding cyclodextrin or its analogue to a creatinine measurement reagent to avoid the influence of 2,5-dihydroxybenzenesulfonic acid]

[0489] (First Reagent) Cyclodextrin was dissolved in Aqua Auto Kainos CRE-III plus reaction reagent I and Cygnus Auto CRE 7170 R1 to the concentration shown below to prepare the first reagent. The cyclodextrin or its analog used in Example 9 was hydroxypropyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.). 1%, 3%, or 5% w / v cyclodextrin or its analogues

[0490] (Second Reagent) Aqua Auto Kainos CRE-III plus reaction reagent II and Cygnus Auto CRE 7170 R2 were used as is.

[0491] (Sample specimen) A multi-item standard serum (Shinotestu) was prepared as the mother solution for preparing the samples. Calcium dobesilate (Tokyo Kasei) was diluted with purified water to prepare a 640 μg / mL aqueous solution of calcium dobesilate as the additive solution for preparing the samples. The mother solution was mixed with each additive solution or purified water at a ratio of 9:1, and then mixed appropriately to obtain sample samples containing calcium dobesilate at concentrations of 0 μg / mL, 16 μg / mL, 32 μg / mL, and 64 μg / mL.

[0492] (Measured using Aqua Auto Kainos CRE-III plus) 7.2 μL of the sample specimen was added to 180 μL of the first reagent incubated at 37° C., and allowed to react for 5 minutes at 37° C. Next, 60 μL of the second reagent was added to the mixture of the first reagent and the sample specimen, and allowed to react for an additional 5 minutes at 37° C. The absorbance Abs1 5 minutes after the sample specimen was added to the first reagent, and the absorbance Abs2 5 minutes after the second reagent was added were measured, and the absorbance change V was calculated according to the following formula (24). V=Abs2-Abs1×180 / 240 (24) Here, 180 represents the liquid volume of the first reagent, and 240 represents the total liquid volume of the first and second reagents.

[0493] (Measured using Cygnus Auto CRE 7170) 6.0 μL of the sample specimen was added to 150 μL of the first reagent incubated at 37° C., and allowed to react for 5 minutes at 37° C. Next, 50 μL of the second reagent was added to the mixture of the first reagent and the sample specimen, and allowed to react for an additional 5 minutes at 37° C. The absorbance Abs1 5 minutes after the sample specimen was added to the first reagent, and the absorbance Abs2 5 minutes after the second reagent was added were measured, and the absorbance change V was calculated according to the following formula (25). V=Abs2-Abs1×150 / 200 (25) Here, 150 represents the liquid volume of the first reagent, and 200 represents the total liquid volume of the first and second reagents.

[0494] The absorbance change measured using purified water instead of the sample specimen is used as the reagent blank, and the absorbance change V measured using the sample specimen is calculated as shown in the following formula (27). S Change in absorbance of the reagent blank from V B The value obtained by subtracting this was calculated as the sensitivity ΔAbs. ΔAbs=V S -V B (27)

[0495] As shown in the following formula (28), the sensitivity C measured using a sample specimen not containing calcium dobesilate was set to 100%, and the ratio of sensitivity B measured using a sample specimen containing calcium dobesilate was calculated as accuracy A. A = (B / C) × 100 (28) The results are shown in Table 11.

[0496] [Table 11-1] [Table 11-2]

[0497] As shown in Table 11, when a reagent without cyclodextrin or its analogues was used, the accuracy of the measurement was adversely affected by the addition of calcium dobesilate in a concentration-dependent manner. In contrast, the addition of cyclodextrin or its analogues to the reagent reduced the effect of calcium dobesilate on the measurement, improving the accuracy of the measurement. In this case, the improvement in accuracy by the addition of cyclodextrin or its analogues was confirmed in all of the creatinine measurement reagents used. Therefore, it was shown that in a measurement system using a redox coloring reagent, cyclodextrin and its analogues can reduce the effect of calcium dobesilate contained in the sample on the measurement.

