WO2019216405A1 - Méthode de suppression de la réduction de la sensibilité d'un kit de réactif de mesure de composant biologique - Google Patents

Méthode de suppression de la réduction de la sensibilité d'un kit de réactif de mesure de composant biologique Download PDF

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WO2019216405A1
WO2019216405A1 PCT/JP2019/018704 JP2019018704W WO2019216405A1 WO 2019216405 A1 WO2019216405 A1 WO 2019216405A1 JP 2019018704 W JP2019018704 W JP 2019018704W WO 2019216405 A1 WO2019216405 A1 WO 2019216405A1
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reagent
biological component
measurement
copper
sensitivity
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Japanese (ja)
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研吾 西村
木全　伸介
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東洋紡株式会社
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    • 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
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour

Definitions

  • the present invention relates to a method for suppressing sensitivity reduction of a biological component measurement reagent kit used in clinical diagnosis. More specifically, hydrogen peroxide generated from a biological component using an oxidation-reduction reaction and an oxidation-reduction coloring reagent (such as a Trinder reagent combining an aminoantipyrine compound as a coupler and a hydrogen donor),
  • the present invention relates to a method for suppressing a decrease in sensitivity in a biological component measurement reagent kit that performs colorimetric determination of coloration generated by oxidative condensation in the presence of peroxidase.
  • Examples of the redox coloring reagent system include a method using a hydrogen donor and a coupler.
  • a typical example is a Trinder method in which a hydrogen donor and a coupler are oxidized and condensed with hydrogen peroxide in the presence of peroxidase to form a dye.
  • 4-aminoantipyrine hereinafter also referred to as 4AA
  • 4AA 4-aminoantipyrine
  • the present inventors have prepared an enzyme-peroxidase-coloring agent system biocomponent measurement reagent using an aminoantipyrine compound and a hydrogen donor, and when using it for biocomponent measurement, the measurement sensitivity of unknown cause is reduced. Experienced.
  • an object of the present invention is to provide a means for suppressing a decrease in sensitivity due to 4-hydroxyantipyrine in order to solve the above-mentioned problems that have not been known so far.
  • the present inventors unexpectedly used a copper-containing substance such as a component that generates copper ions in preparing a biological component measurement reagent kit.
  • a copper-containing substance such as a component that generates copper ions
  • the inventors have found that the decrease in sensitivity due to 4-hydroxyantipyrine can be suppressed, and have completed the present invention. That is, the present invention has the following configuration.
  • the biological component measurement method uses a reagent or a reagent set that satisfies the following requirements (a) to (d), and is added to the reagent that satisfies the requirement (d): Item 2.
  • A) An oxidase capable of generating hydrogen peroxide is included.
  • C A redox coloring reagent that reacts with hydrogen peroxide in the presence of peroxidase to develop color.
  • D An aminoantipyrine compound is included as a coupler of the redox coloring reagent.
  • CLAIM ITEM 3
  • Item 4 A method for suppressing a decrease in measurement sensitivity applied to a biological component measurement method using an aminoantipyrine compound as a coupler for a redox coloring reagent, wherein the measurement sensitivity of a biological component is characterized by using a copper-containing substance. How to suppress the decrease.
  • B) Contains peroxidase.
  • C A redox coloring reagent that reacts with hydrogen peroxide in the presence of peroxidase to develop color.
  • An aminoantipyrine compound is included as a coupler of the redox coloring reagent.
  • A An oxidase capable of generating hydrogen peroxide is included.
  • B) Contains peroxidase.
  • a method for producing a reagent kit for measuring a biological component that satisfies the following requirements (a) to (e) and suppresses a decrease in sensitivity due to 4-hydroxyantipyrine comprising: (d) and (e) A method for producing a reagent kit for measuring a biological component, which comprises producing a reagent that satisfies the above requirements as one reagent that simultaneously satisfies two requirements.
  • A An oxidase capable of generating hydrogen peroxide is included.
  • B Contains peroxidase.
  • C A redox coloring reagent that reacts with hydrogen peroxide in the presence of peroxidase to develop color.
  • the present invention it is possible to suppress a decrease in sensitivity due to 4-hydroxyantipyrine in biological component measurement by an enzyme method using an oxidase-peroxidase-color former system. Therefore, according to the present invention, it is possible to measure a favorable biological component. In particular, stable measurement is possible in the measurement of biological components that require high sensitivity, for example, the content of the biological components is extremely small.
  • the biological component measurement sensitivity decrease inhibitor of the present invention is a measurement sensitivity decrease inhibitor applied to a biological component measurement method using an aminoantipyrine compound as a coupler of a redox coloring reagent, and is a copper-containing substance (for example, copper A component that generates ions).
  • the biological component measurement method to which the biological component measurement sensitivity decrease inhibitor of the present invention is applied is preferably a biological component measurement method using a reagent or a reagent set that satisfies the following requirements (a) to (d).
  • A An oxidase capable of generating hydrogen peroxide is included.
  • B Contains peroxidase.
  • C A redox coloring reagent that reacts with hydrogen peroxide in the presence of peroxidase to develop color.
  • D An aminoantipyrine compound is included as a coupler of the redox coloring reagent.
  • a copper-containing substance for example, a component that generates copper ions
  • the aminoantipyrine compound contained in the reagent satisfying the requirement (d) may contain a trace amount of 4-hydroxyantipyrine as a by-product during the production thereof. It has been found that 4-hydroxyantipyrine causes a decrease in measurement sensitivity of biological components. Furthermore, the present inventors have found that such a decrease in sensitivity can be suppressed by allowing a copper-containing substance to coexist with 4-hydroxyantipyrine, thereby completing the present invention.
  • the biological component measurement sensitivity lowering inhibitor of the present invention is preferably used by adding a copper-containing substance to a reagent satisfying the requirement (d), and the biological component measurement sensitivity caused by 4-hydroxyantipyrine. The purpose of this is to suppress the decrease.
  • the reagent or reagent set may be prepared as one reagent that satisfies the requirements of (a) to (d), or is composed of 2 to 3 or more by packaging the reagents. It may be a reagent set.
  • the reagent or reagent set may be in a form such as a kit packaged in a single packaging container, or may be a form in which each reagent is prepared separately and used in a set.
