Reagent for biochemical testing and method for producing same

Abstract

The present disclosure provides a reagent for biochemical testing that exhibits excellent long-term storage stability. The present disclosure also provides a method for suitably producing such a reagent for biochemical testing. The reagent according to the present disclosure includes heated catalase and is used in biochemical testing.

Description

Reagent for Biochemical Examination and Method for Producing the Same

[0009]

[0001] The present disclosure relates to a reagent for biochemical examination and a method for producing the same.

[0002] Biochemical examination is an examination that analyzes various components contained in various biological samples such as collected blood and urine, and estimates, for example, the presence or absence of abnormalities, disease sites, the presence or absence of inflammation, nutritional status, etc. A reagent used for such an examination is called a reagent for biochemical examination, and various reagent components are included according to the examination items. Reagents for biochemical examination are a type of clinical examination drug, and among the reagents for biochemical examination, there are also reagents to which catalase is added in order to eliminate components that interfere with measurement or to stabilize reagent components.

[0003] For example, in Patent Document 1, a reagent for triglyceride measurement containing a specific catalase and adenosine triphosphate has been proposed.

[0004] Japanese Patent No. 7571418

[0005] However, the present inventors have found that there are problems in conventional reagents containing catalase. That is, in conventional reagents, catalase in the reagent may be inactivated during long-term storage. As a result, the original function of catalase may not be fully exerted, and there is a risk of affecting the measurement system. Such concerns exist even when using high-grade products of commercially available catalase that are used as raw materials for clinical examination drugs.

[0006] ​​​​​​​​​​

[0010] Disclosure 4 is a reagent used in an enzymatic method, one of the reagents described in Disclosures 1 to 3.

[0011] Disclosure 5 is a reagent from any one of Disclosures 1 to 3 used in a non-enzymatic method.

[0012] Disclosure 6 is one of the reagents from Disclosures 1 to 5, wherein the target of measurement for the biochemical test described above is a biomolecule.

[0013] Disclosure 7 is a reagent according to any one of Disclosures 1 to 6, wherein the biomolecule is at least one selected from the group consisting of sugars and their metabolites, lipids and their metabolites, proteins, enzymes, hormones and their precursors, and nitrogen-containing components.

[0014] Disclosure 8 is a reagent from any one of Disclosures 1 to 7, wherein the biomolecule is at least one selected from the group consisting of glucose, triglycerides, albumin, glycated albumin, hemoglobin, glycated hemoglobin, globulin, cholesterol, uric acid, creatinine, polyamine, bile acid, 1,5-anhydroglucitol, pyruvate, lactate, phospholipids, urea, creatine, free fatty acids, human brain natriuretic peptide (BNP), insulin, procalcitonin (PCT), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK), alkaline phosphatase (ALP), lactate dehydrogenase (LD), γ-glutamyltransferase (γ-GT), amylase, cholinesterase (ChE), and amylase isozymes.

[0015] Disclosure 9 is one of the reagents from Disclosures 1 to 8, wherein the ratio (X / Y) of the concentration ratio X calculated by the following formula (1) to the concentration ratio Y calculated by the following formula (2) is 100% or more. X = (A1 / B1) × 100 (1) Y = (A2 / B2) × 100 (2) The symbols in the formulas are as follows: A1: Catalase concentration in the reagent after storing the reagent at 37°C for 24 hours (U / mL) B1: Catalase concentration in the reagent after storing the reagent at 4°C for 24 hours (U / mL) A2: Catalase concentration in the reagent y, which is the same as the reagent above except that the catalase is not heated, after storing it at 37°C for 24 hours (U / mL) B2: Catalase concentration in the reagent y after storing it at 4°C for 24 hours (U / mL)

[0016] Disclosure 10 is a method for producing a reagent used in biochemical testing, comprising a reagent component and catalase, and the method includes a contact step of bringing the catalase into contact with the reagent component while it is heated.

[0017] Disclosure 11 is a method for producing Disclosure 10, wherein the contact step is one of the following (a) to (c): (a) a step of heating catalase and a step of mixing the heated catalase with a reagent component; (b) a step of mixing catalase and a reagent component under heating; (c) a step of mixing catalase and a reagent component and a step of heating the mixture obtained by the mixing step.

