WO2012020744A1 - 糖化ヘモグロビンの測定方法 - Google Patents
糖化ヘモグロビンの測定方法 Download PDFInfo
- Publication number
- WO2012020744A1 WO2012020744A1 PCT/JP2011/068103 JP2011068103W WO2012020744A1 WO 2012020744 A1 WO2012020744 A1 WO 2012020744A1 JP 2011068103 W JP2011068103 W JP 2011068103W WO 2012020744 A1 WO2012020744 A1 WO 2012020744A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- reagent
- hemoglobin
- measurement
- kit
- examples
- Prior art date
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/72—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood pigments, e.g. haemoglobin, bilirubin or other porphyrins; involving occult blood
- G01N33/721—Haemoglobin
- G01N33/723—Glycosylated haemoglobin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
- C12Q1/28—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase involving peroxidase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
- C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
Definitions
- the present invention relates to a method for measuring glycated hemoglobin, a measuring reagent, and a measuring kit.
- Glycated hemoglobin is a saccharified product in which glucose is bound to hemoglobin.
- Hemoglobin has a tetramer structure consisting of ⁇ and ⁇ chains, but the glycated N-terminus of ⁇ chain is called hemoglobin A1c, and increases as the blood glucose level increases. Measured in clinical tests.
- an immunoassay method using an antibody such as a chromatography method such as HPLC, electrophoresis, latex immunoagglutination, an enzyme that acts on glycated protein, and a glycated peptide and / or glycated amino acid.
- An enzyme measurement method using an enzyme is known.
- glycated hemoglobin As an enzyme measurement method for glycated hemoglobin, first, hemoglobin in a hemoglobin-containing sample is denatured with a denaturing agent, a proteolytic enzyme is allowed to act on the denatured hemoglobin, and then a glycated peptide oxidase is allowed to act on the produced glycated peptide. It is known to react the oxidized hydrogen peroxide with an oxidative coloring type chromogen in the presence of a peroxidase active substance such as peroxidase, lead to the dye, and measure the glycated hemoglobin from the absorbance of the dye produced (absorbance method). It has been.
- Patent Document 1 a method using a cationic surfactant and / or an amphoteric surfactant
- Patent Document 2 a method using a sulfone compound and / or a nitro compound
- Patent Document 4 a method using a tetrazolium compound
- Patent Document 5 a method using a specific anionic surfactant such as polyoxyethylene alkyl ether sulfates
- An object of the present invention is to provide a method and a reagent for accurately and highly sensitively measuring glycated hemoglobin in a hemoglobin-containing sample without being affected by hemoglobin.
- the present inventors conducted a reaction in which a proteolytic enzyme is allowed to act on a hemoglobin-containing sample in the presence of a surfactant and then a fructosyl peptide oxidase is allowed to act in the presence of an isothiazolinone derivative.
- the inventors have found the knowledge that by measuring the generated hydrogen peroxide, glycated hemoglobin in a hemoglobin-containing sample can be measured accurately and with high sensitivity without being affected by hemoglobin, and the present invention has been completed. That is, the present invention relates to the following [1] to [15].
- a 1 represents a hydrogen atom or substituted or unsubstituted alkyl
- a 2 and A 3 are the same or different and each represents a hydrogen atom, substituted or unsubstituted alkyl, halogen atom, or Together form a ring structure
- the compound represented by formula (I) is 2-alkyl-4-isothiazolin-3-one, 1,2-benzisothiazol-3 (2H) -one, and 2-alkyl-4,5-
- the measuring method according to [2] which is an isothiazolinone derivative selected from the group consisting of dihalogeno-4-isothiazolin-3-one.
- a reagent for measuring glycated hemoglobin in a hemoglobin-containing sample comprising a proteolytic enzyme, a fructosyl peptide oxidase, an isothiazolinone derivative, and a surfactant.
- a 1 represents a hydrogen atom or substituted or unsubstituted alkyl
- a 2 and A 3 are the same or different and each represents a hydrogen atom, substituted or unsubstituted alkyl, halogen atom, or Together form a ring structure
- the compound represented by the formula (I) is 2-alkyl-4-isothiazolin-3-one, 1,2-benzisothiazol-3 (2H) -one, and 2-alkyl-4,5-
- the measuring reagent according to [7] which is an isothiazolinone derivative selected from the group consisting of dihalogeno-4-isothiazolin-3-one.
- a kit for measuring glycated hemoglobin in a hemoglobin-containing sample comprising: a first reagent containing a proteolytic enzyme, an isothiazolinone derivative, and a surfactant; and a second reagent containing a fructosyl peptide oxidase
- a kit for measuring glycated hemoglobin comprising: [12] A kit for measuring glycated hemoglobin in a hemoglobin-containing sample, a first reagent containing a proteolytic enzyme and a surfactant, a fructosyl peptide oxidase, and a second reagent containing an isothiazolinone derivative
- the measurement kit according to [11] or [12], wherein the isothiazolinone derivative is a compound represented by the following formula (I).
- a 1 represents a hydrogen atom or substituted or unsubstituted alkyl
- a 2 and A 3 are the same or different and each represents a hydrogen atom, a substituted or unsubstituted alkyl, or a halogen atom, Or together form a ring structure
- the compound represented by formula (I) is 2-alkyl-4-isothiazolin-3-one, 1,2-benzisothiazol-3 (2H) -one, and 2-alkyl-4,5-
- the measurement kit according to [13] which is an isothiazolinone derivative selected from the group consisting of dihalogeno-4-isothiazolin-3-one.
- a method, a reagent, and a kit for accurately and highly sensitively measuring glycated hemoglobin in a hemoglobin-containing sample without being affected by hemoglobin are provided.
- ⁇ represents the kit of Comparative Example 5
- ⁇ represents the kit of Example 9
- ⁇ represents the measurement using the kit of Example 10
- the vertical axis represents the reaction absorbance ( ⁇ 10 ⁇ 4 Abs)
- the horizontal axis Represents the hemoglobin concentration (mg / mL).
- ⁇ represents the kit of Comparative Example 4
- ⁇ represents the kit of Example 7
- ⁇ represents the measurement using the kit of Example 8
- the vertical axis represents the reaction absorbance ( ⁇ 10 ⁇ 4 Abs)
- the horizontal axis Represents the hemoglobin concentration (mg / mL).
- ⁇ represents the kit of Comparative Example 11
- ⁇ represents the kit of Example 21
- ⁇ represents the measurement using the kit of Example 22
- the vertical axis represents the reaction absorbance ( ⁇ 10 ⁇ 4 Abs)
- the horizontal axis Represents the hemoglobin concentration (mg / mL).
- a proteolytic enzyme acts on glycated hemoglobin in a hemoglobin-containing sample in the presence of a surfactant. Then, in a reaction in which fructosyl peptide oxidase is allowed to act, the reaction is performed in the presence of an isothiazolinone derivative, and the hydrogen peroxide produced is measured.
- the isothiazolinone derivative may be present in a reaction in which fructosyl peptide oxidase acts, or may be present in both a reaction in which proteolytic enzyme acts and a reaction in which fructosyl peptide oxidase acts. Specifically, a measurement method including the following steps is exemplified.
- ⁇ Measurement method 1> (1) A step of allowing a proteolytic enzyme to act on glycated hemoglobin in a hemoglobin-containing sample in the presence of a surfactant; (2) A step of causing a fructosyl peptide oxidase to act on the reaction product obtained in step (1) in the presence of an isothiazolinone derivative to generate hydrogen peroxide; (3) measuring hydrogen peroxide produced in step (2); and (4) Based on a calibration curve representing the relationship between the amount of hydrogen peroxide and the concentration of glycated hemoglobin prepared in advance using a known concentration of glycated hemoglobin, from the amount of hydrogen peroxide measured in step (3), the hemoglobin-containing sample The step of determining the glycated hemoglobin concentration.
- ⁇ Measurement method 2> A step of allowing a proteolytic enzyme to act on glycated hemoglobin in a hemoglobin-containing sample in the presence of an isothiazolinone derivative and a surfactant; (2) reacting the reaction product obtained in step (1) with fructosyl peptide oxidase to generate hydrogen peroxide; (3) measuring hydrogen peroxide produced in step (2); and (4) Based on a calibration curve representing the relationship between the amount of hydrogen peroxide and the concentration of glycated hemoglobin prepared in advance using a known concentration of glycated hemoglobin, from the amount of hydrogen peroxide measured in step (3), the hemoglobin-containing sample The step of determining the glycated hemoglobin concentration.
- the method for measuring glycated hemoglobin in the hemoglobin-containing sample of the present invention includes a method for calculating the ratio of the amount of glycated hemoglobin in the hemoglobin-containing sample to the total hemoglobin (that is, the total hemoglobin in which hemoglobin and glycated hemoglobin are combined). Include.
- the method for measuring glycated hemoglobin in the hemoglobin-containing sample of the present invention is specifically a measurement method including the following steps.
- ⁇ Measurement method 3> (1) determining the amount of total hemoglobin in the hemoglobin-containing sample (that is, total hemoglobin obtained by combining hemoglobin and glycated hemoglobin); (2) a step of allowing a proteolytic enzyme to act on glycated hemoglobin in a hemoglobin-containing sample in the presence of a surfactant; (3) a step of causing the reaction product obtained in step (2) to act on a fructosyl peptide oxidase in the presence of an isothiazolinone derivative to generate hydrogen peroxide; (4) measuring hydrogen peroxide produced in step (3); and (5) Based on a calibration curve representing the relationship between the amount of hydrogen peroxide and the amount of glycated hemoglobin prepared in advance using a known amount of glycated hemoglobin, from the amount of hydrogen peroxide measured in step (4), the hemoglobin-containing sample Determining the amount of glycated hemoglobin of; and (6) A step of
- ⁇ Measurement method 4> (1) determining the amount of total hemoglobin in the hemoglobin-containing sample (that is, total hemoglobin obtained by combining hemoglobin and glycated hemoglobin); (2) a step of allowing a proteolytic enzyme to act on glycated hemoglobin in a hemoglobin-containing sample in the presence of an isothiazolinone derivative and a surfactant; (3) a step of causing a fructosyl peptide oxidase to act on the reaction product obtained in step (2) to generate hydrogen peroxide; (4) measuring hydrogen peroxide produced in step (3); and (5) Based on a calibration curve representing the relationship between the amount of hydrogen peroxide and the amount of glycated hemoglobin prepared in advance using a known amount of glycated hemoglobin, from the amount of hydrogen peroxide measured in step (4), the hemoglobin-containing sample Determining the amount of glycated hemoglobin of; and (6) A step of calculating a
- the hemoglobin-containing sample in the measurement method of the present invention is not particularly limited as long as it contains hemoglobin and can be applied to the measurement method of glycated hemoglobin of the present invention.
- whole blood, blood cells, and blood cells mixed with plasma And samples obtained by hemolyzing these samples.
- the hemolysis treatment is not particularly limited as long as it is a treatment that hemolyzes whole blood, blood cells, and a sample in which blood cells are mixed with plasma, and examples thereof include physical methods, chemical methods, and biological methods.
- Examples of the physical method include a method using a hypotonic solution such as distilled water and a method using ultrasonic waves.
- Examples of the chemical method include a method using an organic solvent such as methanol, ethanol, and acetone, and a method using a polyoxyethylene surfactant.
- biological methods include a method using an antibody or complement.
- the glycated hemoglobin in the present invention is produced by binding sugar such as glucose to hemoglobin, and examples thereof include hemoglobin A1a, hemoglobin A1b, hemoglobin A1c, and hemoglobin A1c is preferable.