  • the method for suppressing a decrease in measurement sensitivity of a biological component of the present invention is a method for suppressing a decrease in measurement sensitivity applied to a biological component measurement method using an aminoantipyrine compound as a redox coloring reagent coupler, A copper-containing material is used.
  • the method for suppressing a decrease in measurement sensitivity of a biological component of the present invention is a method for suppressing a decrease in measurement sensitivity in a biological component measurement method using a reagent or reagent set that satisfies the following requirements (a) to (d): And a copper-containing substance is added to a reagent that satisfies the requirement (d).
  • A An oxidase capable of generating hydrogen peroxide is included.
  • B Contains peroxidase.
  • C A redox coloring reagent that reacts with hydrogen peroxide in the presence of peroxidase to develop color.
  • D An aminoantipyrine compound is included as a coupler of the redox coloring reagent.
  • the present inventors speculate that the decrease in measurement sensitivity occurs as a result of consumption of hydrogen peroxide by the reaction of 4-hydroxyantipyrine with peroxidase.
  • the reaction of 4-hydroxyantipyrine The present invention was completed by finding that the coexistence can be suppressed by the coexistence of a copper-containing substance (preferably, copper ions that can be generated from the copper-containing substance). That is, the method for reducing the measurement sensitivity of the biological component of the present invention adds the copper-containing substance (for example, a component that generates copper ions) to the reagent that satisfies the requirement (d).
  • a biological component caused by 4-hydroxyantipyrine by coexisting 4-hydroxyantipyrine and a copper-containing substance mixed in a reagent satisfying the above requirements and reacting both to suppress the reactivity of 4-hydroxyantipyrine It is characterized by suppressing a decrease in measurement sensitivity.
  • the time for reacting the copper-containing substance is not particularly limited as long as the effect of the present invention is exhibited. .
  • the time when the copper-containing substance is added to the reagent satisfying the requirement (d) at least at 1 ° C. to 10 ° C. in the state where the copper-containing substance coexists in the reagent satisfying the requirement (d).
  • the storage temperature is preferably a temperature at which the reagent does not freeze, and the upper limit temperature is preferably a temperature at which various components in the reagent do not cause quality deterioration such as alteration or denaturation.
  • the reagent satisfying the requirement (d) is a product reagent (for example, a reagent containing a protein such as an enzyme)
  • a copper-containing substance is added to the product reagent.
  • the reaction proceeds by refrigeration or storage near room temperature in the coexistence of a copper-containing substance, and storage before use, so that the reagent that satisfies the requirement (d) before use What is necessary is just to use it, after at least 2 weeks have passed since the date of manufacture of the biological component measurement reagent kit manufactured by adding a copper-containing substance.
  • a reagent that satisfies the requirement (d) is an intermediate reagent (for example, an aminoantipyrine compound having a concentration several to several tens of times that of a product reagent as a buffer component).
  • an adjusted reagent for example, after adding a copper-containing substance to the intermediate reagent, for example, store it for several days at a temperature condition of 30 ° C. or more, and for several hours at a temperature condition of 45 ° C. or more.
  • a product reagent may be prepared.
  • a copper-containing substance coexisting in the reagent that satisfies the requirement of (d) is added. Except for the product reagent.
  • the present inventors have found that such a decrease in measurement sensitivity caused by 4-hydroxyantipyrine can be suppressed in advance by coexisting a copper-containing substance with 4-hydroxyantipyrine. Although the mechanism by which the copper-containing substance suppresses the sensitivity decrease due to 4-hydroxyantipyrine is not clear, the present inventors have caused some structural change of 4-hydroxyantipyrine due to the copper-containing substance (for example, copper ions that can be generated in the future). It is assumed that hydrogen peroxide is not consumed without reacting with peroxidase.
  • the structural change of 4-hydroxyantipyrine is presumed to be an irreversible change, and the decrease in the measurement sensitivity of biological components caused by 4-hydroxyantipyrine suppressed by the copper-containing substance is The present inventors presume that the reaction between 4-hydroxyantipyrine and peroxidase will not be restored even if the copper-containing material (copper ions that can be generated in the specific embodiment) is removed from the medium.
  • the concentration of the copper-containing substance coexisting in the reagent satisfying the requirement (d) is preferably 0.00001 to 1 mM, and 0.00003 to 0.00. 3 mM is more preferable, 0.0001 to 0.1 mM is more preferable, and 0.0001 to 0.01 mM is particularly preferable.
  • concentration of the copper-containing substance is less than 0.00001 mM, the inhibitory effect is small, and when the concentration of the copper-containing substance is more than 1 mM, the color caused by the copper-containing substance increases in the biological component measurement reaction, resulting in a measurement error.
  • the preferable concentration range of the copper-containing substance coexisting in the reagent satisfying the requirement (d) depends on the concentration of 4-hydroxyantipyrine mixed in the reagent satisfying the requirement (d). -If the concentration of hydroxyantipyrine is low, a decrease in the measurement sensitivity of biological components can be suppressed even if the concentration of copper-containing material is low. If the concentration of 4-hydroxyantipyrine is high, it is effective to use a higher concentration of copper-containing material. Therefore, in the biological component measurement sensitivity reduction suppressing method of the present invention, the concentration of the copper-containing substance is preferably set as appropriate according to the concentration of 4-hydroxyantipyrine mixed therein.
  • ICP emission spectroscopy ICP-AES
  • ICP mass spectrometry ICP-MS
  • concentration of copper ions can be measured by ICP-AES (using SPECTROBLUE manufactured by Ametech).
  • ICP-MS using Agilent 7700s ICP-MS manufactured by Agilent Technologies.
  • the concentration of 4-hydroxyantipyrine present in the reagent that satisfies the requirement (d) is added with a copper-containing substance (for example, a component that generates copper ions).
  • a copper-containing substance for example, a component that generates copper ions.
  • the concentration is about 0.1 to 50 ⁇ g / ml, the effect is particularly easily obtained.
  • the concentration of 4-hydroxyantipyrine present in the reagent that satisfies the requirement (d) before adding the copper-containing substance is less than 0.1 ⁇ g / ml, the measurement sensitivity of biological components is reduced by 4-hydroxyantipyrine Is 1% or less, and there is little need to suppress a decrease in measurement sensitivity by coexisting a copper-containing substance.