[0018] Disclosure 12 is a method for producing the product according to Disclosure 10 or 11, wherein the contact step includes a step of heating the catalase and a step of mixing the heated catalase with the reagent component.

[0019] Disclosure 13 is a reagent from any one of Disclosures 1 to 9, obtained by the manufacturing method of Disclosure 10, 11, or 12.

[0020] This disclosure provides a biochemical testing reagent with excellent long-term storage stability. Furthermore, this disclosure also provides a suitable method for obtaining such a biochemical testing reagent.

[0021] [Reagents] The reagents of this disclosure are for biochemical testing and include catalase after heating.

[0022] Catalase is generally produced by living organisms such as animals, plants, and microorganisms. In this disclosure, catalase is preferably of microbial or animal origin. The type of microorganism is not particularly limited, but examples of microorganisms include Aspergillus, Micrococcus, Bacillus subtilis, Thermomyces, Bacillus sp., Acremonium alabamensis, Thermoascus aurantiacu, Alkaligenes deleya, and Alkaligenes microcilla. Examples of Aspergillus species include Aspergillus niger, Aspergillus carbonarius, and Aspergillus terreus. Among these, microbial catalase is preferably derived from Aspergillus or Micrococcus species. Animal-derived catalase is preferably derived from bovine liver. In this disclosure, catalase may be obtained by isolating and extracting catalase from the cells or tissues of the organism, or by extracting and purifying catalase from a culture obtained by culturing the cells, tissues, or genetically modified cells of catalase derived from the organism, or by using a commercially available product.

[0023] The above reagent contains catalase after heating. Catalase after heating is obtained by heating catalase. That is, the above reagent contains catalase that has been heated.

[0024] The heating temperature should be such that the catalase can exert its catalytic effect without being completely deactivated after heating. The specific heating temperature will vary depending on the type of catalase and the operating temperature range, but as will be described later in the reagent manufacturing method, it is preferably 30 to 70°C, and more preferably 55 to 65°C. The heating time is also preferably 2 to 350 hours, more preferably 5 to 320 hours, and even more preferably 10 to 300 hours, as will be described later in the reagent manufacturing method.

[0025] It is preferable that the above reagent has a ratio (X / Y) of 100% or more between the concentration ratio X calculated by the following formula (1) and the concentration ratio Y calculated by the following formula (2). X = (A1 / B1) × 100 (1) Y = (A2 / B2) × 100 (2) The symbols in the formulas are as follows: A1: Catalase concentration in the reagent after storing the reagent at 37°C for 24 hours (U / mL) B1: Catalase concentration in the reagent after storing the reagent at 4°C for 24 hours (U / mL) A2: Catalase concentration in the reagent after storing reagent y, which is the same as the above reagent except that the catalase is not heated, at 37°C for 24 hours (U / mL) B2: Catalase concentration in the reagent after storing reagent y at 4°C for 24 hours (U / mL)

[0026] Biochemical test reagents are typically stored and used at low temperatures (e.g., 4°C). Therefore, storing a reagent at 37°C for 24 hours after preparation is equivalent to an accelerated test under normal storage conditions. Specifically, a test of storing a reagent at 37°C for 24 hours may be equivalent to a test of storing a reagent at 4°C for several months (e.g., 1-2 months). Normally, if a reagent is stored at a temperature exceeding the recommended storage temperature, the components may react over time, and in the case of reagents containing catalase, the catalase concentration may decrease. Therefore, the higher the ratio of the catalase concentration A when a reagent is stored at 37°C for 24 hours after preparation to the catalase concentration B when the reagent is stored at 4°C for 24 hours after preparation (A / B; also called the recovery rate), the better the long-term storage stability. In particular, the higher the ratio of the recovery rate X of a reagent to the recovery rate Y of an unheated catalase-containing reagent y (i.e., X / Y), the better the long-term storage stability of that reagent. Reagent y is a reagent having the same composition as the reagent in this disclosure, except that the catalase is not heated.