- the isothiazolinone derivative in the present invention is not particularly limited as long as it is an isothiazolinone derivative that enables the method for measuring glycated hemoglobin of the present invention.
- a compound represented by the following formula (I) [hereinafter referred to as compound (I) ].
- a 1 represents a hydrogen atom or substituted or unsubstituted alkyl
- a 2 and A 3 are the same or different and each represents a hydrogen atom, a substituted or unsubstituted alkyl, or a halogen atom, Or together form a ring structure
- a 1 in the compound (I) represents a substituted or unsubstituted alkyl
- a 2 and A 3 are the same or different and each represents a hydrogen atom, a substituted or unsubstituted alkyl, a halogen atom, or together.
- Examples of the alkyl in the substituted or unsubstituted alkyl include linear alkyl having 1 to 20 carbon atoms and branched alkyl having 3 to 20 carbon atoms.
- linear alkyl having 1 to 20 carbon atoms examples include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl (lauryl), tridecyl, tetradecyl (myristyl), pentadecyl, hexadecyl (Cetyl), heptadecyl, octadecyl (stearyl), nonadecyl, icosyl and the like.
- Examples of the branched alkyl having 3 to 20 carbon atoms include isopropyl, isobutyl, isopentyl, isohexyl, isoheptyl, isooctyl, isononyl, isodecyl, isoundecyl, isododecyl, isotridecyl, isotetradecyl, isopentadecyl, isohexadecyl, isoheptadecyl, isooctadecyl , Isononadecyl, isoicosyl, octyldodecyl and the like.
- halogen atom examples include a chlorine atom, a bromine atom, and an iodine atom.
- substituent in the substituted alkyl include a phenyl group, a hydroxyl group, a sulfo group, a cyano group, and a halogen atom.
- a halogen atom the above-mentioned halogen atom etc. are mentioned, for example.
- Examples of the ring structure formed by combining A 2 and A 3 include a benzene ring and a naphthalene ring.
- compound (I) examples include 2-alkyl-4-isothiazolin-3-one, 1,2-benzisothiazol-3 (2H) -one, 2-alkyl-4,5-dihalogeno-4- And isothiazoline-3-one.
- Examples of commercially available compounds (I) include 2-octyl-4-isothiazolin-3-one (manufactured by Tokyo Chemical Industry Co., Ltd.), 1,2-benzisothiazol-3 (2H) -one (manufactured by Wako Pure Chemical Industries, Ltd.) 4,5-dichloro-2-octyl-4-isothiazolin-3-one (manufactured by Hichem) and the like.
- the concentration of the isothiazolinone derivative in the reaction solution is not particularly limited as long as it allows the method for measuring glycated hemoglobin according to the present invention, but is usually 0.005 to 20 mmol. / L, preferably 0.01 to 10 mmol / L.
- the surfactant in the present invention is not particularly limited as long as it is a surfactant that enables the method for measuring glycated hemoglobin of the present invention.
- a cationic surfactant for example, a cationic surfactant, an anionic surfactant, an amphoteric surfactant Agents, nonionic surfactants and the like.
- cationic surfactant examples include quaternary ammonium salts, pyridinium salts, phosphonium salts, imidazolium salts, isoquinolinium salts, and the like, and quaternary ammonium salts, pyridinium salts, and phosphonium salts are preferable.
- quaternary ammonium salt having at least one linear alkyl having 8 to 20 carbon atoms is preferable.
- linear alkyl having 8 to 20 carbon atoms include octyl, nonyl, decyl, undecyl, dodecyl (lauryl), tridecyl, tetradecyl (myristyl), pentadecyl, hexadecyl (cetyl), heptadecyl, octadecyl (stearyl), nonadecyl, icosyl Etc.
- Specific examples (products) of the quaternary ammonium salt include, for example, decyltrimethylammonium chloride, decyltrimethylammonium bromide (hereinafter referred to as C10TMA), dodecyltrimethylammonium chloride, dodecyltrimethylammonium bromide, hexadecyltrimethylammonium chloride, hexadecyl.
- C10TMA decyltrimethylammonium chloride
- C10TMA decyltrimethylammonium bromide
- dodecyltrimethylammonium chloride dodecyltrimethylammonium bromide
- hexadecyltrimethylammonium chloride dodecyltrimethylammonium bromide
- hexadecyltrimethylammonium chloride hexadecyl.
- Examples thereof include trimethylammonium bromide, didecyldimethylammonium chloride, didecyldimethylammonium bromide, didodecyldimethylammonium chloride, didodecyldimethylammonium bromide (all manufactured by Tokyo Chemical Industry Co., Ltd.).
- pyridinium salt a pyridinium salt represented by the following general formula (II) [hereinafter referred to as compound (II)] is used.
- R 1 is substituted or unsubstituted alkyl, or substituted or unsubstituted alkenyl
- R a is a hydrogen atom, substituted or unsubstituted alkyl, or substituted or unsubstituted alkenyl
- n is 1 to 5
- X ⁇ represents a monovalent anion.
- Examples of the alkyl in the substituted or unsubstituted alkyl in R 1 include linear alkyl having 1 to 20 carbon atoms, branched alkyl having 3 to 20 carbon atoms, and the like. A branched alkyl having 8 to 20 is preferable.
- linear alkyl having 1 to 20 carbon atoms examples include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl (lauryl), tridecyl, tetradecyl (myristyl), pentadecyl, hexadecyl (Cetyl), heptadecyl, octadecyl (stearyl), nonadecyl, icosyl and the like.
- Examples of the branched alkyl having 3 to 20 carbon atoms include isopropyl, isobutyl, isopentyl, isohexyl, isoheptyl, isooctyl, isononyl, isodecyl, isoundecyl, isododecyl, isotridecyl, isotetradecyl, isopentadecyl, isohexadecyl, isoheptadecyl, isooctadecyl , Isononadecyl, isoicosyl, octyldodecyl and the like.
- Examples of the linear alkyl having 8 to 20 carbon atoms include the aforementioned linear alkyl having 8 to 20 carbon atoms.
- Examples of the branched alkyl having 8 to 20 carbon atoms include isooctyl, isononyl, isodecyl, isoundecyl, isododecyl, isotridecyl, isotetradecyl, isopentadecyl, isohexadecyl, isoheptadecyl, isooctadecyl, isononadecyl, isoicosyl, octyldodecyl and the like. It is done.
- the alkenyl in the substituted or unsubstituted alkenyl includes, for example, alkenyl having 2 to 20 carbon atoms, and preferably alkenyl having 8 to 20 carbon atoms.
- alkenyl having 2 to 20 carbon atoms include vinyl, propenyl, allyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, oleyl, nonadecenyl, Etc.
- alkenyl having 8 to 20 carbon atoms examples include octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, oleyl, nonadecenyl, icocenyl and the like.
- examples of the substituent in the substituted alkyl and the substituted alkenyl include a phenyl group, a hydroxyl group, a sulfo group, a cyano group, and a halogen atom.
- examples of the halogen atom include a chlorine atom, a bromine atom, and an iodine atom.
- examples of the alkyl in the substituted or unsubstituted alkyl include linear alkyl having 1 to 20 carbon atoms and branched alkyl having 3 to 20 carbon atoms.
- examples of the linear alkyl having 1 to 20 carbon atoms include the aforementioned linear alkyl having 1 to 20 carbon atoms.
- examples of the branched alkyl having 3 to 20 carbon atoms include the aforementioned branched alkyl having 3 to 20 carbon atoms.
- the alkenyl in the substituted or unsubstituted alkenyl includes, for example, alkenyl having 2 to 20 carbon atoms.
- alkenyl having 2 to 20 carbon atoms include the above-mentioned linear alkenyl having 2 to 20 carbon atoms.
- examples of the substituent in the substituted alkyl and the substituted alkenyl include a phenyl group, a hydroxyl group, a sulfo group, a cyano group, and a halogen atom.
- examples of the phenyl group-substituted alkyl include benzyl and 1-phenylethyl.
- examples of the halogen atom include a chlorine atom, a bromine atom, and an iodine atom.
- X in the compound (II) - represents a monovalent anion.
- monovalent anions include halogen ions, OH ⁇ , PF 6 ⁇ , BF 4 ⁇ , CH 3 CH 2 OSO 3 ⁇ , (CF 3 SO 2 ) 2 N ⁇ and the like.
- halogen ion include Cl ⁇ , Br ⁇ , I ⁇ and the like.
- compound (II) examples include, for example, 1-dodecylpyridinium chloride (hereinafter referred to as C12py; manufactured by Tokyo Chemical Industry Co., Ltd.), 1-cetylpyridinium chloride, 1-cetyl-4-methylpyridinium chloride (Tokyo Chemical Industry). And N-octadecyl-4-stilbazole bromide (manufactured by Tokyo Chemical Industry Co., Ltd.).
- C12py 1-dodecylpyridinium chloride
- C12py 1-cetylpyridinium chloride
- 1-cetyl-4-methylpyridinium chloride Tokyo Chemical Industry
- N-octadecyl-4-stilbazole bromide manufactured by Tokyo Chemical Industry Co., Ltd.
- a phosphonium salt represented by the following general formula (III) [hereinafter referred to as compound (III)] is used.
- R 2 to R 5 are the same or different and each represents a substituted or unsubstituted alkyl, and Y ⁇ represents a monovalent anion.
- examples of the alkyl in the substituted or unsubstituted alkyl include linear alkyl having 8 to 20 carbon atoms and branched alkyl having 8 to 20 carbon atoms.
- Examples of the linear alkyl having 8 to 20 carbon atoms include the aforementioned linear alkyl having 8 to 20 carbon atoms.
- Examples of the branched alkyl having 8 to 20 carbon atoms include the aforementioned branched alkyl having 8 to 20 carbon atoms.
- the substituent in the substituted alkyl include a phenyl group, a hydroxyl group, a sulfo group, a cyano group, and a halogen atom.
- Examples of the phenyl group-substituted alkyl include benzyl and 1-phenylethyl.
- Examples of the halogen atom include a chlorine atom, a bromine atom, and an iodine atom.
- examples of the alkyl in the substituted or unsubstituted alkyl include linear alkyl having 1 to 20 carbon atoms and branched alkyl having 3 to 20 carbon atoms.
- Examples of the linear alkyl having 1 to 20 carbon atoms include the aforementioned linear alkyl having 1 to 20 carbon atoms.
- Examples of the branched alkyl having 3 to 20 carbon atoms include the aforementioned branched alkyl having 3 to 20 carbon atoms.
- the substituent in the substituted alkyl include a phenyl group, a hydroxyl group, a sulfo group, a cyano group, and a halogen atom.
- Examples of the phenyl group-substituted alkyl include benzyl and 1-phenylethyl.
- Examples of the halogen atom include a chlorine atom, a bromine atom, and an iodine atom.
- Y ⁇ represents a monovalent anion.
- monovalent anions include halogen ions, OH ⁇ , PF 6 ⁇ , BF 4 ⁇ , CH 3 CH 2 OSO 3 ⁇ , (CF 3 SO 2 ) 2 N ⁇ , B (C 6 H 5 ) 4 ⁇ , And anions such as benzotriazolate.
- halogen ion include Cl ⁇ , Br ⁇ , I ⁇ and the like.
- compound (III) examples include tetraoctylphosphonium bromide (manufactured by Tokyo Chemical Industry Co., Ltd.), tributyloctylphosphonium bromide (manufactured by Tokyo Chemical Industry Co., Ltd.), tributyldodecylphosphonium bromide, tributylhexadecylphosphonium bromide and the like. .