  • the concentration of 4-hydroxyantipyrine present in the reagent that satisfies the requirement (d) before adding the copper-containing substance is more than 50 ⁇ g / ml, the decrease in measurement sensitivity of biological components due to 4-hydroxyantipyrine is suppressed.
  • a high concentration of copper-containing material is required, and therefore, color development tends to increase due to side reactions caused by the copper-containing material, which tends to cause measurement errors.
  • a general aminoantipyrine compound (4-aminoantipyrine) drug substance contains about 0.005 to 0.30 w / w% of 4-hydroxyantipyrine. is doing.
  • the present invention is a reagent containing about 0.01 to 100 g / l of aminoantipyrine compound (4-aminoantipyrine), preferably about 0.01 to 10 g / l of aminoantipyrine compound (4-amino It is effective to be used for a reagent containing an antipyrine), more preferably a reagent containing about 0.01 to 5 g / l of an aminoantipyrine compound (4-aminoantipyrine).
  • the amount of 4-hydroxyantipyrine mixed in it is preferable to remove 4-hydroxyantipyrine from the aminoantipyrine compound in advance by a method for removing 4-hydroxyantipyrine, which will be described later, and then subject it to biological component measurement. .
  • the biological component to be measured by the biological component measurement reagent kit of the present invention is not particularly limited, and can be used for measurement of various biological components.
  • the biological components used in the biological component measurement of the present invention are uric acid (UA), creatinine (CRE), triglyceride (TG), cholesterol (CHO), AST (GOT), ALT (GPT), LDH (lactic acid dehydrogenation).
  • Enzymes and isozymes ALP (alkaline phosphatase) and isozymes, CK (creatine kinase) and isozymes, amylases (Amy) and isozymes, lipases, ⁇ -GTP ( ⁇ -glutamyl transpeptidase), cholinesterase (ChE), sodium (Na) , Potassium (K), chlor (Cl), calcium (Ca), phosphorus (P) [inorganic phosphorus (IP)], iron (Fe), magnesium (Mg), total protein (TP), serum protein fraction (PF) ), Urea nitrogen (BUN), creatinine (CRE), uric acid (U ), Bilirubin (Bil), ammonia, cholesterol, HDL cholesterol (HDL-C, high density lipoprotein cholesterol), LDL cholesterol (LDL-C, low density lipoprotein cholesterol), neutral fat (triglyceride) (TG), cholesterol (CHO), BTR (BTR, total branched chain amino acid / t
  • uric acid U
  • CRE creatinine
  • TG triglyceride
  • HbA1c glycated hemoglobin
  • uric acid In the case of measuring uric acid (UA), hydrogen peroxide produced by the reaction of uricase (oxidase) using uric acid (UA) as a substrate can be quantified by a peroxidase-chromogenic system.
  • creatinine (CRE) When measuring creatinine (CRE), hydrogen peroxide is not directly generated in the reaction of creatinine amidinohydrolase using creatinine (CRE) as a substrate. Therefore, creatine obtained by adding creatine generated in the reaction of creatinine amidinohydrolase to the reagent in advance is used.
  • Peroxidase-coloring agent system by designing a so-called conjugation reaction in which sarcosine is generated by reacting with amide hydrolase, and hydrogen peroxide is generated using sarcosine oxidase (oxidase) with sarcosine added to the reagent in advance. Quantification of creatinine (CRE) concentration by means of
  • triglyceride (TG) When triglyceride (TG) is measured, hydrogen peroxide is generated by using lipoprotein lipase using triglyceride (TG) as a substrate and glycerol kinase and glycerol triphosphate oxidase (oxidase) as conjugate enzymes.
  • TG triglyceride
  • the triglyceride (TG) concentration can be quantified using a peroxidase-color former system.
  • HbA1c glycated hemoglobin
  • glycated hemoglobin oxidase for example, fructosyl amino acid oxidase
  • creatinine (CRE) and glycated hemoglobin (HbA1c) have extremely low biological component content, and therefore, particularly sensitive measurement is required.
  • the influence of 4-hydroxyantipyrine which inhibits the reaction of the oxidase-peroxidase-color former system, can be suppressed and the decrease in sensitivity of biological component measurement can be suppressed, and thus high sensitivity measurement is required.
  • the present invention is suitable for measurement of creatinine and glycated hemoglobin, and in particular, it is suitably used for measurement of creatinine.
  • the biological component measurement kit of the present invention includes a measurement sensitivity lowering inhibitor containing a copper-containing substance as described above.
  • the biological component measurement kit of the present invention satisfies the following requirements (a) to (e) and simultaneously satisfies the two requirements for a reagent that satisfies the following requirements (d) and (e): It is characterized by being a single reagent.
  • A An oxidase capable of generating hydrogen peroxide is included.
  • B Contains peroxidase.
  • C A redox coloring reagent that reacts with hydrogen peroxide in the presence of peroxidase to develop color.
  • D An aminoantipyrine compound is included as a coupler for the redox coloring reagent.
  • E Contains a copper-containing material.
  • the biological component measurement kit of the present invention it is preferable that one reagent that satisfies the two requirements (d) and (e) has passed at least one month after production.
  • the present inventors presume that 4-hydroxyantipyrine reacts with peroxidase, and as a result, hydrogen peroxide is consumed.
  • the reactivity of 4-hydroxyantipyrine is determined by adding a copper-containing substance to 4-hydroxyantipyrine. It discovered in this invention that it can suppress by adding.
  • the reaction rate of the suppression reaction may affect the reaction temperature and reaction time.
  • the suppression effect is produced when the storage period is several weeks to one month or more. It is preferable that at least one month has passed since the biological component measurement kit of the present invention. In general, such a measurement kit is considered to take one month to several months after being manufactured and used through an inventory / distribution process, during which the suppression reaction proceeds and the effect of the present invention is exhibited. It will be.
  • the biological component measuring method of the present invention is characterized by measuring biological components using the aforementioned biological component measuring kit.
  • the method for producing a biological component measurement kit of the present invention is a method for producing a biological component measurement reagent kit that satisfies the following requirements (a) to (e) and suppresses a decrease in sensitivity due to 4-hydroxyantipyrine. , (D) and (e) are manufactured as one reagent that satisfies the two requirements at the same time.