[0498] [Example 10: Effect of adding cyclodextrin or its analogue to a uric acid measurement reagent to avoid the influence of 2,5-dihydroxybenzenesulfonic acid]

[0499] (First Reagent) Cyclodextrin was dissolved in Quick Auto UAII 7170 R1 (manufactured by Shino Test Co., Ltd.) to the concentration shown below to prepare the first reagent. The cyclodextrin or its analog used in Example 10 was hydroxypropyl-β-cyclodextrin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.). 5% by weight / volume of cyclodextrin or its analogues

[0500] (Second Reagent) A Quick Auto UAII 7170 R2 (manufactured by Sino-Test) was used as is.

[0501] (Sample specimen) A multi-item standard serum (Shinotes Co., Ltd.) was prepared as the mother solution for preparing the samples. Calcium dobesilate (Tokyo Kasei Co., Ltd.) was diluted with purified water to prepare a 640 μg / mL aqueous solution of calcium dobesilate as the additive solution for preparing the samples. The mother solution was mixed with each additive solution or purified water at a ratio of 9:1, and then mixed appropriately to obtain sample specimens containing calcium dobesilate at concentrations of 0 μg / mL and 64 μg / mL.

[0502] (Measured using Quick Auto UAII 7170) 3.3 μL of the sample specimen was added to 160 μL of the first reagent incubated at 37° C., and allowed to react for 5 minutes at 37° C. Next, 40 μL of the second reagent was added to the mixture of the first reagent and the sample specimen, and allowed to react for an additional 5 minutes at 37° C. The absorbance Abs1 5 minutes after the sample specimen was added to the first reagent, and the absorbance Abs2 5 minutes after the second reagent was further added were measured, and the absorbance change V was calculated according to the following formula (29). V=Abs2-Abs1×160 / 200 (29) Here, 160 represents the liquid volume of the first reagent, and 200 represents the total liquid volume of the first and second reagents.

[0503] The absorbance change measured using purified water instead of the sample specimen is used as the reagent blank, and the absorbance change V measured using the sample specimen is calculated as shown in the following formula (30). S Change in absorbance of the reagent blank from V B The value obtained by subtracting this was calculated as the sensitivity ΔAbs. ΔAbs=V S -V B (30)

[0504] As shown in the following formula (31), the sensitivity C measured using a sample specimen not containing calcium dobesilate was set to 100%, and the ratio of sensitivity B measured using a sample specimen containing calcium dobesilate was calculated as accuracy A. A = (B / C) × 100 (31) The results are shown in Table 12.

[0505] [Table 12]

[0506] As shown in Table 12, when a reagent without cyclodextrin or its analogue was used, the accuracy of the measurement was adversely affected by the addition of calcium dobesilate. In contrast, by adding cyclodextrin or its analogue to the reagent, the effect of calcium dobesilate on the measurement was reduced, and the accuracy of the measurement was improved. In addition to Example 9, the improvement in accuracy by adding cyclodextrin or its analogue was also confirmed in the uric acid measurement reagent. Therefore, it was shown that in a measurement system using an oxidation-reduction coloring reagent, cyclodextrin and its analogue can reduce the effect of calcium dobesilate contained in the sample on the measurement.

Claims

1. Peroxidase and A color-developing reagent configured to react with hydrogen peroxide derived from a substance in a sample in the presence of the peroxidase to develop a color; Cyclodextrin or an analog thereof; A reagent composition for measurement comprising: The cyclodextrin or its analogue suppresses color development inhibition caused by 2,5-dihydroxybenzenesulfonic acid or a salt thereof which may be contained in the sample. Measurement reagent composition.

2. The reagent composition according to claim 1 , wherein the color-developing reagent comprises a Trinder reagent and a coupler.

3. The reagent composition for measurement according to claim 2, wherein the reagent composition for measurement comprises a first reagent and a second reagent, the first reagent comprises the coupler and the cyclodextrin or an analog thereof, and the second reagent comprises the Trinder reagent and the peroxidase.