- an imidazolium salt [hereinafter referred to as compound (IV)] represented by the following general formula (IV) is used.
- R 6 and R 8 are the same or different and each represents substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl
- R 7 , R 9 , and R 10 are hydrogen atoms, substituted or unsubstituted alkyl, substituted or represents unsubstituted alkenyl
- Z - represents a monovalent anion
- examples of the alkyl in the substituted or unsubstituted alkyl include linear alkyl having 8 to 20 carbon atoms and branched alkyl having 8 to 20 carbon atoms.
- examples of the linear alkyl having 8 to 20 carbon atoms include the aforementioned linear alkyl having 8 to 20 carbon atoms.
- examples of the branched alkyl having 8 to 20 carbon atoms include the aforementioned branched alkyl having 8 to 20 carbon atoms.
- the alkenyl in the substituted or unsubstituted alkenyl includes, for example, alkenyl having 8 to 20 carbon atoms.
- alkenyl having 8 to 20 carbon atoms include the aforementioned alkenyl having 8 to 20 carbon atoms.
- examples of the substituent in the substituted alkyl and the substituted alkenyl include a phenyl group, a hydroxyl group, a sulfo group, a cyano group, and a halogen atom.
- examples of the phenyl group-substituted alkyl include benzyl and 1-phenylethyl.
- examples of the halogen atom include a chlorine atom, a bromine atom, and an iodine atom.
- examples of the alkyl in the substituted or unsubstituted alkyl include linear alkyl having 1 to 20 carbon atoms and branched alkyl having 3 to 20 carbon atoms.
- examples of the linear alkyl having 1 to 20 carbon atoms include the aforementioned linear alkyl having 1 to 20 carbon atoms.
- examples of the branched alkyl having 3 to 20 carbon atoms include the aforementioned branched alkyl having 3 to 20 carbon atoms.
- examples of the alkenyl in the substituted or unsubstituted alkenyl include alkenyl having 2 to 20 carbon atoms.
- examples of the alkenyl having 2 to 20 carbon atoms include the aforementioned alkenyl having 2 to 20 carbon atoms.
- examples of the substituent in the substituted alkyl and the substituted alkenyl include a phenyl group, a hydroxyl group, a sulfo group, a cyano group, and a halogen atom.
- examples of the phenyl group-substituted alkyl include benzyl and 1-phenylethyl.
- examples of the halogen atom include a chlorine atom, a bromine atom, and an iodine atom.
- Z ⁇ represents a monovalent anion.
- the halogen ion include Cl ⁇ , Br ⁇ , I ⁇ and the like.
- product (IV) examples include, for example, 1-methyl-3-octylimidazolium bromide (manufactured by Tokyo Chemical Industry), 1-methyl-3-octylimidazolium chloride (manufactured by Tokyo Chemical Industry), And dodecyl-2-methyl-3-benzylimidazolium chloride.
- an isoquinolinium salt represented by the following general formula (V) [hereinafter referred to as compound (V)] is used.
- R 11 represents substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and W ⁇ represents a monovalent anion.
- examples of the alkyl in the substituted or unsubstituted alkyl include linear alkyl having 8 to 20 carbon atoms and branched alkyl having 8 to 20 carbon atoms.
- examples of the linear alkyl having 8 to 20 carbon atoms include the aforementioned linear alkyl having 8 to 20 carbon atoms.
- examples of the branched alkyl having 8 to 20 carbon atoms include the aforementioned branched alkyl having 8 to 20 carbon atoms.
- the alkenyl in the substituted or unsubstituted alkenyl includes, for example, alkenyl having 8 to 20 carbon atoms.
- alkenyl having 8 to 20 carbon atoms include the aforementioned alkenyl having 8 to 20 carbon atoms.
- examples of the substituent in the substituted alkyl and the substituted alkenyl include a phenyl group, a hydroxyl group, a sulfo group, a cyano group, and a halogen atom.
- examples of the phenyl group-substituted alkyl include benzyl and 1-phenylethyl.
- examples of the halogen atom include a chlorine atom, a bromine atom, and an iodine atom.
- W ⁇ represents a monovalent anion.
- the monovalent anion include anions such as halogen ions.
- the halogen ion include Cl ⁇ , Br ⁇ , I ⁇ and the like.
- compound (V) examples include, for example, N-laurylisoquinolinium chloride (manufactured by NOF Corporation), N-larlylisoquinolinium bromide (manufactured by NOF Corporation) and the like.
- anionic surfactant examples include sulfate ester salt, carboxylate salt, sulfonate salt, phosphate ester salt, sulfosuccinate salt, N-methyl taurine salt, N-alkanoyl-N-methyl taurine salt and the like.
- amphoteric surfactants include tertiary amine oxides and alkylcarboxybetaines.
- nonionic surfactant examples include polyoxyethylene alkylamine, polyoxyethylene alkenylamine, polyoxyethylene alkyl ether, polyoxyethylene alkenyl ether, polyoxyethylene alkylphenyl ether, ethylenediaminetetrapolyoxyethylene, polyglycerin fatty acid.
- examples include esters, and polyoxyethylene alkylamines and polyoxyethylene alkyl ethers are preferred.
- alkyl in the polyoxyethylene alkylamine examples include alkyl having 8 to 20 carbon atoms.
- alkyl having 8 to 20 carbon atoms examples include octyl, nonyl, decyl, undecyl, dodecyl (lauryl), tridecyl, tetradecyl (myristyl), pentadecyl, hexadecyl (cetyl), heptadecyl, octadecyl (stearyl), nonadecyl, icosyl and the like. Can be mentioned.
- alkenyl in polyoxyethylene alkenylamine examples include alkenyl having 8 to 20 carbon atoms.
- alkenyl having 8 to 20 carbon atoms include octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, oleyl, nonadecenyl, icocenyl and the like.
- Examples of the alkyl in the polyoxyethylene alkyl ether include alkyl having 8 to 20 carbon atoms. Examples of the alkyl having 8 to 20 carbon atoms include the aforementioned alkyl having 8 to 20 carbon atoms. Examples of alkenyl in the polyoxyethylene alkenyl ether include alkenyl having 8 to 20 carbon atoms. Examples of the alkenyl having 8 to 20 carbon atoms include the aforementioned alkenyl having 8 to 20 carbon atoms.
- alkyl in the polyoxyethylene alkylphenyl ether examples include alkyl having 8 to 20 carbon atoms. Examples of the alkyl having 8 to 20 carbon atoms include the aforementioned alkyl having 8 to 20 carbon atoms.
- the total hemoglobin amount can be determined by a known method, for example, the cyanmethemoglobin method, the oxyhemoglobin method, the SLS-hemoglobin method, or the like.
- the total hemoglobin amount includes not only the hemoglobin-containing sample itself, but also a sample obtained by adding an isothiazolinone derivative and / or a surfactant to a hemoglobin-containing sample, an isothiazolinone derivative and / or a surfactant and a proteolytic enzyme to a hemoglobin-containing sample. It can also be determined by applying a cyan methemoglobin method, an oxyhemoglobin method, or an SLS-hemoglobin method to a sample obtained by adding s.
- the reaction in which a proteolytic enzyme is allowed to act on glycated hemoglobin in a hemoglobin-containing sample in the presence of the surfactant may be any conditions as long as the proteolytic enzyme can act on glycated hemoglobin in the presence of the surfactant. But you can do it.
- the reaction between the glycated hemoglobin in the hemoglobin-containing sample and the proteolytic enzyme is preferably performed in an aqueous medium. Examples of the aqueous medium include an aqueous medium described later. 1.
- the reaction temperature in the reaction of glycated hemoglobin in a hemoglobin-containing sample with a proteolytic enzyme is usually 10 to 50 ° C., preferably 20 to 40 ° C., and the reaction time is usually 1 minute to 3 hours. 5 minutes to 1 hour is preferred.
- the concentration of the proteolytic enzyme is not particularly limited as long as the reaction between the glycated hemoglobin in the hemoglobin-containing sample and the proteolytic enzyme proceeds, and is usually 50 to 25000 kU / L, preferably 250 to 10,000 kU / L. L.
- the proteolytic enzyme is not particularly limited as long as it is an enzyme that acts on glycated hemoglobin in a hemoglobin-containing sample and generates a glycated peptide from glycated hemoglobin.
- serine protease chymotrypsin, subtilisin, etc.
- cysteine protease papain, caspase
- Etc. aspartic protease
- pepsin, cathepsin D, etc. metalloprotease (thermolysin, etc.)
- N-terminal threonine protease glutamic acid protease and the like.
- commercially available proteolytic enzymes can also be used.
- protease P “Amano” 3G examples include protease P “Amano” 3G, protease K “Amano” (above, Amano Enzyme), actinase AS, actinase E (above).
- protease P “Amano” 3G examples include protease P “Amano” 3G, protease K “Amano” (above, Amano Enzyme), actinase AS, actinase E (above).
- Kaken Pharma Co., Ltd. examples include thermolysin (manufactured by Daiwa Kasei Co., Ltd.), Sumiteam MP (manufactured by Shin Nippon Chemical Industry Co., Ltd.) and the like.
- the concentration of the surfactant in the reaction in which the proteolytic enzyme is allowed to act is not particularly limited as long as the reaction between the glycated hemoglobin in the hemoglobin-containing sample and the proteolytic enzyme proceeds, and usually 0.0001 to 10 %, Preferably 0.0005 to 5%.
- a reaction product containing a glycated peptide is generated by a reaction between glycated hemoglobin in a hemoglobin-containing sample and a proteolytic enzyme.
- the glycated peptide in this reaction product reacts with fructosyl peptide oxidase to produce hydrogen peroxide.
- the reaction between the glycated peptide and fructosyl peptide oxidase is preferably performed in an aqueous medium. Examples of the aqueous medium include an aqueous medium described later.
- the reaction temperature in the reaction between the glycated peptide and fructosyl peptide oxidase is usually 10 to 50 ° C., preferably 20 to 40 ° C., and the reaction time is usually 1 minute to 3 hours, and 2.5 minutes to 1 Time is preferred.
- the concentration of fructosyl peptide oxidase is not particularly limited as long as the reaction between glycated hemoglobin and fructosyl peptide oxidase proceeds, and is usually 0.1 to 30 kU / L, preferably 0.2 to 15 kU / L.
- the fructosyl peptide oxidase is not particularly limited as long as it is an enzyme that acts on a glycated peptide to generate hydrogen peroxide.
- fructosyl derived from filamentous fungi, yeast, actinomycetes, bacteria, or archaea. Peptide oxidase etc. are mentioned.
- commercially available fructosyl peptide oxidase can also be used. Examples of commercially available products include FPOX-CE (manufactured by Kikkoman), FPOX-EE (manufactured by Kikkoman), and FPOX-CET (manufactured by Kikkoman). Etc.
- Examples of the method for measuring the generated hydrogen peroxide include a method using an electrode, a method using a hydrogen peroxide measuring reagent, and the like, and a method using a hydrogen peroxide measuring reagent is preferable.
- the reagent for measuring hydrogen peroxide is a reagent for converting hydrogen peroxide into a detectable substance.
- Examples of the detectable substance include a dye, light (luminescence), fluorescence, and the like, and a dye is preferable.
- examples of the reagent for measuring hydrogen peroxide include a reagent containing a peroxidase active substance such as peroxidase and an oxidative coloring type chromogen.