[0027] When the above ratio (X / Y) is 100% or more, the reagent exhibits excellent long-term storage stability and is therefore more effective as a reagent for biochemical testing. It is more preferable that the above ratio (X / Y) exceeds 100%, and even more preferable that it is 103% or more. Furthermore, from the viewpoint of excellent long-term storage stability, it is preferable that the concentration ratio X calculated by the above formula (1) is 90% or more, and more preferable that it is 94% or more.

[0028] In biochemical tests using the above reagents, for example, a biological sample is used as the specimen, and various components contained in the biological sample are analyzed (measured). Biological samples include, for example, tissue cells, serum, plasma, blood cells, urine, feces, saliva, pancreatic juice, and other bodily fluids or various secretions collected from a living organism.

[0029] The target of the above biochemical tests is preferably a biomolecule. A biomolecule is a compound in a living organism that has a function related to life phenomena. A biomolecule may also be a complex of biomolecules.

[0030] The above biomolecules are preferably at least one selected from the group consisting of sugars and their metabolites, lipids and their metabolites, proteins, enzymes, hormones and their precursors, and nitrogen-containing components. The reagents of this disclosure are suitably applied to such biomolecules.

[0031] Examples of the sugars and their metabolites include glucose, glycated albumin, glycated hemoglobin, 1,5-anhydroglucitol, pyruvate, and lactic acid. Examples of the lipids and their metabolites include triglycerides, cholesterol, bile acids, phospholipids, and free fatty acids. Examples of the proteins include albumin, hemoglobin, and globulin. Examples of the enzymes include reagents that can be used in enzyme calibrators, specifically, aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK), alkaline phosphatase (ALP), lactate dehydrogenase (LD), γ-glutamyltransferase (γ-GT), amylase, cholinesterase (ChE), and amylase isozymes. Examples of the hormones and their precursors include human brain natriuretic peptide (BNP), insulin, and procalcitonin (PCT). Examples of nitrogen-containing components include uric acid, creatinine, polyamines, urea, and creatine.

[0032] The above biomolecules are preferably at least one selected from the group consisting of glucose, triglycerides, albumin, glycated albumin, hemoglobin, glycated hemoglobin, globulin, cholesterol, uric acid, creatinine, polyamines, bile acids, 1,5-anhydroglucitol, pyruvate, lactate, phospholipids, urea, creatine, free fatty acids, human brain natriuretic peptide (BNP), insulin, procalcitonin (PCT), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK), alkaline phosphatase (ALP), lactate dehydrogenase (LD), γ-glutamyltransferase (γ-GT), amylase, cholinesterase (ChE), and amylase isozymes. Among these, for example, glucose and glycated hemoglobin are examples of test items for diabetes, triglycerides are an example of a test item for arteriosclerosis, and albumin is an example of a test item for nutritional status.

[0033] The above reagent contains various reagent components corresponding to the test items (measurements) of the biochemical test. That is, the above reagent includes catalase after heating and reagent components. Reagent components refer to components involved in the reaction system (e.g., enzymes, substrates, antibodies, labeling substances, colorants, etc.) and / or components not involved in the reaction system (e.g., stabilizers, preservatives, emulsifiers, solvents, buffers, excipients, etc.). Hereinafter, components involved in the reaction system will also be referred to as "main components," and components not involved in the reaction system will also be referred to as "auxiliary components." For example, commercially available in vitro diagnostic medical devices include products composed of main components and auxiliary components, and products composed of only main components or only auxiliary components. In this specification, "reagent components" (i.e., main components and / or auxiliary components) do not include catalase after heating.

[0034] Of the total amount of the above reagents, the combined amount of heated catalase and reagent components is preferably 95% by mass or more, and more preferably 100% by mass. The above reagents may contain unheated catalase, but in this case, the content of unheated catalase is preferably 3% by mass or less, more preferably 1% by mass or less, of the total amount of the above reagents, and even more preferably substantially free of unheated catalase.

[0035] In the above reagent, the catalase content after heating is preferably 50 U / mL to 20000 U / mL at the final concentration in the reagent, more preferably 50 U / mL to 1000 U / mL, even more preferably 50 U / mL to 500 U / mL, particularly preferably 100 U / mL to 500 U / mL, and most preferably 100 U / mL to 200 U / mL.