  • B) Contains peroxidase.
  • C A redox coloring reagent that reacts with hydrogen peroxide in the presence of peroxidase to develop color.
  • (D) An aminoantipyrine compound is included as a coupler of the redox coloring reagent.
  • E) Contains a copper-containing material.
  • the reagent that satisfies the requirements (d) and (e) is preferably manufactured as one reagent that simultaneously satisfies the two requirements, and that the biological component derived from 4-hydroxyantipyrine is measured.
  • a sufficient period for example, about one month or more
  • one reagent that simultaneously satisfies the two requirements is an intermediate reagent in the production process of the biological component measurement reagent kit.
  • the product reagent may be prepared using the intermediate reagent.
  • the biological component measurement kit of the present invention needs to suppress the reactivity of 4-hydroxyantipyrine by adding a copper-containing substance to 4-hydroxyantipyrine. ) May be carried out by allowing a copper-containing substance to coexist with 4-hydroxyantipyrine mixed in a reagent that satisfies the requirements of (1), but this suppression reaction is not necessarily carried out in the product reagent. For example, an aminoantipyrine compound is added.
  • a product reagent may be produced using the intermediate reagent and provided as a biological component measurement kit.
  • the reagent intermediate containing an aminoantipyrine compound as described above is, for example, diluting the aminoantipyrine compound base with a buffer solution or the like in the step of producing a reagent containing the aminoantipyrine compound. It means an intermediate reagent adjusted to a concentration several times to several tens of times that of the product reagent.
  • the biological component measurement kit of the present invention when adding a copper-containing substance to an intermediate reagent containing an aminoantipyrine compound and performing an inhibitory reaction, adds the copper-containing substance to the intermediate reagent and then ends the product production date.
  • the sum of the period and the period from product manufacture date to product use date is preferably at least one month.
  • the copper-containing substance of the present invention (for example, a component that generates copper ions) is allowed to coexist with 4-hydroxyantipyrine mixed as an impurity of an aminoantipyrine compound, thereby measuring the biological component attributed to 4-hydroxyantipyrine. Presumed to suppress the decrease. Although it is not clear why the coexistence of the copper-containing substance of the present invention with 4-hydroxyantipyrine suppresses a decrease in the sensitivity of biological component measurement, there are 4 copper-containing substances (for example, copper ions that can be generated from this).
  • the copper-containing substance used in the present invention is preferably a component that generates copper ions, and is not particularly limited. Examples thereof include copper salts containing copper ions, copper proteins containing copper ions as prosthetic factors, and pigments containing copper ions. Can do.
  • These components that generate copper ions can be appropriately selected according to, for example, a biological component or a biological sample to be measured. From the viewpoint of excellent handling convenience and the expectation that even higher effects can be exhibited, copper chloride (I), copper chloride (II), copper acetate, copper sulfate, copper nitrate, proclin 200, ascorbic acid oxidation It is preferable to use an enzyme.
  • the usage-amount of the copper containing material used for this invention is not specifically limited as long as there exists an effect of this invention.
  • the copper-containing substance is preferably used as a product reagent mixed with an aminoantipyrine compound intermediate reagent or an aminoantipyrine compound, and as an example, the copper reagent in the intermediate reagent or product reagent
  • the concentration of the contained substance may be adjusted so as to be 0.00001 to 1 mM, and the concentration of the copper-containing substance in the reagent intermediate or reagent is preferably 0.00003 to 0.3 mM, more preferably 0.8.
  • the blending amount may be adjusted to 0001 to 0.1 mM, more preferably 0.0001 to 0.01 mM.
  • the copper-containing substance used in the present invention has an effect of suppressing the decrease in the sensitivity of biological component measurement caused by 4-hydroxyantipyrine contained as an impurity of the aminoantipyrine compound after being blended with the aminoantipyrine compound. Is preferably allowed to elapse for a certain time after blending.
  • the storage temperature is 1 ° C. to 10 ° C. for 2 weeks or more, 11 ° C.
  • the longer the time is required depending on the storage temperature, the higher the temperature, the shorter and the lower the temperature.
  • the upper limit of time to make it react is not specifically limited, For example, it can be made into 10 years or less.
  • the copper-containing substance used in the present invention suppresses a decrease in sensitivity of biological component measurement due to 4-hydroxyantipyrine as long as the quality of the reagent used in the biological component measurement kit does not deteriorate. The effect is maintained.
  • the product life cycle of a general biological component measurement kit it takes about several weeks or more than one month until it is blended with reagents in the production process and used for quality testing, shipping, distribution, storage and use.
  • the measurement kit including the reagent in which the copper-containing substance of the present invention is coexistent with the aminoantipyrine compound is in a state where the sensitivity reduction suppressing effect by the copper-containing substance used in the present invention is expressed and maintained at the stage of use.
  • the product life cycle when the product life cycle is shortened, it can be prepared by manufacturing process management, product management at the time of shipment, or the like.
  • oxidase used in the present invention can generate hydrogen peroxide from a substrate, it can be used without limitation depending on the target measurement target. Specific examples include, but are not limited to, uricase, sarcosine oxidase, glycerol triphosphate oxidase, fructosyl amino acid oxidase and the like. As commercially available products, UAO-211 (manufactured by Toyobo), SAO-351 (manufactured by Toyobo), G3O-311 (manufactured by Toyobo) and the like are preferably used. There are no particular restrictions on the amount used or the form of addition.
  • peroxidase As the peroxidase used in the present invention, any kind of enzyme may be used as long as it catalyzes the reaction between hydrogen peroxide and a redox coloring reagent.
  • peroxidases derived from plants, bacteria, and basidiomycetes Is mentioned.
  • horseradish, rice, soybean-derived peroxidase is preferable, and horseradish-derived peroxidase is more preferable because of purity, availability, and price.
  • PEO-131 (Toyobo), PEO-301 (Toyobo), PEO-302 (Toyobo) and the like are preferably used as commercially available products. There are no particular restrictions on the amount used or the form of addition.