- examples of the oxidative coloring type chromogen include an oxidative coupling type chromogen and a leuco chromogen, and a leuco chromogen is preferred.
- leuco chromogen examples include phenothiazine chromogen, triphenylmethane chromogen, diphenylamine chromogen, o-phenylenediamine, hydroxypropionic acid, diaminobenzidine, tetramethylbenzidine and the like, and phenothiazine.
- a chromogen is preferred.
- the phenothiazine chromogen examples include 10-N-carboxymethylcarbamoyl-3,7-bis (dimethylamino) -10H-phenothiazine (CCAP), 10-N-methylcarbamoyl-3,7-bis (dimethylamino).
- -10H-phenothiazine MCDP
- 10-N- (carboxymethylaminocarbonyl) -3,7-bis (dimethylamino) -10H-phenothiazine sodium salt DA-67
- 10-N- (carboxymethylaminocarbonyl) -3,7-bis (dimethylamino) -10H-phenothiazine sodium salt DA-67
- 10-N- (carboxymethylaminocarbonyl) -3,7-bis (dimethylamino) -10H-phenothiazine sodium salt DA-67
- triphenylmethane chromogen examples include N, N, N ′, N ′, N ′′, N ′′ -hexa (3-sulfopropyl) -4,4 ′, 4 ′′ -triaminotriphenyl And methane (TPM-PS).
- diphenylamine chromogen examples include N- (carboxymethylaminocarbonyl) -4,4′-bis (dimethylamino) diphenylamine ⁇ ⁇ ⁇ ⁇ sodium salt (DA-64), 4,4′-bis (dimethylamino) diphenylamine, bis [3-bis (4-chlorophenyl) methyl-4-dimethylaminophenyl] amine (BCMA) and the like.
- examples of the reagent for measuring hydrogen peroxide include a reagent containing a peroxidase active substance such as peroxidase and a chemiluminescent substance.
- examples of the chemiluminescent substance include luminol, isoluminol, lucigenin, and acridinium ester.
- examples of the reagent for measuring hydrogen peroxide include a reagent containing a peroxidase active substance such as peroxidase and a fluorescent substance.
- examples of the fluorescent substance include 4-hydroxyphenylacetic acid, 3- (4-hydroxyphenyl) propionic acid, and coumarin.
- the glycated hemoglobin measurement reagent in a sample containing hemoglobin of the present invention is a reagent containing a proteolytic enzyme, fructosyl peptide oxidase, isothiazolinone derivative, and a surfactant.
- the method for measuring glycated hemoglobin in the hemoglobin-containing sample of the present invention may further contain a hydrogen peroxide measurement reagent.
- proteolytic enzyme fructosyl peptide oxidase, isothiazolinone derivative, surfactant, and hydrogen peroxide measuring reagent in the measurement reagent of the present invention
- proteolytic enzyme fructosyl peptide oxidase, isothiazolinone derivative, surfactant, and hydrogen peroxide measuring reagent in the measurement reagent of the present invention
- examples include derivatives, surfactants, and hydrogen peroxide measuring reagents.
- the concentration of the proteolytic enzyme in the measurement reagent of the present invention is usually 50 to 25000 kU / L, preferably 250 to 10000 kU / L.
- the concentration of fructosyl peptide oxidase in the measurement reagent of the present invention is usually 0.1 to 30 kU / L, preferably 0.2 to 15 kU / L.
- the concentration of the isothiazolinone derivative in the measurement reagent of the present invention is usually 0.005 to 20 mmol / L, preferably 0.01 to 10 mmol / L.
- the concentration of the surfactant in the measurement reagent of the present invention is usually 0.0001 to 10%, preferably 0.0005 to 5%.
- the measurement reagent of the present invention may contain an aqueous medium, a stabilizer, an antiseptic, a salt, an interference substance erasing agent, an organic solvent, and the like, if necessary.
- the aqueous medium include deionized water, distilled water, and a buffer solution, and a buffer solution is preferable.
- the pH of the aqueous medium is, for example, pH 4-10.
- a buffer solution is used as the aqueous medium, it is desirable to use a buffering agent corresponding to the set pH.
- the buffer used in the buffer include tris (hydroxymethyl) aminomethane buffer, phosphate buffer, borate buffer, Good's buffer, and the like.
- Good buffering agents include, for example, 2-morpholinoethanesulfonic acid (MES), bis (2-hydroxyethyl) iminotris (hydroxymethyl) methane (Bis-Tris), N- (2-acetamido) iminodiacetic acid (ADA) Piperazine-N, N′-bis (2-ethanesulfonic acid) (PIPES), N- (2-acetamido) -2-aminoethanesulfonic acid (ACES), 3-morpholino-2-hydroxypropanesulfonic acid (MOPSO) ), N, N-bis (2-hydroxyethyl) -2-aminoethanesulfonic acid (BES), 3-morpholinopropanesulfonic acid (MOPS), N- [tris (hydroxymethyl) methyl] -2-aminoethanesulfone Acid (TES), 2- [4- (2-hydroxyethyl) -1-piperazinyl] ethane Sulfonic acid (HEPE
- the concentration of the buffer is usually 0.001 to 2.0 mol / L, preferably 0.005 to 1.0 mol / L.
- stabilizers include polyoxyethylene surfactants such as ethylenediaminetetraacetic acid (EDTA), sucrose, calcium chloride, calcium acetate, calcium nitrate, potassium ferrocyanide, bovine serum albumin (BSA), and polyoxyethylene alkylphenyl ether. Agents and the like.
- the preservative include sodium azide and antibiotics.
- the salts include sodium chloride, sodium nitrate, sodium sulfate, sodium carbonate, sodium formate, sodium acetate, potassium chloride, potassium nitrate, potassium sulfate, potassium carbonate, potassium formate, and potassium acetate.
- the interfering substance eliminating agent include ascorbic acid oxidase for eliminating the influence of ascorbic acid.
- organic solvent examples include dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dioxane, acetone, methanol, ethanol and the like for use as a solubilizing agent for the leuco chromogen in an aqueous medium.
- Kit for measuring glycated hemoglobin in a sample containing hemoglobin The reagent for measuring glycated hemoglobin in a sample containing hemoglobin of the present invention may be stored, distributed and used in the form of a kit.
- the kit for measuring glycated hemoglobin in a hemoglobin-containing sample of the present invention is used in the method for measuring glycated hemoglobin in a hemoglobin-containing sample of the present invention.
- Examples of the measurement kit of the present invention include a two-reagent kit, a three-reagent kit, and the like, and a two-reagent kit is preferable.
- the glycated hemoglobin measurement kit in the hemoglobin-containing sample of the present invention is not particularly limited as long as it is a kit that enables the method for measuring glycated hemoglobin in the hemoglobin-containing sample of the present invention.
- a kit containing a first reagent containing a degrading enzyme and a second reagent containing a fructosyl peptide oxidase, an isothiazolinone derivative and a surfactant, and a first containing a proteolytic enzyme, an isothiazolinone derivative and a surfactant examples thereof include a kit containing a reagent and a second reagent containing fructosyl peptide oxidase.
- kits in which a hydrogen peroxide measurement reagent is contained in either or both of the first reagent and the second reagent of these kits include kits in which a hydrogen peroxide measurement reagent is contained in either or both of the first reagent and the second reagent of these kits.
- a hydrogen peroxide measurement reagent is contained in either or both of the first reagent and the second reagent of these kits.
- peroxidase and leuco chromogen are contained in separate reagents. That is, it is preferable that the peroxidase and the leuco chromogen are contained in the first reagent and the second reagent or in the second reagent and the first reagent, respectively.
- the concentration of the proteolytic enzyme in the reagent constituting the measurement kit of the present invention is usually 100 to 30000 kU / L, preferably 500 to 10000 kU / L.
- the concentration of fructosyl peptide oxidase in the reagent constituting the measurement kit of the present invention is usually 0.5 to 100 kU / L, preferably 1 to 50 kU / L.
- the concentration of the isothiazolinone derivative in the reagent constituting the measurement kit of the present invention is usually 0.005 to 20 mmol / L, preferably 0.01 to 10 mmol / L.
- the concentration of the surfactant in the reagent constituting the measurement kit of the present invention is usually 0.0001 to 40%, preferably 0.0005 to 20%.
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L C12py 1.6g / L 2-Octyl-4-isothiazolin-3-one 0.2 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L Peroxidase 40 kU / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L DA-67 60 ⁇ mol / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L C12py 1.6g / L 1,2-Benzisothiazol-3 (2H) -one 0.4 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L Peroxidase 40 kU / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L DA-67 60 ⁇ mol / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L C10TMA 16g / L 2-Octyl-4-isothiazolin-3-one 0.2 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L Peroxidase 40 kU / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L DA-67 60 ⁇ mol / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L C10TMA 16g / L 1,2-Benzisothiazol-3 (2H) -one 0.2 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L Peroxidase 40 kU / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L DA-67 60 ⁇ mol / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L LMT 5g / L 2-Octyl-4-isothiazolin-3-one 0.4 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L Peroxidase 40 kU / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L DA-67 60 ⁇ mol / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L LMT 5g / L 1,2-Benzisothiazol-3 (2H) -one 0.2 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L Peroxidase 40 kU / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L DA-67 60 ⁇ mol / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L
- Calcium chloride dihydrate 10mmol / L
- Thermolysin 1800kU / L Peroxidase 40 kU / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L DA-67 60 ⁇ mol / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L
- Calcium chloride dihydrate 10mmol / L
- Thermolysin 1800kU / L Peroxidase 40 kU / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L DA-67 60 ⁇ mol / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L C12py 1.6g / L 2-Octyl-4-isothiazolin-3-one 0.4 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L DA-67 20 ⁇ mol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L Peroxidase 120 kU / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L C12py 1.6g / L 1,2-Benzisothiazol-3 (2H) -one 0.2 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L DA-67 20 ⁇ mol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L Peroxidase 120 kU / L
- HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L C10TMA 16g / L 2-Octyl-4-isothiazolin-3-one 0.4 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L DA-67 20 ⁇ mol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L Peroxidase 120 kU / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L C10TMA 16g / L 1,2-Benzisothiazol-3 (2H) -one 0.2 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L DA-67 20 ⁇ mol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L Peroxidase 120 kU / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L LMT 5g / L 2-Octyl-4-isothiazolin-3-one 0.2 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L DA-67 20 ⁇ mol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L Peroxidase 120 kU / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L LMT 5g / L 1,2-Benzisothiazol-3 (2H) -one 0.2 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L DA-67 20 ⁇ mol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L Peroxidase 120 kU / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L Anon BL 5g / L 2-Octyl-4-isothiazolin-3-one 0.4 g / L Calcium chloride dihydrate 10mmol / L Thermolysin 1800kU / L DA-67 20 ⁇ mol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L Peroxidase 120 kU / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent Bis-Tris (pH 6.8) 10 mmol / L
- Calcium chloride dihydrate 10mmol / L
- Thermolysin 1800kU / L DA-67 20 ⁇ mol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CE 6kU / L Peroxidase 120 kU / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent MES pH 6.0
- 20 mmol / L C12py 1.2g / L 2-Octyl-4-isothiazolin-3-one 0.2 g / L
- Calcium acetate 10mmol / L Sodium nitrate 100mmol / L Actinase E 340 kU / L DA-67 20 ⁇ mol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CET 2.5 kU / L Triton X-405 7.1g / L Peroxidase 120 kU / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent MES pH 6.5
- 20 mmol / L C12py 1.6g / L 2-Octyl-4-isothiazolin-3-one 0.2 g / L Calcium nitrate 10mmol / L Sodium nitrate 100mmol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CET 6kU / L Triton X-405 7.1g / L Peroxidase 120 kU / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent MES pH 6.5
- 20 mmol / L C12py 1.6g / L 4,5-dichloro-2-octyl-4-isothiazolin-3-one 0.04 g / L
- Sodium nitrate 100mmol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CET 6kU / L Triton X-405 7.1g / L Peroxidase 120 kU / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent MES pH 6.5
- 20 mmol / L C12py 1.6g / L 2-Octyl-4-isothiazolin-3-one
- 0.2 g / L Calcium nitrate 10mmol / L Sodium nitrate 100mmol / L Actinase E 340 kU / L DA-67 20 ⁇ mol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CET 6kU / L Triton X-405 7.1g / L Peroxidase 120 kU / L
- An HbA1c measurement kit comprising the following first reagent and second reagent was prepared.