[0036] In the reagents described above, the ratio of the main component to the auxiliary component varies depending on the type of each component, but for example, their mass ratio (main component / auxiliary component) may be 0-50 / 50-100 or 0.01-50 / 50-99.9.

[0037] The reagents described above are preferably reagents used in enzymatic methods. Enzymatic methods are techniques for quantifying target substances using enzymatic reactions. The reagents described above are also preferably reagents used in non-enzymatic methods. Non-enzymatic methods are biochemical testing techniques other than enzymatic methods, and include, for example, colorimetric methods; immunoassays (e.g., latex immunoturbidimetry); electrophoresis; fluorescence measurement; chromatography (e.g., high-performance liquid chromatography (HPLC)); and coagulation time methods.

[0038] The use of heated catalase as a reagent for biochemical testing (i.e., application of heated catalase to biochemical testing reagents) is included in this disclosure.

[0039] [Method for Manufacturing Reagents] The method for manufacturing reagents used in biochemical tests comprises a reagent component and catalase, and includes a contact step of bringing the catalase into contact with the reagent component while it is heated. The reagents of the present disclosure described above can be suitably obtained by such a manufacturing method.

[0040] The above contact step is preferably one of the following (a) to (c): (a) A step comprising heating catalase (a1) and mixing the heated catalase with the reagent component (a2); (b) A step of mixing catalase and the reagent component under heating; (c) A step comprising mixing catalase and the reagent component (c1) and heating the mixture obtained in the mixing step (c1) (c2).

[0041] The catalase used in steps (a1), (b), and (c1) described above is of biological origin, such as animals, plants, and microorganisms, and is preferably of microbial or animal origin, as described above. The preferred embodiment of the catalase is as described above.

[0042] The reagent components used in steps (a2), (b), and (c1) above may be those corresponding to the test items (measurements) of the biochemical test. The reagent components are as described above.

[0043] In step (a2) above, the amount of catalase added after heating is preferably 50 U / mL to 300,000 U / mL at a final concentration, more preferably 50 U / mL to 200,000 U / mL, even more preferably 50 U / mL to 100,000 U / mL, and most preferably 100 U / mL to 100,000 U / mL. Also, in steps (b) and (c1) above, the amount of catalase added is preferably 50 U / mL to 300,000 U / mL at a final concentration, more preferably 50 U / mL to 200,000 U / mL, even more preferably 50 U / mL to 100,000 U / mL, and most preferably 100 U / mL to 100,000 U / mL.

[0044] In the above steps (a1), (b), and (c2), the heating temperature and heating time of catalase can be appropriately adjusted within the range where the catalytic effect of the catalase can be exerted depending on the catalase used, such as short time at high temperature or long time at low temperature. Also, the heating temperature may be any temperature at which the catalase after heating can exhibit a catalytic effect without being completely inactivated. Specific heating temperatures vary depending on the type of catalase, the operating temperature range, etc., but for example, it is preferably 30 to 70°C, more preferably 55 to 65°C. Also, the heating time is preferably, for example, 2 to 350 hours, more preferably 5 to 320 hours, and even more preferably 10 to 300 hours.

[0045] The above production method may include other steps other than the above contact step. For example, the above production method may include a step of cooling the system (for example, a step of cooling the reaction product or mixture at that time) during or after the above contact step. The cooling step is preferably a step of cooling the system to a temperature below the heating temperature in the above contact step. For example, it is preferably to cool the reaction product or mixture at that time to 10°C or lower, and more preferably to 5°C or lower. When the above production method further includes such a cooling step, the resulting reagent will be even more excellent in long-term storage stability.

[0046] The above contact step is particularly preferably the above step (a). That is, the above contact step preferably includes a step (a1) of heating catalase and a step (a2) of mixing the heated catalase with the reagent components. In this case, the reagent obtained by the above production method will be even more excellent in long-term storage stability.