  • Peroxidase activity is defined by the following method. 14 mL of distilled water, 2 mL of 5% (W / V) pyrogallol aqueous solution, 1 mL of 0.147M hydrogen peroxide solution and 2 mL of 100 mM phosphate buffer solution (pH 6.0) were sequentially mixed, and then pre-conditioned at 20 ° C. for 5 minutes. Then, 1 mL of the sample solution is added to start the enzyme reaction. After reacting for 20 seconds, the reaction is stopped by adding 1 mL of 2N aqueous sulfuric acid, and the resulting purpurogallin is extracted 5 times with 15 mL of ether.
  • the total volume is 100 mL, and the absorbance at a wavelength of 420 nm is measured ( ⁇ OD test ).
  • the absorbance at a wavelength of 420 nm is measured ( ⁇ OD test ).
  • 14 mL of distilled water, 2 mL of 5% pyrogallol solution, 1 mL of 0.147M hydrogen peroxide solution and 2 mL of 100 mM phosphate buffer (pH 6.0) were mixed in order, and then 1 mL of 2N sulfuric acid aqueous solution was added and mixed. Then, add 1 mL of the sample solution. About this liquid, ether extraction is performed in the same manner as described above, and the absorbance is measured ( ⁇ OD blank ).
  • the amount of purpurogallin produced is calculated from the difference in absorbance between ⁇ OD test and ⁇ OD blank , and the peroxidase activity is calculated.
  • the amount of enzyme that produces 1.0 mg of purpurogallin in 20 seconds under the above conditions is defined as 1 purpurogallin unit (U).
  • the calculation formula is as follows.
  • Peroxidase activity (U / mg) peroxidase activity (U / mL) ⁇ 1 / C 0.117: Absorbance at 420 nm of 1 mg% purpurogallin ether solution
  • C Enzyme concentration at dissolution (c mg / mL) (One propargalin unit corresponds to 13.5 international units (with o-dianisidine as a substrate and reaction conditions at 25 ° C.).
  • the sample solution was dissolved in 0.1 M phosphate buffer pH 6.0 that had been ice-cooled in advance, and diluted to 3.0 to 6.0 purpurogallin units (U) / mL with the same buffer solution. Then, it is preferable to use for the measurement.
  • Redox coloring reagent As the redox coloring reagent used in the measurement of the biological component of the present invention, any kind of dye may be used as long as it reacts with hydrogen peroxide and develops color. For example, a combination of a hydrogen donor and a coupler is used. Can be mentioned. There are no particular restrictions on the amount used or the form of addition. All of these can be obtained as commercial products.
  • a typical example using a hydrogen donor and a coupler is a Trinder method in which a hydrogen donor and a coupler are oxidized and condensed with hydrogen peroxide in the presence of peroxidase to form a dye.
  • phenol, a phenol derivative, an aniline derivative, naphthol, a naphthol derivative, a naphthylamine, a naphthylamine derivative, or the like is used as a hydrogen donor used in the Trinder method or the like.
  • Coupler These hydrogen donors can be used in combination with a coupler.
  • couplers examples include 4-aminoantipyrine (4AA), aminoantipyrine compounds such as aminoantipyrine derivatives; vanillin diamine sulfonic acid compounds such as vanillin diamine sulfonic acid; methylbenzthiazolinone hydrazone (MBTH), sulfonated methylbenzthia.
  • MBTH methylbenzthiazolinone hydrazone
  • SMBTH zolinone hydrazone
  • the present invention can suppress a decrease in sensitivity of a biological component measurement reagent kit caused by a trace amount of 4-hydroxyantipyrine contained in an aminoantipyrine compound, and is effective when an aminoantipyrine compound is used as a coupler. is there.
  • the present invention is useful when 4-aminoantipyrine is used as a coupler.
  • the coupler used in the present invention may be two or more aminoantipyrine compounds, or may be used in combination with another coupler in addition to the aminoantipyrine compound, but preferably one aminoantipyrine compound. It is preferable to use 4-aminoantipyrine.
  • the amount of aminoantipyrine compound used in the present invention is not particularly limited as long as the effects of the present invention are exhibited.
  • the aminoantipyrine compound may contain a trace amount of 4-hydroxyantipyrine that causes a decrease in sensitivity, and it is desirable to reduce the amount of 4-hydroxyantipyrine. By coexisting, the sensitivity reduction can be suppressed even if some amount of 4-hydroxyantipyrine is contained.
  • the concentration of 4-hydroxyantipyrine mixed in the reagent containing the aminoantipyrine compound before coexisting with the copper-containing substance is preferably 0.1 to 50 ⁇ g / ml, more preferably It is effective to prepare and use the aminoantipyrine compound used for the biological component measurement so that the amount is 0.3 to 20 ⁇ g / ml, particularly preferably 1 to 10 ⁇ g / ml.
  • concentration of 4-hydroxyantipyrine is lower than the above range, there is little influence of a decrease in measurement sensitivity of biological components due to 4-hydroxyantipyrine, and there is little need to suppress a decrease in measurement sensitivity due to the coexistence of a copper-containing substance.
  • the concentration of 4-hydroxyantipyrine exceeds the above range, a high concentration of copper-containing material is required to suppress a decrease in measurement sensitivity of biological components due to 4-hydroxyantipyrine. This tends to reduce the measurement accuracy of biological components.
  • the amount of 4-hydroxyantipyrine mixed in is large, remove 4-hydroxyantipyrine mixed in the aminoantipyrine compound in advance by a method for removing 4-hydroxyantipyrine, which will be described later. Is preferred.
  • the present invention has an aminoantipyrine compound (4-aminoantipyrine) of about 0.01 to 100 g / l.
  • a reagent containing preferably about 0.01 to 10 g / l of an aminoantipyrine compound (4-aminoantipyrine), more preferably about 0.01 to 5 g / l of an aminoantipyrine compound (4-aminoantipyrine) It is effective to use for a reagent containing. By using an aminoantipyrine compound in such a range, the effects of the present invention can be more effectively exhibited.
  • the content of 4-hydroxyantipyrine in the aminoantipyrine compound is, for example, high performance liquid chromatography (hereinafter also referred to as HPLC method), gas chromatography (hereinafter also referred to as GC method), mass spectrometry (hereinafter referred to as MS). It can also be measured by performing quantitative methods such as a nuclear magnetic resonance method (hereinafter also referred to as NMR method) alone or in any combination.
  • HPLC method is preferably used from the viewpoint of the convenience of operation and the economical efficiency of the system / equipment.