4. The reagent composition for measurement according to claim 2, wherein the reagent composition for measurement comprises a first reagent and a second reagent, the first reagent comprises the coupler, the peroxidase, and the cyclodextrin or an analog thereof, and the second reagent comprises the Trinder reagent.

5. The reagent composition for measurement according to claim 2, wherein the reagent composition for measurement comprises a first reagent and a second reagent, the first reagent comprises the Trinder reagent and the cyclodextrin or an analog thereof, and the second reagent comprises the coupler and the peroxidase.

6. The reagent composition for measurement according to claim 2, wherein the reagent composition for measurement comprises a first reagent and a second reagent, the first reagent comprises the Trinder reagent, the peroxidase, and the cyclodextrin or an analog thereof, and the second reagent comprises the coupler.

7. The reagent composition for measurement according to claim 3 , wherein the first reagent is added to the sample, and then the second reagent is added to the sample.

8. The reagent composition for measurement according to claim 3 , wherein the first reagent and the second reagent are packaged separately.

9. The reagent composition according to claim 1 , wherein the color-developing reagent is a leuco dye.

10. 10. The reagent composition according to claim 1, wherein the cyclodextrin is at least one selected from the group consisting of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin.

11. The reagent composition according to claim 1 , wherein the concentration of the cyclodextrin or its analog in the reagent composition is 0.1% by weight / volume to 35% by weight / volume.

12. The reagent composition according to claim 1 , wherein the substance in the sample is glycated albumin, uric acid, or creatinine.

13. The reagent composition according to claim 1 , further comprising ferrocyanide.

14. The reagent composition according to claim 1 , further comprising a sugar alcohol other than the cyclodextrin or its analogue.

15. The reagent composition according to any one of claims 1 to 14, wherein the 2,5-dihydroxybenzenesulfonic acid or a salt thereof is at least one selected from the group consisting of calcium dobesilate and ethamsylate.

16. A measurement reagent composition is provided, the measurement reagent composition comprising: peroxidase; a color-developing reagent configured to react with hydrogen peroxide derived from a substance in a sample in the presence of the peroxidase to develop a color; and cyclodextrin or an analog thereof; mixing the reagent composition for measurement with the sample and measuring color development; A method for measuring a substance in a sample, comprising: The sample contains 2,5-dihydroxybenzenesulfonic acid or a salt thereof. A method for measuring a substance in a sample.

17. The method for measuring a substance in a sample according to claim 16, wherein the color-developing reagent comprises a Trinder reagent and a coupler.

18. The method for measuring a substance in a sample according to claim 17, wherein the measurement reagent composition comprises a first reagent and a second reagent, the first reagent comprises the coupler and the cyclodextrin or an analog thereof, and the second reagent comprises the Trinder reagent and the peroxidase.

19. The method for measuring a substance in a sample according to claim 17, wherein the measurement reagent composition comprises a first reagent and a second reagent, the first reagent comprises the coupler, the peroxidase, and the cyclodextrin or an analog thereof, and the second reagent comprises the Trinder reagent.

20. The method for measuring a substance in a sample according to claim 17, wherein the measurement reagent composition comprises a first reagent and a second reagent, the first reagent comprises the Trinder reagent and the cyclodextrin or an analog thereof, and the second reagent comprises the coupler and the peroxidase.

21. The method for measuring a substance in a sample according to claim 17, wherein the measurement reagent composition comprises a first reagent and a second reagent, the first reagent comprises the Trinder reagent, the peroxidase, and the cyclodextrin or an analog thereof, and the second reagent comprises the coupler.

22. The method for measuring a substance in a sample according to claim 18 , further comprising mixing the first reagent with the sample and then mixing the second reagent with the sample.

23. 23. The method for measuring a substance in a sample according to claim 18, wherein the first reagent and the second reagent are packaged separately in the reagent composition for measurement.

24. The method for measuring a substance in a sample according to claim 16, wherein the color-developing reagent is a leuco dye.

25. 25. The method for measuring a substance in a sample according to any one of claims 16 to 24, wherein the cyclodextrin is at least one selected from the group consisting of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin.