[0047] The above step (a) may further include a step (a3) of cooling the heated catalase between the above step (a1) and the above step (a2). The step (a3) is a step of cooling the heated catalase to a temperature below the heating temperature. For example, it is preferably cooled to 10°C or lower, and more preferably to 5°C or lower. When the above step (a) includes the above step (a1), the above step (a3), and the above step (a2) in this order, the resulting reagent will be even more excellent in long-term storage stability.

[0048] The above step (a2) is preferably carried out, for example, in an environment at room temperature (for example, 1 to 30°C). In this case, the catalase after heating and the reagent components can be more efficiently mixed while further suppressing the decrease in the catalase concentration.

[0049] The present disclosure will be described in more detail below with specific examples, but the present disclosure is not limited to only these examples. Unless otherwise specified, "%" means "mass %".

[0050] 1. Materials In each of the test examples described below, the following catalase and reagent components were used. (1) Catalase: Any one of catalases C1 to C3 shown in Table 1 was used. The properties of each catalase shown in Table 1 are the properties when stored at the storage temperature recommended by each manufacturer (that is, C1 and C2 are stored at 4°C, and C3 is stored at -20°C). However, even at a temperature exceeding the storage temperature (for example, room temperature or the heating temperature of the catalase), each catalase shows the same properties as those in Table 1. (2) Reagent components: Any one of reagent components R1 to R4 having the compositions shown in Table 2 was used. Table 2 shows the final concentrations of the contained components (that is, the materials before mixing) contained in each component.

[0051]

[0052]

[0053] 2. Temperature measurement For the temperature measurement and recording of the catalase, "testo Saveris" ver. 1 (37°C) and ver. 3 (60°C) manufactured by Testo SE & Co. KGaA were used.

[0054] 3. Measurement of absorbance (1) After preparing the reagent according to the test example described later, 10 μL of the reagent was diluted with 90 μL of diluent to obtain a diluted sample. Sodium phosphate buffer (final concentration: 50 mM) was used as the diluent. 62 μL of analytical reagent Z1 shown in Table 3 was added to 2.0 μL of the obtained diluted sample, and after holding at 37°C for 5 minutes, 24 μL of analytical reagent Z2 shown in Table 3 was added to obtain the measurement sample. The absorbance of the measurement sample 5 minutes after the addition of analytical reagent Z2 was measured at 805 nm / 545 nm according to (2) below. The difference in absorbance obtained, i.e., the value obtained by subtracting the absorbance at the secondary wavelength of 805 nm from the absorbance at the primary wavelength of 545 nm, was defined as absorbance Abs. Using this absorbance Abs, the catalase concentration was calculated according to (3) below. Table 3 shows the components and final concentrations of analytical reagents Z1 and Z2.

[0055]

[0056] (2) Absorbance measurements at 805 nm and 545 nm were performed using the JCA-BM series automated analyzer cleaner (JCA-BM9130) manufactured by JEOL Ltd. under the following measurement conditions: • Analytical method: EPA (endpoint method) • Reaction time: 10 minutes • Measurement points: 44, 47-49, 98 • Measurement wavelength (secondary / primary): 805 nm / 545 nm

[0057] (3) As a method for calculating the catalase concentration from the absorbance Abs, a calibration curve was prepared in advance using a catalase standard solution with known activity values, and the catalase concentration in the sample was determined using this calibration curve. That is, it was determined as follows: First, the concentration (U / mL) of the calibration sample (catalase standard solution) was plotted on the x-axis and the absorbance Abs of the sample was plotted on the y-axis, and a linear regression equation was calculated. Next, the catalase concentration was calculated using the following formula.

[0058]

[0059] 4. Preparation and Evaluation of Reagents (Test Example 1-1) After adding reagent component R1 to a reaction vessel, catalase C1, which had been stored at 4°C, was added to the vessel without heating. The contents of the vessel were stirred and mixed at room temperature (1-30°C) to prepare the reagent. The prepared reagent was dispensed into two containers, one was stored at 4°C for 24 hours, and the other was stored at 37°C for 24 hours. The absorbance of each reagent was then measured according to section 3 above, and the catalase concentration in each reagent was calculated. The catalase concentration B2 in the reagent after storage at 4°C is recorded in column B of Table 4, the catalase concentration A2 in the reagent after storage at 37°C is recorded in column A of Table 4, and the ratio of these concentrations Y (A2 / B2) is recorded in column Y of Table 4.