  • the HPLC method column is preferably a reverse phase column, and more preferably a silica-based porous column.
  • (4-hydroxyantipyrine) 4-hydroxyantipyrine has a structure in which the amino group at the 4-position of 4-aminoantipyrine is converted to a hydroxyl group, and is considered to be a byproduct produced and mixed in the production process of 4-aminoantipyrine.
  • 4-hydroxyantipyrine has a great influence on the color reaction in the color development reaction in the biological component measurement method even when the amount of contamination is very small. This phenomenon is caused by the fact that 4-hydroxyantipyrine has a faster reaction rate and consumes hydrogen peroxide than aminoantipyrine compounds originally intended to be reacted as a coupler (for example, 4-aminoantipyrine). It is guessed.
  • an aminoantipyrine compound drug substance that can contain 4-hydroxyantipyrine may be used as it is, or an aminoantipyrine compound drug substance that has a low 4-hydroxyantipyrine content. May be used by selecting the amount of 4-hydroxyantipyrine content reduced by removing 4-hydroxyantipyrine from the aminoantipyrine compound drug substance. .
  • the method for reducing the 4-hydroxyantipyrine content from an aminoantipyrine compound there is no particular limitation on the method for reducing the 4-hydroxyantipyrine content from an aminoantipyrine compound.
  • any method known in the art such as a method of using chromatography such as HPLC from a biological component measurement reagent, a method of dissolving 4-hydroxyantipyrine on an adsorbent such as a resin after being dissolved in water or a solvent, etc.
  • Separation / removal may be performed using means.
  • chromatography is used as a means for removing 4-hydroxyantipyrine from an aminoantipyrine compound
  • the physicochemical principle of the separation is not particularly limited. Examples include various principles such as distribution (normal phase / reverse phase), adsorption, molecular exclusion, and ion exchange. Separation may be performed by a method in which a ligand is bound to a resin or an adsorbent.
  • the 4-hydroxyantipyrine removal means general reverse phase chromatography can be used.
  • the carrier for reverse phase chromatography is not particularly limited.
  • silica gel is suitable, but a polymer carrier may be used. When silica gel is used as a carrier, those with and without end cap treatment can be selected.
  • the chromatography apparatus can be used by appropriately preparing the conditions according to the purpose regardless of whether the chromatography system is low pressure, medium pressure, or high pressure.
  • the type of ligand that binds to the chromatography carrier is not particularly limited. In addition to the octadecyl group (ODS), which is widely used, a ligand and an octyl group can be selected by optimizing the conditions.
  • ODS octadecyl group
  • Ligand binding may be monomeric or polymeric. Any filler can be used by optimizing the separation conditions.
  • the chromatographic mobile phase may be water and a water-soluble solvent such as methanol or acetonitrile.
  • the pH of the chromatographic mobile phase should be acidic.
  • a small amount of preparation or ion pair reagent may be added to the side.
  • the mobile phase flow rate may be optimized according to the capacity of the system used. Further, elution may be performed stepwise instead of using a linear gradient.
  • the biological component measurement reagent kit of the present invention preferably contains a reagent containing a buffer component.
  • ascorbate oxidase, preservatives, salts, enzyme stabilizers, chromogen stabilizers, etc. are added to the reagents contained in the biological component measurement reagent kit of the present invention as long as they do not affect the reaction. Also good.
  • buffer solution component examples include Tris buffer solution, phosphate buffer solution, borate buffer solution, carbonate buffer solution, and GOOD buffer solution. There are no particular limitations on the amount used, the set pH, the form of addition, and the like. All of these can be obtained as commercial products.
  • GOOD buffers include N- (2-acetamido) -2-aminoethanesulfonic acid (ACES), N, N-bis (2-hydroxyethyl) -2-aminoethanesulfonic acid (BES), N-cyclohexyl- 2-aminoethanesulfonic acid (CHES), 2- [4- (2-hydroxyethyl) -1-piperazinyl] ethanesulfonic acid (HEPES), 2-morpholinoethanesulfonic acid (MES), piperazine-1,4-bis (2-ethanesulfonic acid) (PIPES), N-tris (hydroxymethyl) methyl-2-aminomethanesulfonic acid (TES), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), N-cyclohexyl-2- Hydroxy-3-aminopropanesulfonic acid (CAPSO), 3- [N, N-bis (2-hydro Cyethy
  • biological component measurement reagent of the present invention there are no particular limitations on the amount used or the form of addition of ascorbic acid oxidase, preservatives, salts, enzyme stabilizers, chromogen stabilizers, and the like. All of these can be obtained as commercial products.
  • preservatives include Procrine 150, Procrine 200, Procrine 300, Procrine 950, azide, chelating agent, antibiotic, antibacterial agent and the like.
  • Examples of the chelating agent include ethylenediaminetetraacetic acid and its salt.
  • Antibiotics include gentamicin, kanamycin, chloramphenicol and the like.
  • antibacterial agents examples include methyl isothiazolinone and imidazolidinyl urea.
  • salts examples include sodium chloride, potassium chloride, aluminum chloride and the like.
  • Enzyme stabilizers include sucrose, trehalose, cyclodextrin, gluconate, amino acids and the like.
  • chromogen stabilizers examples include chelating agents such as ethylenediaminetetraacetic acid and its salts, and cyclodextrins.
  • the reagent contained in the biological component measurement reagent kit of the present invention may be a liquid reagent dissolved in an arbitrary solvent (for example, purified water, organic solvent, etc.), or dissolved in the same solvent as described above before use. It may be a dry powder reagent (for example, freeze-dried powder) used.
  • a general-purpose automatic analyzer for example, Hitachi 7180 automatic analyzer
  • the biological component measurement kit of the present invention may be configured to be applicable to such an automatic analyzer.
  • the mode is not particularly limited.
  • Various forms such as a kit called a so-called dry system and a kit using a sensor can be exemplified.
  • kits composed of one reagent or a kit composed of 2 to 3 or more by packaging the reagent is used.
  • a reagent is packaged to form a kit composed of two or more reagents, for example, (a) an oxidase capable of generating hydrogen peroxide and (c) a hydrogen donor of the redox coloring reagent.
  • the kit may be configured by being packaged into a reagent containing 4-aminoantipyrine as a coupler among (b) peroxidase and (d) redox coloring dye.