- First reagent MES pH 6.5
- 20 mmol / L C12py 1.6g / L 4,5-dichloro-2-octyl-4-isothiazolin-3-one 0.04 g / L
- Calcium nitrate 10mmol / L
- Sodium nitrate 100mmol / L Actinase E 340 kU / L DA-67 20 ⁇ mol / L
- Second reagent ADA (pH 7.0) 50 mmol / L FPOX-CET 6kU / L Triton X-405 7.1g / L Peroxidase 120 kU / L
- Example 1 Using the kit of Example 1 as a HbA1c measurement kit and whole blood derived from 10 subjects suspected of having diabetes as a sample, the total hemoglobin concentration (quantity) of HbA1c concentration (quantity) in each sample by the following procedure ) [HbA1c (%)].
- JDS Japan Diabetes Society
- each blood cell HbA1c (%) in the fraction was determined by an immunoassay using a determiner L HbA1c (manufactured by Kyowa Medex) according to the method described in the package insert of the determiner L HbA1c.
- the measurement method using the kit of Comparative Example 1 that does not contain an isothiazolinone derivative is strongly affected by hemoglobin, and accurate measurement of HbA1c is difficult, whereas the kits of Examples 1 and 2 that contain an isothiazolinone derivative are difficult. It has been found that the measurement method of the present invention using HbA1c is not affected by hemoglobin and can accurately measure HbA1c.
- reaction absorbance for each specimen was measured by the same method using the kits of Examples 9, 10 and Comparative Example 5 instead of the kit of Example 1. The results are shown in FIG.
- the measurement method using the kit of Comparative Example 5 that does not contain an isothiazolinone derivative is strongly affected by hemoglobin, and it is difficult to accurately measure HbA1c, whereas the kits of Examples 9 and 10 containing an isothiazolinone derivative are difficult. It has been found that the measurement method of the present invention using HbA1c is not affected by hemoglobin and can accurately measure HbA1c.
- Test Example 2 Effect of isothiazolinone derivative (2) As a kit, the kit of Example 1 is used as a specimen, and the hemolyzed specimen prepared in (1) of Test Example 1 is used as a specimen.
- the HbA1c concentration ( ⁇ mol / L) in each sample was determined from a calibration curve showing the relationship between the HbA1c concentration ( ⁇ mol / L) and the absorbance.
- the hemoglobin concentration in each sample was measured using hemoglobin B-Test Wako, and the hemoglobin concentration ( ⁇ mol) in each sample was determined from the obtained measurement value and the calibration curve prepared in Example 9 (1). / L) was determined.
- HbA1c concentration ( ⁇ mol / L) and hemoglobin concentration ( ⁇ mol / L) in each specimen.
- JDS Japan Diabetes Society
- kits of Examples 2 to 13, 15, 17 to 22 and Comparative Examples 1 to 11 instead of the kit of Example 1, and each sample was subjected to each sample.
- the HbA1c concentration (%) was measured.
- the HbA1c concentration (%) in the specimen having a hemoglobin concentration of 6 mg / mL was defined as a reference 0, and the difference [ ⁇ HbA1c concentration (%)] of the HbA1c concentration (%) of each specimen from the reference was calculated.
- the results are shown in Table 2.
- the ratio (%) of HbA1c to the total hemoglobin is constant regardless of the hemoglobin concentration. Therefore, the closer the ⁇ HbA1c concentration (%) is to 0, the less the HbA1c measurement method is affected by the hemoglobin concentration.
- Table 2 it was found that the kit of the present invention containing the isothiazolinone derivative was not affected by the hemoglobin concentration as compared with the kit of the comparative example not containing the isothiazolinone derivative.
- each kit of Examples 15 and 16 and Comparative Example 8 was used as a kit, and the hemolyzed sample prepared in (1) of Test Example 1 was used as a sample.
- the reaction absorbance for each specimen was measured. The results are shown in FIG.
- each kit of Examples 21 and 22 and Comparative Example 11 was used as a kit, and the hemolyzed sample prepared in (1) of Test Example 1 was used as a sample.
- the reaction absorbance for each specimen was measured. The results are shown in FIG.
- the absorbance of the reaction increased in proportion to the hemoglobin concentration in both the kit of the present invention and the kit of the comparative example, but the kit of the present invention containing the isothiazolinone derivative was used.
- the reaction absorbance was higher in the hemoglobin sample having the same concentration (that is, the sample having the same concentration of HbA1c) as compared to the case of using the kit of the comparative example not containing isothiazolinone.
- the hemolyzed specimen used for the measurement is prepared from the same human blood, the HbA1c concentration increases depending on the hemoglobin concentration.
- the higher the reaction absorbance the higher the reliability as measurement data, without being affected by hemoglobin.
- the kits of Examples 7 and 8 were used, a higher reaction absorbance was obtained in the specimen having the same concentration of HbA1c than when the kit of Comparative Example 4 was used.
- kits of Examples 15 and 16 are used, the kit of Examples 21 and 22 is compared with the kit of Comparative Example 11 as compared to the case of using the kit of Comparative Example 8.
- a high reaction absorbance was obtained in the specimen having the same concentration of HbA1c. Therefore, the measurement method using the kit of the present invention containing an isothiazolinone derivative is less affected by hemoglobin and has a higher sensitivity for the measurement of HbA1c than the measurement method using a comparative kit containing no isothiazolinone derivative. It turned out to be possible.
- a measurement method, a measurement reagent, and a measurement kit for glycated hemoglobin in a hemoglobin-containing sample which are useful for measurement of glycated hemoglobin useful for diagnosis of diabetes.
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
[2] イソチアゾリノン誘導体が、下記式(I)で表わされる化合物である[1]記載の測定方法。
[3] 式(I)で表わされる化合物が、2-アルキル-4-イソチアゾリン-3-オン、1,2-ベンゾイソチアゾール-3(2H)-オン、及び、2-アルキル-4,5-ジハロゲノ-4-イソチアゾリン-3-オンからなる群より選ばれるイソチアゾリノン誘導体である[2]記載の測定方法。
[4] 過酸化水素の測定が、過酸化水素測定試薬により行われる、[1]~[3]のいずれかに記載の測定方法。
[5] 過酸化水素測定試薬が、ペルオキシダーゼとロイコ型色原体とを含む試薬である[4]記載の測定方法。
[7] イソチアゾリノン誘導体が、下記式(I)で表わされる化合物である[6]記載の測定試薬。
[8] 式(I)で表わされる化合物が、2-アルキル-4-イソチアゾリン-3-オン、1,2-ベンゾイソチアゾール-3(2H)-オン、及び、2-アルキル-4,5-ジハロゲノ-4-イソチアゾリン-3-オンからなる群より選ばれるイソチアゾリノン誘導体である[7]記載の測定試薬。
[9] さらに、過酸化水素測定試薬を含む、[6]~[8]のいずれかに記載の測定試薬。
[10] 過酸化水素測定試薬が、ペルオキシダーゼとロイコ型色原体とを含む試薬である[9]記載の試薬。
[12] ヘモグロビン含有試料中の糖化ヘモグロビンを測定するためのキットであって、タンパク質分解酵素、及び、界面活性剤を含む第1試薬と、フルクトシルペプチド酸化酵素、及び、イソチアゾリノン誘導体を含む第2試薬とを含むことを特徴とする糖化ヘモグロビン測定キット。
[13] イソチアゾリノン誘導体が、下記式(I)で表わされる化合物である[11]又は[12]記載の測定キット。
[14] 式(I)で表わされる化合物が、2-アルキル-4-イソチアゾリン-3-オン、1,2-ベンゾイソチアゾール-3(2H)-オン、及び、2-アルキル-4,5-ジハロゲノ-4-イソチアゾリン-3-オンからなる群より選ばれるイソチアゾリノン誘導体である[13]記載の測定キット。
[15] さらに、ペルオキシダーゼ及びロイコ型色原体が、それぞれ第1試薬と第2試薬、又は、第2試薬と第1試薬に含まれる、[11]~[14]のいずれかに記載の測定キット。
本発明の、ヘモグロビン含有試料中の糖化ヘモグロビンの測定方法は、ヘモグロビン含有試料中の糖化ヘモグロビンに、界面活性剤存在下に、タンパク質分解酵素を作用させた後、フルクトシルペプチド酸化酵素を作用させる反応において、当該反応をイソチアゾリノン誘導体存在下に行い、生成する過酸化水素を測定する方法である。イソチアゾリノン誘導体は、フルクトシルペプチド酸化酵素を作用させる反応に存在させても、タンパク質分解酵素を作用させる反応とフルクトシルペプチド酸化酵素を作用させる反応の両方の反応に存在させてもよい。
具体的には、以下の工程を含む測定方法が例示される。
(1)ヘモグロビン含有試料中の糖化ヘモグロビンに、界面活性剤存在下に、タンパク質分解酵素を作用させる工程;
(2)工程(1)で得られる反応生成物に、イソチアゾリノン誘導体存在下に、フルクトシルペプチド酸化酵素を作用させ、過酸化水素を生成させる工程;
(3)工程(2)で生成した過酸化水素を測定する工程;及び、
(4)既知濃度の糖化ヘモグロビンを用いて予め作成した、過酸化水素量と糖化ヘモグロビン濃度との関係を表す検量線に基づき、工程(3)で測定した過酸化水素量から、ヘモグロビン含有試料中の糖化ヘモグロビン濃度を決定する工程。
(1)ヘモグロビン含有試料中の糖化ヘモグロビンに、イソチアゾリノン誘導体及び界面活性剤の存在下に、タンパク質分解酵素を作用させる工程;