[0060] (Test Example 1-2) Catalase C1, which had been stored at 4°C, was placed in a heating container and heated at 37°C for 12 hours, then cooled to 4°C. Next, this catalase C1 was placed in a reaction vessel containing reagent component R1, and the contents of the container were stirred and mixed at room temperature (1-30°C) to prepare the reagent. The prepared reagent was dispensed into two containers, one was stored at 4°C for 24 hours, and the other was stored at 37°C for 24 hours. The absorbance of each reagent was then measured according to 3. above, and the catalase concentration in each reagent was calculated. The catalase concentration B1 in the reagent after storage at 4°C is recorded in column B of Table 4, the catalase concentration A1 in the reagent after storage at 37°C is recorded in column A of Table 4, and the ratio of these concentrations X (A1 / B1) is recorded in column X of Table 4. Table 4 shows the value (X / Y) obtained by dividing the concentration ratio X (A1 / B1) obtained in this example by the concentration ratio Y (A2 / B2) obtained in Test Example 1-1.

[0061] (Test Example 1-3) Catalase C1, which had been stored at 4°C, was placed in a heating container and heated at 63°C for 12 hours, then cooled to 4°C. Next, this catalase C1 was placed in a reaction vessel containing reagent component R1, and the contents of the container were stirred and mixed at room temperature (1-30°C) to prepare the reagent. The prepared reagent was dispensed into two containers, one was stored at 4°C for 24 hours, and the other was stored at 37°C for 24 hours. The absorbance of each reagent was then measured according to 3. above, and the catalase concentration in each reagent was calculated. The catalase concentration B1 in the reagent after storage at 4°C is recorded in column B of Table 4, the catalase concentration A1 in the reagent after storage at 37°C is recorded in column A of Table 4, and the ratio of these concentrations X (A1 / B1) is recorded in column X of Table 4. Table 4 shows the value (X / Y) obtained by dividing the concentration ratio X (A1 / B1) obtained in this example by the concentration ratio Y (A2 / B2) obtained in Test Example 1-1.

[0062]

[0063] (Test Example 2-1) The concentration ratio Y (A2 / B2) was calculated in the same manner as in Test Example 1-1, except that reagent component R2 was used instead of reagent component R1 (see column Y in Table 5).

[0064] (Test Example 2-2) The concentration ratio X (A1 / B1) was calculated in the same manner as in Test Example 1-2, except that reagent component R2 was used instead of reagent component R1 (see column X in Table 5). The value (X / Y) obtained by dividing the concentration ratio X (A1 / B1) obtained in this example by the concentration ratio Y (A2 / B2) obtained in Test Example 2-1 is shown in Table 5.

[0065] (Test Example 2-3) The concentration ratio X(A1 / B1) was calculated in the same manner as in Test Example 1-3, except that reagent component R2 was used instead of reagent component R1 (see column X in Table 5). The value (X / Y) obtained by dividing the concentration ratio X(A1 / B1) obtained in this example by the concentration ratio Y(A2 / B2) obtained in Test Example 2-1 is shown in Table 5.

[0066] (Test Example 2-4) The concentration ratio Y (A2 / B2) was calculated in the same manner as in Test Example 2-1, except that catalase C2, which was stored at 4°C, was used instead of catalase C1, which was stored at 4°C (see column Y in Table 5).

[0067] (Test Example 2-5) The concentration ratio X (A1 / B1) was calculated in the same manner as in Test Example 2-2, except that catalase C2, which was stored at 4°C, was used instead of catalase C1, which was stored at 4°C (see column X in Table 5). The value (X / Y) obtained by dividing the concentration ratio X (A1 / B1) obtained in this example by the concentration ratio Y (A2 / B2) obtained in Test Example 2-4 is shown in Table 5.

[0068] (Test Example 2-6) The concentration ratio Y (A2 / B2) was calculated in the same manner as in Test Example 2-1, except that catalase C3, which had been stored at -20°C, was used instead of catalase C1, which had been stored at 4°C (see column Y in Table 5).