  • kits composed of a liquid reagent in which two reagents are packaged (hereinafter also referred to as a two-reagent liquid reagent) will be described.
  • the first type of reagent (hereinafter also referred to as the first reagent or R1) is added to the sample and allowed to react for a certain period of time, and then the second type.
  • the target component can be quantified by further adding and reacting the reagent (hereinafter also referred to as the second reagent or R2) and measuring the change in absorbance during this period.
  • the biological component measurement reagent of the present invention is supplied in two or more packages in consideration of application to an automatic analyzer as described above, for example, the concentration of the copper-containing substance in each packaging reagent or It is determined whether or not the concentration of the other components is within the preferable concentration range of each component.
  • the biological component measurement method targeted by the present invention includes the following steps (1) to (3).
  • (1) a step of generating hydrogen peroxide by causing an oxidase to act on a biological component; (2)
  • the hydrogen peroxide generated in step (1) reacts with peroxidase in the presence of peroxidase, whereby the reaction solution is colored by oxidative condensation of 4-aminoantipyrine and redox coloring reagent.
  • Process (3) A step of colorimetrically determining the reaction product colored in step (2).
  • the biological component measurement method of the present invention is a biological component measurement method by an enzyme method, particularly a method using an oxidase-peroxidase-color former system, that is, the amount of biological component by enzymatic reaction of the biological component in the specimen.
  • the measurement principle is to perform colorimetric determination of the color produced by generating hydrogen peroxide according to the above and reacting it with a color former in the presence of peroxidase.
  • Biological component measurement methods using this principle have already been established in the art. Therefore, the knowledge can be applied to the present invention to measure the amount or concentration of a biological component in various samples, and the mode is not particularly limited.
  • the present inventors have found that 4-hydroxyantipyrine contained in a trace amount in an active ingredient of an aminoantipyrine compound (especially 4-aminoantipyrine) inhibits the reaction of this oxidase-peroxidase-color-developer system. We found for the first time that it caused a decline.
  • the mechanism by which 4-hydroxyantipyrine inhibits the above reaction is not necessarily clear, but its structure causes a redox coloring reagent and 4-hydroxyantipyrine to undergo a condensation reaction in the presence of hydrogen peroxide by the action of peroxidase. It is guessed that it consumes.
  • specimen containing a biological component used for the biological component measurement of the present invention examples include biological fluids such as blood (particularly serum and plasma), urine, ascites, and cerebrospinal fluid, and people such as beverages and foods. Ingestion and so on.
  • biological fluids such as blood (particularly serum and plasma), urine, ascites, and cerebrospinal fluid, and people such as beverages and foods. Ingestion and so on.
  • human body fluid a sample derived from blood such as serum or plasma, a sample derived from urine, or the like
  • the present invention uses a copper-containing substance, and in comparison with the case where no copper-containing substance is used, the detection sensitivity due to 4-hydroxyantipyrine in the measurement of biological components by an enzyme method using an oxidase-peroxidase-color former system. Can be suppressed.
  • the detection sensitivity of the biological component measurement reagent kit will be described using creatinine as an example of the biological component.
  • eGFR also referred to as estimated glomerular filtration rate
  • eGFR has been required to have a measurement accuracy of creatinine up to two digits after the decimal point, and a minimum detection sensitivity of about 0.03 mg / dL is required as a creatinine concentration.
  • the present invention by using a copper-containing substance, it is possible to suppress a decrease in sensitivity of the biological component measurement reagent kit caused by 4-hydroxyantipyrine and increase detection sensitivity to the above level.
  • Example 1 Separation of 4HA by HPLC Since it was found that the degree of decrease in reagent sensitivity was due to the difference in lots of 4AA, it was assumed that the content of impurities contained in trace amounts in 4AA differed from lot to lot, and impurities were determined by the aforementioned 4HA quantitative method (HPLC method). Was detected. The purity of 4AA used for examination of lot difference was 98.0% or more according to JIS-K8048.
  • the HPLC fraction when 4AA is analyzed is shown in FIG.
  • the large peak that is observed around 7 to 8 minutes on the horizontal axis of the graph is the peak of 4-aminoantipyrine.
  • this HPLC fraction as shown in FIG. 1, three kinds of impurities other than 4-aminoantipyrine were confirmed.
  • Example 2 Identification of 4HA Examining the contents of each lot for the three types of impurities (a, b, c) seen in FIG. 1 of Example 1, it was found that the contents of only the impurities b differ greatly from lot to lot. Therefore, the molecular weight and structure of these three kinds of substances were analyzed by mass spectrometry (MS spectrum method).
  • FIG. 2 shows the structural formula of 4HA.
  • Example 3 Dependence of 4HA concentration on creatinine measurement sensitivity Using creatinine as a biological component, the dependency of 4HA on the amount of contamination was evaluated with respect to a decrease in measurement sensitivity caused by 4HA mixed in the reagent.
  • Each measurement reagent was prepared by adding 4HA to the second reagent of the following creatinine measurement reagent so that the final concentration in the reagent would be 0.13 to 8.75 ⁇ g / ml.
  • a 5 mg / dL creatinine aqueous solution was used as the biological component sample.
  • a creatinine measuring reagent having the following composition was prepared.
  • 4-aminoantipyrine commercially available 4-aminoantipyrine raw material was purified to produce 4-aminoantipyrine free of 4-hydroxyantipyrine and used.
  • Example 4 Sensitivity reduction suppression effect by copper salt About the copper salt, the inhibitory effect of the sensitivity fall resulting from 4HA was confirmed.
  • the experimental conditions were as follows. For the reagents other than the control (a), 4HA was added to the second reagent of the creatinine measurement reagent so that the final concentration in the reagent was 10 ⁇ g / ml, and a 5 mg / dL creatinine aqueous solution was used as the biological component sample. The process was performed under the same conditions as in Example 3 except that.
  • Second reagent stored for 3 days under the respective refrigeration conditions (6 ° C) and accelerated test (35 ° C) temperature conditions after adding various copper salts to the final concentration in the second reagent of the specified creatinine measurement reagent was used to examine measurement sensitivity (mABS).
  • the blank value of each reagent was also measured at the same time, and a value obtained by subtracting the blank value from the measurement sensitivity was calculated as STD sensitivity.