(2)工程(1)で得られる反応生成物に、フルクトシルペプチド酸化酵素と反応させ、過酸化水素を生成させる工程;
(3)工程(2)で生成した過酸化水素を測定する工程;及び、
(4)既知濃度の糖化ヘモグロビンを用いて予め作成した、過酸化水素量と糖化ヘモグロビン濃度との関係を表す検量線に基づき、工程(3)で測定した過酸化水素量から、ヘモグロビン含有試料中の糖化ヘモグロビン濃度を決定する工程。
(1)ヘモグロビン含有試料中の総ヘモグロビン(すなわち、ヘモグロビンと糖化ヘモグロビンを合わせた総ヘモグロビン)量を決定する工程;
(2)ヘモグロビン含有試料中の糖化ヘモグロビンに、界面活性剤存在下、タンパク質分解酵素を作用させる工程;
(3)工程(2)で得られる反応生成物に、イソチアゾリノン誘導体存在下に、フルクトシルペプチド酸化酵素を作用させ、過酸化水素を生成させる工程;
(4)工程(3)で生成した過酸化水素を測定する工程;及び、
(5)既知量の糖化ヘモグロビンを用いて予め作成した、過酸化水素量と糖化ヘモグロビン量との関係を表す検量線に基づき、工程(4)で測定した過酸化水素量から、ヘモグロビン含有試料中の糖化ヘモグロビン量を決定する工程;及び、
(6)工程(1)で決定した総ヘモグロビン量と、工程(5)で決定した糖化ヘモグロビン量から、ヘモグロビン含有試料中の糖化ヘモグロビン量の総ヘモグロビン量に対する割合を算出する工程。
上記工程(1)の総ヘモグロビン量の決定は、工程(2)の後に行うこともできる。
(1)ヘモグロビン含有試料中の総ヘモグロビン(すなわち、ヘモグロビンと糖化ヘモグロビンを合わせた総ヘモグロビン)量を決定する工程;
(2)ヘモグロビン含有試料中の糖化ヘモグロビンに、イソチアゾリノン誘導体と界面活性剤存在下、タンパク質分解酵素を作用させる工程;
(3)工程(2)で得られる反応生成物に、フルクトシルペプチド酸化酵素を作用させ、過酸化水素を生成させる工程;
(4)工程(3)で生成した過酸化水素を測定する工程;及び、
(5)既知量の糖化ヘモグロビンを用いて予め作成した、過酸化水素量と糖化ヘモグロビン量との関係を表す検量線に基づき、工程(4)で測定した過酸化水素量から、ヘモグロビン含有試料中の糖化ヘモグロビン量を決定する工程;及び、
(6)工程(1)で決定した総ヘモグロビン量と、工程(5)で決定した糖化ヘモグロビン量から、ヘモグロビン含有試料中の糖化ヘモグロビン量の総ヘモグロビン量に対する割合を算出する工程。
上記工程(1)の総ヘモグロビン量の決定は、工程(2)の後に行うこともできる。
ポリオキシエチレンアルケニルエーテルにおけるアルケニルとしては、例えば炭素数8~20のアルケニル等が挙げられる。炭素数8~20のアルケニルとしては、例えば前述の炭素数8~20のアルケニル等が挙げられる。
本発明の、ヘモグロビン含有試料中の糖化ヘモグロビン測定試薬は、タンパク質分解酵素、フルクトシルペプチド酸化酵素、イソチアゾリノン誘導体、及び、界面活性剤を含む試薬であり、本発明のヘモグロビン含有試料中の糖化ヘモグロビンの測定方法に用いられる。本発明の測定試薬は、さらに、過酸化水素測定試薬を含んでいてもよい。
水性媒体としては、例えば脱イオン水、蒸留水、緩衝液等が挙げられ、緩衝液が好ましい。
本発明のヘモグロビン含有試料中の糖化ヘモグロビン測定試薬は、キットの形態で保存、流通及び使用されてよい。本発明のヘモグロビン含有試料中の糖化ヘモグロビン測定キットは、本発明のヘモグロビン含有試料中の糖化ヘモグロビンの測定方法に用いられる。本発明の測定キットとしては、例えば2試薬系のキット、3試薬系のキット等が挙げられ、2試薬系キットが好ましい。
本発明の測定キットを構成する試薬中のイソチアゾリノン誘導体の濃度は、通常0.005~20mmol/Lであり、好ましくは0.01~10mmol/Lである。
尚、本実施例、比較例及び試験例においては、下記メーカーの試薬及び酵素を使用した。
第1試薬
Bis-Tris(pH6.8) 10mmol/L
C12py 1.6g/L
2-オクチル-4-イソチアゾリン-3-オン 0.2g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
ペルオキシダーゼ 40kU/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
DA-67 60μmol/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
C12py 1.6g/L
1,2-ベンゾイソチアゾール-3(2H)-オン 0.4g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
ペルオキシダーゼ 40kU/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
DA-67 60μmol/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
C10TMA 16g/L
2-オクチル-4-イソチアゾリン-3-オン 0.2g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
ペルオキシダーゼ 40kU/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
DA-67 60μmol/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
C10TMA 16g/L
1,2-ベンゾイソチアゾール-3(2H)-オン 0.2g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
ペルオキシダーゼ 40kU/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
DA-67 60μmol/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
LMT 5g/L
2-オクチル-4-イソチアゾリン-3-オン 0.4g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
ペルオキシダーゼ 40kU/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
DA-67 60μmol/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
LMT 5g/L
1,2-ベンゾイソチアゾール-3(2H)-オン 0.2g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
ペルオキシダーゼ 40kU/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
DA-67 60μmol/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
アノンBL 5g/L
2-オクチル-4-イソチアゾリン-3-オン 0.2g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
ペルオキシダーゼ 40kU/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
DA-67 60μmol/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
アノンBL 5g/L
1,2-ベンゾイソチアゾール-3(2H)-オン 0.4g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
ペルオキシダーゼ 40kU/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
DA-67 60μmol/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
C12py 1.6g/L
2-オクチル-4-イソチアゾリン-3-オン 0.4g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
ペルオキシダーゼ 120kU/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
C12py 1.6g/L
1,2-ベンゾイソチアゾール-3(2H)-オン 0.2g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
ペルオキシダーゼ 120kU/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
C10TMA 16g/L
2-オクチル-4-イソチアゾリン-3-オン 0.4g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
ペルオキシダーゼ 120kU/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
C10TMA 16g/L
1,2-ベンゾイソチアゾール-3(2H)-オン 0.2g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
ペルオキシダーゼ 120kU/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
LMT 5g/L
2-オクチル-4-イソチアゾリン-3-オン 0.2g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
ペルオキシダーゼ 120kU/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
LMT 5g/L
1,2-ベンゾイソチアゾール-3(2H)-オン 0.2g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
ペルオキシダーゼ 120kU/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
アノンBL 5g/L
2-オクチル-4-イソチアゾリン-3-オン 0.4g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
ペルオキシダーゼ 120kU/L
第1試薬
Bis-Tris(pH6.8) 10mmol/L
アノンBL 5g/L
1,2-ベンゾイソチアゾール-3(2H)-オン 0.4g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
ペルオキシダーゼ 120kU/L
第1試薬
MES(pH6.0) 20mmol/L
C12py 1.2g/L
2-オクチル-4-イソチアゾリン-3-オン 0.2g/L
酢酸カルシウム 10mmol/L
硝酸ナトリウム 100mmol/L
アクチナーゼE 340kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CET 2.5kU/L
トリトンX-405 7.1g/L
ペルオキシダーゼ 120kU/L
第1試薬
MES(pH6.0) 20mmol/L
C12py 1.2g/L
1,2-ベンゾイソチアゾール-3(2H)-オン 0.04g/L
酢酸カルシウム 10mmol/L
硝酸ナトリウム 100mmol/L
アクチナーゼE 340kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CET 2.5kU/L
トリトンX-405 7.1g/L
ペルオキシダーゼ 120kU/L
第1試薬
MES(pH6.5) 20mmol/L
C12py 1.6g/L
2-オクチル-4-イソチアゾリン-3-オン 0.2g/L
硝酸カルシウム 10mmol/L
硝酸ナトリウム 100mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CET 6kU/L
トリトンX-405 7.1g/L
ペルオキシダーゼ 120kU/L
第1試薬
MES(pH6.5) 20mmol/L
C12py 1.6g/L
4,5-ジクロロ-2-オクチル-4-イソチアゾリン-3-オン
0.04g/L
硝酸カルシウム 10mmol/L
硝酸ナトリウム 100mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CET 6kU/L
トリトンX-405 7.1g/L
ペルオキシダーゼ 120kU/L
第1試薬
MES(pH6.5) 20mmol/L
C12py 1.6g/L
2-オクチル-4-イソチアゾリン-3-オン 0.2g/L
硝酸カルシウム 10mmol/L
硝酸ナトリウム 100mmol/L
アクチナーゼE 340kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CET 6kU/L
トリトンX-405 7.1g/L
ペルオキシダーゼ 120kU/L
第1試薬
MES(pH6.5) 20mmol/L
C12py 1.6g/L
4,5-ジクロロ-2-オクチル-4-イソチアゾリン-3-オン
0.04g/L
硝酸カルシウム 10mmol/L
硝酸ナトリウム 100mmol/L
アクチナーゼE 340kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CET 6kU/L
トリトンX-405 7.1g/L
ペルオキシダーゼ 120kU/L
測定キットとして、総ヘモグロビン測定用キットであるヘモグロビンB-テストワコー(SLS-ヘモグロビン法)(和光純薬工業社製)を用いて、検体として、ヘモグロビンB-テストワコーに付属の標準品(ヘモグロビン濃度:15.3mg/mL)を用いて、ヘモグロビン濃度と吸光度との関係を示す検量線を作成した。
ラテックス免疫凝集法と、血球画分の総ヘモグロビン値とから、HbA1c濃度が2.77μmol/L、6.33μmol/Lと値付けされた2つの血球画分について、実施例1のHbA1c測定キットを用いて測定し、各血球画分に対する吸光度を測定した。当該血球画分の代わりに生理食塩水を用いて、生理食塩水に対するHbA1c濃度を測定した。当該血球画分に対するそれぞれの吸光度から、生理食塩水に対する吸光度を差し引いて算出した値を、当該血球画分に対するブランク補正吸光度とした。当該血球画分に対するブランク補正吸光度と、生理食塩水に対するブランク補正吸光度(0 Abs)とから、HbA1c濃度(μmol/L)と吸光度との間の関係を示す検量線を作成した。
各試料に対して、25℃、3000rpmで5分間遠心分離を行い、血球画分を得た。各血球画分について、ヘモグロビンB-テストワコーを用いて測定し、得られた測定値と(1)の検量線とから、各血球画分中のヘモグロビン濃度(μmol/L)を決定した。
各血球画分について、実施例1の測定キットを用いて測定し、得られた測定値と(2)の検量線とから、各血球画分中のHbA1c濃度(μmol/L)を決定した。
上記(3)で決定した各血球画分におけるヘモグロビン濃度(μmol/L)と、上記(4)で決定した各血球画分におけるHbA1c濃度(μmol/L)とから、以下の式により、日本糖尿病学会(Japan Diabetes Society;JDS)値のHbA1c(%)を算出した。
上記(5)でのHbA1c(%)の決定に使用した血球画分と同一の血球画分を用いて、各血球画分中のHbA1c(%)を、デタミナーL HbA1c(協和メデックス社製)を用いる免疫測定法により、デタミナーL HbA1cの添付文書に記載の方法に従って決定した。
本発明の測定方法を用いて、上記(5)で決定したHbA1c(%)と、免疫測定法を用いて、上記(6)で決定したHbA1c(%)とから、本発明の測定方法と免疫測定法との間の相関関係を検証し、相関係数を決定した。
以下の第1試薬及び第2試薬からなるHbA1c測定キットを調製した。
第1試薬
Bis-Tris(pH6.8) 10mmol/L
C12py 1.6g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
ペルオキシダーゼ 40kU/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
DA-67 60μmol/L
以下の第1試薬及び第2試薬からなるHbA1c測定キットを調製した。
第1試薬
Bis-Tris(pH6.8) 10mmol/L
C10TMA 16g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
ペルオキシダーゼ 40kU/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
DA-67 60μmol/L
以下の第1試薬及び第2試薬からなるHbA1c測定キットを調製した。
第1試薬
Bis-Tris(pH6.8) 10mmol/L
LMT 5g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
ペルオキシダーゼ 40kU/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