26. 26. The method for measuring a substance in a sample according to any one of claims 16 to 25, wherein the concentration of the cyclodextrin or its analogue when the measurement reagent composition and the sample are mixed is 0.1 weight / volume % to 35 weight / volume %.

27. The method for measuring a substance in a sample according to any one of claims 16 to 26, wherein the substance in the sample is glycated albumin, uric acid, or creatinine.

28. The method for measuring a substance in a sample according to any one of claims 16 to 27, wherein the reagent composition for measurement further comprises ferrocyanide.

29. 29. The method for measuring a substance in a sample according to claim 16, wherein the reagent composition for measurement further comprises a sugar alcohol other than the cyclodextrin or an analog thereof.

30. The method for measuring a substance in a sample according to any one of claims 16 to 29, wherein the 2,5-dihydroxybenzenesulfonic acid or a salt thereof is at least one selected from the group consisting of calcium dobesilate and ethamsylate.

31. An inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof, comprising a cyclodextrin or an analog thereof, Inhibits the inhibition of a color-developing reaction using a color-developing reagent by 2,5-dihydroxybenzenesulfonic acid or a salt thereof, The color-developing reagent is a color-developing reagent that reacts with hydrogen peroxide in the presence of peroxidase to develop color. An inhibitor of 2,5-dihydroxybenzenesulfonic acid or its salt.

32. The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to claim 31, wherein the color-developing reagent is at least one selected from the group consisting of a Trinder reagent and a coupler.

33. The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to claim 31, wherein the color-developing reagent is a leuco dye.

34. The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to any one of claims 31 to 33, which inhibits the action of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on a substance.

35. The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to any one of claims 31 to 34, which inhibits 2,5-dihydroxybenzenesulfonic acid or a salt thereof from inhibiting a color-developing reaction using the color-developing reagent.

36. The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to any one of claims 31 to 35, wherein the 2,5-dihydroxybenzenesulfonic acid or a salt thereof is at least one selected from the group consisting of calcium dobesilate and ethamsylate.

37. The inhibitor of 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to any one of claims 31 to 36, wherein the cyclodextrin is at least one selected from the group consisting of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin.

38. The method includes mixing cyclodextrin or an analog thereof with 2,5-dihydroxybenzenesulfonic acid or a salt thereof.

1. A method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof, comprising: By mixing, the inhibition of the color-developing reaction using a color-developing reagent by 2,5-dihydroxybenzenesulfonic acid or a salt thereof is prevented, The color-developing reagent is a color-developing reagent that reacts with hydrogen peroxide in the presence of peroxidase to develop color. method.

39. The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to claim 38, wherein the color-developing reagent is at least one selected from the group consisting of Trinder's reagent and a coupler.

40. The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to claim 38, wherein the color-developing reagent is a leuco dye.

41. The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to any one of claims 38 to 40, wherein the mixing inhibits the action of 2,5-dihydroxybenzenesulfonic acid or a salt thereof on a substance.

42. The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to any one of claims 38 to 41, wherein the mixing inhibits 2,5-dihydroxybenzenesulfonic acid or a salt thereof from inhibiting a color-developing reaction using the color-developing reagent.

43. The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to any one of claims 38 to 42, wherein the 2,5-dihydroxybenzenesulfonic acid or a salt thereof is at least one selected from the group consisting of calcium dobesilate and ethamsylate.

44. The method for inhibiting 2,5-dihydroxybenzenesulfonic acid or a salt thereof according to any one of claims 38 to 43, wherein the cyclodextrin is at least one selected from the group consisting of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin.

45. The method includes using cyclodextrin or an analogue thereof to suppress inhibition of color development caused by 2,5-dihydroxybenzenesulfonic acid or a salt thereof in a reaction between hydrogen peroxide derived from a substance in a sample and a color-developing reagent in the presence of peroxidase. A method for suppressing color development inhibition caused by 2,5-dihydroxybenzenesulfonic acid or a salt thereof.

Images