  • the results are shown in Table 2.
  • the decrease in sensitivity due to 4HA can be effectively suppressed by coexisting various copper salts. That is, the measurement sensitivity of the control (b) to which 4HA was added to 10 ⁇ g / ml was 32% for 3 days of refrigeration and 45% for 3 days at 35 ° C. The measurement sensitivity was 75% to 100% after 3 days of refrigeration, and 71% to 100% after 3 days at 35 ° C. From this result, it can be seen that, as a general tendency, storage at 35 ° C. is more stable and a higher sensitivity reduction suppressing effect can be obtained more easily than storage under refrigerated conditions. Since the evaluation at 35 ° C.
  • Example 5 Inhibition of sensitivity reduction by a mixture containing copper protein / copper salt Using Procrine 200 as a mixture containing ASO-311 (ascorbic acid oxidase) as a copper protein and a copper salt, the effect of suppressing the decrease in sensitivity due to 4HA was confirmed.
  • the experimental conditions were as follows. For the reagents other than the control (a), 4HA was added to the second reagent of the creatinine measurement reagent so that the final concentration in the reagent was 10 ⁇ g / ml, and a 5 mg / dL creatinine aqueous solution was used as the biological component sample. The process was performed under the same conditions as in Example 3 except that.
  • Second reagent stored for 3 days under the temperature conditions of the accelerated test (35 ° C) after adding ASO-311 and Procrine 200 to the second reagent of the creatinine measurement reagent so as to have a final concentration in the predetermined second reagent was used to examine measurement sensitivity (mABS).
  • the blank value of each reagent was also measured at the same time, and a value obtained by subtracting the blank value from the measurement sensitivity was calculated as STD sensitivity.
  • the ratio of the measurement sensitivity (mABS) of each reagent to the measurement sensitivity (mABS) of the control (a) to which 4HA was not added [vs control (%)] was calculated. The results are shown in Table 3.
  • ASO-311 (ascorbate oxidase) is known to be a copper protein containing copper ions.
  • the procrine 200 may contain a copper salt as a stabilizer in addition to the main components 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one.
  • a mixture containing copper protein or copper salt is known. Therefore, even when such a mixture containing copper protein or copper salt was used, it was confirmed that the effects of the present invention were exhibited. That is, the measurement sensitivity of the control (b) to which 4HA was added to 10 ⁇ g / ml was 41% at 35 ° C. for 3 days, whereas the measurement sensitivity was 35 ° C. 3 in the result using ASO-311. The measurement sensitivity was 96% to 98% at 35 ° C. for 3 days.
  • Example 6 Concentration dependence of copper salt / 4HA on the effect of suppressing the decrease in sensitivity The copper salt and the concentration dependency of 4HA were confirmed with respect to the effect of suppressing the decrease in sensitivity due to 4HA.
  • the experimental conditions were as follows. For reagents other than the control (a), 4AA was used as the second reagent in the creatinine measurement reagent as the final concentration in the second reagent, and 600 ⁇ g / ml, 300 ⁇ g / ml, 100 ⁇ g / ml, and 4HA in the second reagent.
  • the final conditions were 10 ⁇ g / ml, 5 ⁇ g / ml, and 1.6 ⁇ g / ml, and a 5 mg / dL creatinine aqueous solution was used as a biological component sample.
  • each was stored for 3 days under the temperature condition of the accelerated test (35 ° C.).
  • the measurement sensitivity (mABS) was examined.
  • the blank value of each reagent was also measured at the same time, and a value obtained by subtracting the blank value from the measurement sensitivity was calculated as STD sensitivity.
  • the present invention can be applied to a measurement method for measuring a biological component using an oxidation-reduction reaction, and a reagent or composition used in the method.

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Abstract

La présente invention concerne un agent pour supprimer une réduction de la sensibilité de mesure, l'agent étant appliqué à une méthode de mesure de composant biologique dans laquelle un composé d'aminoantipyrine est utilisé en tant que coupleur d'un réactif de coloration redox, l'agent pour supprimer une réduction de la sensibilité de mesure d'un composant biologique étant caractérisé en ce qu'il contient une substance contenant du cuivre. De préférence, la méthode de mesure de composant biologique implique l'utilisation d'un réactif ou d'un ensemble de réactifs qui satisfait les exigences (a) à (d) ci-dessous, la méthode étant caractérisée en ce que la substance contenant du cuivre est utilisée en étant ajoutée à un réactif qui satisfait l'exigence (d) : (a) contient une oxydase capable de générer du peroxyde d'hydrogène. (b) contient une peroxydase. (c) contient un réactif de coloration redox qui se colore lors de la réaction avec du peroxyde d'hydrogène en présence de peroxydase. (d) contient un composé aminoantipyrine en tant que coupleur du réactif de coloration redox.
PCT/JP2019/018704 2018-05-10 2019-05-10 Méthode de suppression de la réduction de la sensibilité d'un kit de réactif de mesure de composant biologique WO2019216405A1 (fr)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS60262599A (ja) * 1984-06-09 1985-12-25 Wako Pure Chem Ind Ltd アスコルビン酸の新規な分解方法
JPH07303497A (ja) * 1994-05-13 1995-11-21 Toyobo Co Ltd 生体成分の測定方法および測定用試薬組成物
JPH11504808A (ja) * 1995-05-05 1999-05-11 ジェンザイム・リミテッド 糖化タンパク質の測定
JP2016019497A (ja) * 2014-07-15 2016-02-04 東洋紡株式会社 生体成分の測定方法および測定用組成物

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS60262599A (ja) * 1984-06-09 1985-12-25 Wako Pure Chem Ind Ltd アスコルビン酸の新規な分解方法
JPH07303497A (ja) * 1994-05-13 1995-11-21 Toyobo Co Ltd 生体成分の測定方法および測定用試薬組成物
JPH11504808A (ja) * 1995-05-05 1999-05-11 ジェンザイム・リミテッド 糖化タンパク質の測定
JP2016019497A (ja) * 2014-07-15 2016-02-04 東洋紡株式会社 生体成分の測定方法および測定用組成物

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
KASAHARA, SHIZUKA ET AL.,: "Redox color reagents for the assay of hydroperoxides", KAGAKU, vol. 28, 1979, pages 405 - 410, XP055651714 *

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