DA-67 60μmol/L
以下の第1試薬及び第2試薬からなるHbA1c測定キットを調製した。
第1試薬
Bis-Tris(pH6.8) 10mmol/L
アノンBL 5g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
ペルオキシダーゼ 40kU/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
DA-67 60μmol/L
以下の第1試薬及び第2試薬からなるHbA1c測定キットを調製した。
第1試薬
Bis-Tris(pH6.8) 10mmol/L
C12py 1.6g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
ペルオキシダーゼ 120kU/L
以下の第1試薬及び第2試薬からなるHbA1c測定キットを調製した。
第1試薬
Bis-Tris(pH6.8) 10mmol/L
C10TMA 16g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
ペルオキシダーゼ 120kU/L
以下の第1試薬及び第2試薬からなるHbA1c測定キットを調製した。
第1試薬
Bis-Tris(pH6.8) 10mmol/L
LMT 5g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
ペルオキシダーゼ 120kU/L
以下の第1試薬及び第2試薬からなるHbA1c測定キットを調製した。
第1試薬
Bis-Tris(pH6.8) 10mmol/L
アノンBL 5g/L
塩化カルシウム2水和物 10mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CE 6kU/L
ペルオキシダーゼ 120kU/L
以下の第1試薬及び第2試薬からなるHbA1c測定キットを調製した。
第1試薬
MES(pH6.0) 20mmol/L
C12py 1.2g/L
酢酸カルシウム 10mmol/L
硝酸ナトリウム 100mmol/L
アクチナーゼE 340kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CET 2.5kU/L
トリトンX-405 7.1g/L
ペルオキシダーゼ 120kU/L
以下の第1試薬及び第2試薬からなるHbA1c測定キットを調製した。
第1試薬
MES(pH6.5) 20mmol/L
C12py 1.6g/L
硝酸カルシウム 10mmol/L
硝酸ナトリウム 100mmol/L
サーモリシン 1800kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CET 6kU/L
トリトンX-405 7.1g/L
ペルオキシダーゼ 120kU/L
以下の第1試薬及び第2試薬からなるHbA1c測定キットを調製した。
第1試薬
MES(pH6.5) 20mmol/L
C12py 1.6g/L
硝酸カルシウム 10mmol/L
硝酸ナトリウム 100mmol/L
アクチナーゼE 340kU/L
DA-67 20μmol/L
第2試薬
ADA(pH7.0) 50mmol/L
FPOX-CET 6kU/L
トリトンX-405 7.1g/L
ペルオキシダーゼ 120kU/L
(1)溶血検体の調製
ヒト血液を遠心分離して得られた血球画分を、ヘモグロビン測定試薬であるネスコート ヘモキット-N(アルフレッサファーマ社製)を用いて測定し、該血球画分のヘモグロビン濃度を決定した。次いで、この値付けされた該血球画分を精製水で希釈して溶血させ、ヘモグロビン濃度が2mg/mL、4mg/mL、6mg/mL、8mg/mL及び10mg/mLの各溶血検体を調製した。
キットとして、実施例1のキットを用いて、検体として、生理食塩水(ヘモグロビン濃度が0mg/mL)及び(1)で調製した溶血検体を用いて、以下の方法により、各検体に対する反応吸光度を測定した。
キットとして、実施例1のキットを、検体として、試験例1の(1)で調製した溶血検体を用いて測定を行い、得られた測定値を基に、実施例9の(2)で作成したHbA1c濃度(μmol/L)と吸光度との間の関係を示す検量線から、各検体中のHbA1c濃度(μmol/L)を決定した。一方、各検体中のヘモグロビン濃度を、ヘモグロビンB-テストワコーを用いて測定し、得られた測定値と実施例9の(1)で作成した検量線とから、各検体中のヘモグロビン濃度(μmol/L)を決定した。
キットとして、実施例7,8及び比較例4の各キットを、検体として、試験例1の(1)で調製した溶血検体を用いて、試験例1と同様の方法により、それぞれのキットにおいて、各検体に対する反応吸光度を測定した。その結果を第3図に示す。
Claims (15)
- ヘモグロビン含有試料に、界面活性剤存在下に、タンパク質分解酵素を作用させた後、フルクトシルペプチド酸化酵素を作用させる反応において、当該反応をイソチアゾリノン誘導体存在下に行い、生成した過酸化水素を測定することを特徴とする、ヘモグロビン含有試料中の糖化ヘモグロビンの測定方法。
- 式(I)で表わされる化合物が、2-アルキル-4-イソチアゾリン-3-オン、1,2-ベンゾイソチアゾール-3(2H)-オン、及び、2-アルキル-4,5-ジハロゲノ-4-イソチアゾリン-3-オンからなる群より選ばれるイソチアゾリノン誘導体である請求項2記載の測定方法。
- 過酸化水素の測定が、過酸化水素測定試薬により行われる、請求項1~3のいずれかに記載の測定方法。
- 過酸化水素測定試薬が、ペルオキシダーゼとロイコ型色原体とを含む試薬である請求項4記載の測定方法。
- ヘモグロビン含有試料中の糖化ヘモグロビンを測定するための試薬であって、タンパク質分解酵素、フルクトシルペプチド酸化酵素、イソチアゾリノン誘導体、及び、界面活性剤を含むことを特徴とする糖化ヘモグロビン測定試薬。
- 式(I)で表わされる化合物が、2-アルキル-4-イソチアゾリン-3-オン、1,2-ベンゾイソチアゾール-3(2H)-オン、及び、2-アルキル-4,5-ジハロゲノ-4-イソチアゾリン-3-オンからなる群より選ばれるイソチアゾリノン誘導体である請求項7記載の測定試薬。
- さらに、過酸化水素測定試薬を含む、請求項6~8のいずれかに記載の測定試薬。
- 過酸化水素測定試薬が、ペルオキシダーゼとロイコ型色原体とを含む試薬である請求項9記載の試薬。
- ヘモグロビン含有試料中の糖化ヘモグロビンを測定するためのキットであって、タンパク質分解酵素、イソチアゾリノン誘導体、及び、界面活性剤を含む第1試薬と、フルクトシルペプチド酸化酵素を含む第2試薬とを含むことを特徴とする糖化ヘモグロビン測定キット。
- ヘモグロビン含有試料中の糖化ヘモグロビンを測定するためのキットであって、タンパク質分解酵素、及び、界面活性剤を含む第1試薬と、フルクトシルペプチド酸化酵素、及び、イソチアゾリノン誘導体を含む第2試薬とを含むことを特徴とする糖化ヘモグロビン測定キット。
- 式(I)で表わされる化合物が、2-アルキル-4-イソチアゾリン-3-オン、1,2-ベンゾイソチアゾール-3(2H)-オン、及び、2-アルキル-4,5-ジハロゲノ-4-イソチアゾリン-3-オンからなる群より選ばれるイソチアゾリノン誘導体である請求項13記載の測定キット。
- さらに、ペルオキシダーゼ及びロイコ型色原体が、それぞれ第1試薬と第2試薬、又は、第2試薬と第1試薬に含まれる、請求項11~14のいずれかに記載の測定キット。
Priority Applications (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP11816408.6A EP2604698A4 (en) | 2010-08-11 | 2011-08-09 | METHOD FOR MEASURING GLYCED HEMOGLOBIN |
KR1020137002549A KR20130107268A (ko) | 2010-08-11 | 2011-08-09 | 당화 헤모글로빈의 측정 방법 |
JP2012528677A JP5871800B2 (ja) | 2010-08-11 | 2011-08-09 | 糖化ヘモグロビンの測定方法 |
CA2806261A CA2806261A1 (en) | 2010-08-11 | 2011-08-09 | Method for measuring glycated hemoglobin |
US13/811,911 US20130171676A1 (en) | 2010-08-11 | 2011-08-09 | Method for measuring glycated hemoglobin |
BR112013001398A BR112013001398A2 (pt) | 2010-08-11 | 2011-08-09 | método, reagente e kit para medir hemoglobina glicada |
CN201180038629.XA CN103124793B (zh) | 2010-08-11 | 2011-08-09 | 糖化血红蛋白的测定方法 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2010180562 | 2010-08-11 | ||
JP2010-180562 | 2010-08-11 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2012020744A1 true WO2012020744A1 (ja) | 2012-02-16 |
Family
ID=45567707
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2011/068103 WO2012020744A1 (ja) | 2010-08-11 | 2011-08-09 | 糖化ヘモグロビンの測定方法 |
Country Status (8)
Country | Link |
---|---|
US (1) | US20130171676A1 (ja) |
EP (1) | EP2604698A4 (ja) |
JP (1) | JP5871800B2 (ja) |
KR (1) | KR20130107268A (ja) |
CN (1) | CN103124793B (ja) |
BR (1) | BR112013001398A2 (ja) |
CA (1) | CA2806261A1 (ja) |
WO (1) | WO2012020744A1 (ja) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2013118743A1 (ja) * | 2012-02-09 | 2013-08-15 | 協和メデックス株式会社 | アスコルビン酸の影響抑制方法 |
WO2015020200A1 (ja) | 2013-08-09 | 2015-02-12 | キッコーマン株式会社 | 改変型アマドリアーゼ及びその製造法、並びにアマドリアーゼの界面活性剤耐性向上剤及びこれを用いたHbA1c測定用組成物 |
US11198852B2 (en) | 2014-11-07 | 2021-12-14 | Kikkoman Corporation | Amadoriase having enhanced anionic surfactant tolerance |
JP2021535380A (ja) * | 2018-08-22 | 2021-12-16 | クエスト ダイアグノスティクス インベストメンツ エルエルシー | 糖尿病におけるマイクロサンプリング検出 |
Families Citing this family (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105431545B (zh) * | 2013-07-09 | 2021-06-18 | 日立化成诊断系统株式会社 | 糖化血红蛋白的测定方法 |
KR102228913B1 (ko) * | 2013-12-10 | 2021-03-17 | 삼성전자주식회사 | 미세유동장치 및 검사장치 |
EP3140661B1 (de) | 2014-05-06 | 2018-08-15 | DiaSys Diagnostic Systems GmbH | Enzymatische bestimmung von hba1c |
WO2016186521A1 (en) * | 2015-05-15 | 2016-11-24 | Canterbury Scientific Limited | Haemolysis stabilising composition |
EP3492600B1 (en) * | 2016-07-29 | 2024-04-24 | Hitachi Chemical Diagnostics Systems Co., Ltd. | Method for measuring glycated hemoglobin |
CN110520728B (zh) * | 2017-03-03 | 2022-12-06 | 聚合物技术系统公司 | 用于酶促a1c检测和定量的系统和方法 |
US11703513B2 (en) | 2017-08-07 | 2023-07-18 | Polymer Technology Systems, Inc. | Systems and methods for enzymatic A1C detection and quantification |
MX2019010428A (es) * | 2017-12-12 | 2020-01-09 | Polymer Technology Systems Inc | Sistemas y métodos para la preparación de muestras para la detección y cuantificación enzimáticas de a1c. |
WO2020113129A1 (en) * | 2018-11-29 | 2020-06-04 | Polymer Technology Systems, Inc. | Systems and methods for electrochemical point-of-care detection of hemoglobin |
CN110297048A (zh) * | 2019-07-12 | 2019-10-01 | 上海交通大学 | 一种抗干扰的糖化血红蛋白同系物相对含量的测定方法 |
CN110687306B (zh) * | 2019-10-30 | 2023-04-25 | 深圳上泰生物工程有限公司 | 一种直接机上溶血酶法双试剂糖化血红蛋白检测试剂盒 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0310696A (ja) | 1989-06-09 | 1991-01-18 | Wako Pure Chem Ind Ltd | 体液成分の測定方法 |
JP2000210100A (ja) | 1998-11-17 | 2000-08-02 | Kdk Corp | 酸化還元反応を用いた測定方法 |
WO2003107011A1 (ja) | 2002-06-14 | 2003-12-24 | アークレイ株式会社 | スルホン酸化合物およびニトロ化合物を用いた測定方法 |
WO2005049858A1 (ja) | 2003-11-19 | 2005-06-02 | Daiichi Pure Chemicals Co., Ltd. | ヘモグロビン含有試料中の基質の測定方法 |
JP2007147630A (ja) | 2002-06-14 | 2007-06-14 | Arkray Inc | スルホン酸化合物およびニトロ化合物を用いた測定方法 |
Family Cites Families (22)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB8530188D0 (en) * | 1985-12-06 | 1986-01-15 | Unilever Plc | Enzymatic liquid detergent composition |
US6174728B1 (en) * | 1998-04-03 | 2001-01-16 | Avl Medical Instruments Ag | Control or calibration standard for use with instruments for optical measurement of hemoglobin concentration in blood samples |
US20020098589A1 (en) * | 2001-01-19 | 2002-07-25 | Crews Harold Richardson | Multi-purpose reagent system and method for enumeration of red blood cells, white blood cells and thrombocytes and differential determination of white blood cells |
JP3949854B2 (ja) * | 1999-10-01 | 2007-07-25 | キッコーマン株式会社 | 糖化蛋白質の測定方法 |
US7794966B2 (en) * | 2003-12-12 | 2010-09-14 | Arkray, Inc. | Method of measuring glycated amine |
EP1725866A4 (en) * | 2004-03-16 | 2010-09-29 | Fujifilm Corp | TEST CHIP |
US20070178547A1 (en) * | 2004-03-17 | 2007-08-02 | Daiichi Pure Chemicals Co., Ltd. | Method of measuring glycated protein |
JP5131955B2 (ja) * | 2004-08-05 | 2013-01-30 | 旭化成ファーマ株式会社 | プロテアーゼ反応促進剤及び/又は色素の安定化剤を含む試薬 |
US20080193482A1 (en) * | 2004-09-21 | 2008-08-14 | Woodward John R | Method of cancer screening; method of cancer treatment; and method of auto-immune disease treatment |
WO2006063009A2 (en) * | 2004-12-07 | 2006-06-15 | Ohio University | Diagnosis of hyperinsulinemia and type ii diabetes and protection against same based on proteins differentially expressed in serum |
US7541190B2 (en) * | 2005-02-07 | 2009-06-02 | Beckman Coulter, Inc. | Method of measurement of cellular hemoglobin |
US8062901B2 (en) * | 2005-04-30 | 2011-11-22 | Alere Switzerland Gmbh | Devices and methods for sample collection and analysis |
US20090117660A1 (en) * | 2005-04-30 | 2009-05-07 | Oakville Hong Kong Co., Limited | Devices and methods for sample collection and analysis |
KR101017031B1 (ko) * | 2006-04-03 | 2011-02-23 | 베이징 티안큉 케미컬스 컴퍼니 리미티드 | N-치환 이소치아졸리논 유도체의 제조 방법 |
JP5156738B2 (ja) * | 2006-06-06 | 2013-03-06 | エフ.ホフマン−ラ ロシュ アーゲー | 全血試料の特異的溶血 |
US7855079B2 (en) * | 2006-07-25 | 2010-12-21 | General Atomics | Methods for assaying percentage of glycated hemoglobin |
US7521244B2 (en) * | 2006-10-26 | 2009-04-21 | Bionostics, Inc. | Standard reference solutions |
US20090186819A1 (en) * | 2007-12-11 | 2009-07-23 | Marieve Carrier | Formulation of insulinotropic peptide conjugates |
US7968279B2 (en) * | 2008-08-13 | 2011-06-28 | Beckman Coulter, Inc. | Reference control for cell by cell analysis |
WO2010148130A1 (en) * | 2009-06-17 | 2010-12-23 | Maine Standards Company, Llc | Method for measuring lipoprotein-specific apolipoproteins |
US20130303466A1 (en) * | 2010-10-19 | 2013-11-14 | Elcelyx Therapeutics, Inc. | Chemosensory Receptor Ligand-Based Therapies |
JP2014505011A (ja) * | 2010-10-19 | 2014-02-27 | エルセリクス セラピューティクス インコーポレイテッド | 化学感覚受容体リガンドに基づく治療法 |
-
2011
- 2011-08-09 JP JP2012528677A patent/JP5871800B2/ja active Active
- 2011-08-09 BR BR112013001398A patent/BR112013001398A2/pt not_active IP Right Cessation
- 2011-08-09 US US13/811,911 patent/US20130171676A1/en not_active Abandoned
- 2011-08-09 CN CN201180038629.XA patent/CN103124793B/zh not_active Expired - Fee Related
- 2011-08-09 CA CA2806261A patent/CA2806261A1/en not_active Abandoned
- 2011-08-09 EP EP11816408.6A patent/EP2604698A4/en not_active Withdrawn
- 2011-08-09 KR KR1020137002549A patent/KR20130107268A/ko not_active Application Discontinuation
- 2011-08-09 WO PCT/JP2011/068103 patent/WO2012020744A1/ja active Application Filing
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0310696A (ja) | 1989-06-09 | 1991-01-18 | Wako Pure Chem Ind Ltd | 体液成分の測定方法 |
JP2000210100A (ja) | 1998-11-17 | 2000-08-02 | Kdk Corp | 酸化還元反応を用いた測定方法 |
WO2003107011A1 (ja) | 2002-06-14 | 2003-12-24 | アークレイ株式会社 | スルホン酸化合物およびニトロ化合物を用いた測定方法 |
JP2007147630A (ja) | 2002-06-14 | 2007-06-14 | Arkray Inc | スルホン酸化合物およびニトロ化合物を用いた測定方法 |
WO2005049858A1 (ja) | 2003-11-19 | 2005-06-02 | Daiichi Pure Chemicals Co., Ltd. | ヘモグロビン含有試料中の基質の測定方法 |
Non-Patent Citations (5)
Title |
---|
CHEW, A.L. ET AL.: "1,2-Benzisothiazolin-3-one (Proxel): irritant or allergen? A clinical study and literature review.", CONTACT DERMATITIS, vol. 36, no. 3, March 1997 (1997-03-01), pages 131 - 136, XP055070562 * |
HIROKAWA, K. ET AL.: "An enzymatic method for the determination of hemoglobinAlC.", BIOTECHNOL. LETT., vol. 27, no. 14, July 2005 (2005-07-01), pages 963 - 968, XP019230894 * |
KOZO HIROKAWA: "Development of novel enzymes applied to clinical diagnosis for diabetes", BIO INDUSTRY, vol. 26, no. 1, 12 January 2009 (2009-01-12), pages 73 - 80, XP009175348 * |
See also references of EP2604698A4 |
THORMANN, J.: "Contact dermatitis to a new fungicide, 2-n-octyl-4-isothiazolin-3-one.", CONTACT DERMATITIS, vol. 8, no. 3, May 1982 (1982-05-01), pages 204, XP055070551 * |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2013118743A1 (ja) * | 2012-02-09 | 2013-08-15 | 協和メデックス株式会社 | アスコルビン酸の影響抑制方法 |
JPWO2013118743A1 (ja) * | 2012-02-09 | 2015-05-11 | 協和メデックス株式会社 | アスコルビン酸の影響抑制方法 |
WO2015020200A1 (ja) | 2013-08-09 | 2015-02-12 | キッコーマン株式会社 | 改変型アマドリアーゼ及びその製造法、並びにアマドリアーゼの界面活性剤耐性向上剤及びこれを用いたHbA1c測定用組成物 |
US10619183B2 (en) | 2013-08-09 | 2020-04-14 | Kikkoman Corporation | Modified amadoriase and method for producing the same, agent for improving surfactant resistance of amadoriase and composition for measuring HbA1c using the same |
EP3760717A1 (en) | 2013-08-09 | 2021-01-06 | Kikkoman Corporation | Amadoriase and method for producing the same, agent for improving surfactant resistance of amadoriase and composition for measuring hba1c using the same |
US11549134B2 (en) | 2013-08-09 | 2023-01-10 | Kikkoman Corporation | Modified amadoriase and method for producing the same, agent for improving surfactant resistance of amadoriase and composition for measuring HbA1c using the same |
US11198852B2 (en) | 2014-11-07 | 2021-12-14 | Kikkoman Corporation | Amadoriase having enhanced anionic surfactant tolerance |
JP2021535380A (ja) * | 2018-08-22 | 2021-12-16 | クエスト ダイアグノスティクス インベストメンツ エルエルシー | 糖尿病におけるマイクロサンプリング検出 |
Also Published As
Publication number | Publication date |
---|---|
KR20130107268A (ko) | 2013-10-01 |
CN103124793A (zh) | 2013-05-29 |
BR112013001398A2 (pt) | 2016-05-24 |
JP5871800B2 (ja) | 2016-03-01 |
JPWO2012020744A1 (ja) | 2013-10-28 |
EP2604698A1 (en) | 2013-06-19 |
CA2806261A1 (en) | 2012-02-16 |
US20130171676A1 (en) | 2013-07-04 |
CN103124793B (zh) | 2014-12-31 |
EP2604698A4 (en) | 2014-02-19 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP5871800B2 (ja) | 糖化ヘモグロビンの測定方法 | |
JP6236318B2 (ja) | 糖化ヘモグロビンの測定方法、測定試薬、及び、測定キット | |
JP5955220B2 (ja) | 糖化ヘモグロビンの測定方法 | |
JP7020413B2 (ja) | 糖化ヘモグロビンの測定方法 | |
JP6144208B2 (ja) | アスコルビン酸の影響抑制方法 | |
KR20140114267A (ko) | 측정 대상 성분의 측정 방법 | |
JP7195847B2 (ja) | 糖化蛋白質の測定 | |
WO2021192465A1 (ja) | 異常ヘモグロビン含有試料中の糖化ヘモグロビンの測定方法 | |
WO2021192466A1 (ja) | ヘモグロビンの影響軽減方法、糖化ヘモグロビンの測定方法、糖化ヘモグロビン測定試薬、及び糖化ヘモグロビン測定キット | |
EP3636767A1 (en) | Method for measuring glycated hemoglobin |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
WWE | Wipo information: entry into national phase |
Ref document number: 201180038629.X Country of ref document: CN |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 11816408 Country of ref document: EP Kind code of ref document: A1 |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2012528677 Country of ref document: JP |
|
ENP | Entry into the national phase |
Ref document number: 2806261 Country of ref document: CA |
|
WWE | Wipo information: entry into national phase |
Ref document number: 13811911 Country of ref document: US |
|
ENP | Entry into the national phase |
Ref document number: 20137002549 Country of ref document: KR Kind code of ref document: A |
|
REEP | Request for entry into the european phase |
Ref document number: 2011816408 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2011816408 Country of ref document: EP |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
REG | Reference to national code |
Ref country code: BR Ref legal event code: B01A Ref document number: 112013001398 Country of ref document: BR |
|
ENP | Entry into the national phase |
Ref document number: 112013001398 Country of ref document: BR Kind code of ref document: A2 Effective date: 20130118 |