[0069] (Test Example 2-7) The concentration ratio X (A1 / B1) was calculated in the same manner as in Test Example 2-2, except that catalase C3, which had been stored at -20°C, was used instead of catalase C1, which had been stored at 4°C (see column X in Table 5). The value (X / Y) obtained by dividing the concentration ratio X (A1 / B1) obtained in this example by the concentration ratio Y (A2 / B2) obtained in Test Example 2-6 is shown in Table 5.

[0070]

[0071] (Test Example 3-1) The concentration ratio Y (A2 / B2) was calculated in the same manner as in Test Example 1-1, except that reagent component R3 was used instead of reagent component R1 (see column Y in Table 6).

[0072] (Test Example 3-2) The concentration ratio X(A1 / B1) was calculated in the same manner as in Test Example 1-3, except that reagent component R3 was used instead of reagent component R1 (see column X in Table 6). The value (X / Y) obtained by dividing the concentration ratio X(A1 / B1) obtained in this example by the concentration ratio Y(A2 / B2) obtained in Test Example 3-1 is shown in Table 6.

[0073]

[0074] (Test Example 4-1) The concentration ratio Y (A2 / B2) was calculated in the same manner as in Test Example 1-1, except that reagent component R4 was used instead of reagent component R1 (see column Y in Table 7).

[0075] (Test Example 4-2) The concentration ratio X (A1 / B1) was calculated in the same manner as in Test Example 1-2, except that reagent component R4 was used instead of reagent component R1 (see column X in Table 7). The value (X / Y) obtained by dividing the concentration ratio X (A1 / B1) obtained in this example by the concentration ratio Y (A2 / B2) obtained in Test Example 4-1 is shown in Table 7.

[0076] (Test Example 4-3) The concentration ratio X (A1 / B1) was calculated in the same manner as in Test Example 1-3, except that reagent component R4 was used instead of reagent component R1 (see column X in Table 7). The value (X / Y) obtained by dividing the concentration ratio X (A1 / B1) obtained in this example by the concentration ratio Y (A2 / B2) obtained in Test Example 4-1 is shown in Table 7.

[0077] (Test Example 4-4) The concentration ratio Y (A2 / B2) was calculated in the same manner as in Test Example 4-1, except that catalase C2, which was stored at 4°C, was used instead of catalase C1, which was stored at 4°C (see column Y in Table 7).

[0078] (Test Example 4-5) The concentration ratio X (A1 / B1) was calculated in the same manner as in Test Example 4-2, except that catalase C2, which was stored at 4°C, was used instead of catalase C1, which was stored at 4°C (see column X in Table 7). The value (X / Y) obtained by dividing the concentration ratio X (A1 / B1) obtained in this example by the concentration ratio Y (A2 / B2) obtained in Test Example 4-4 is shown in Table 7.

[0079] (Test Example 4-6) The concentration ratio Y (A2 / B2) was calculated in the same manner as in Test Example 4-1, except that catalase C3 stored at -20°C was used instead of catalase C1 stored at 4°C (see column Y in Table 7).

[0080] (Test Example 4-7) The concentration ratio X (A1 / B1) was calculated in the same manner as in Test Example 4-2, except that catalase C3, which had been stored at -20°C, was used instead of catalase C1, which had been stored at 4°C (see column X in Table 7). The value (X / Y) obtained by dividing the concentration ratio X (A1 / B1) obtained in this example by the concentration ratio Y (A2 / B2) obtained in Test Example 4-6 is shown in Table 7.

[0081]

[0082] 5. From Analysis Tables 4 to 7, it can be confirmed that the catalase concentration ratio (A / B) was significantly improved in the reagent containing heated catalase compared to the reagent containing unheated catalase. Furthermore, the concentration ratio X of the reagent containing heated catalase was higher than the concentration ratio Y of the reagent containing unheated catalase, and these ratios (X / Y) exceeded 100%, specifically 103% or more. Therefore, it was found that the reagent of this disclosure has excellent long-term storage stability due to the inclusion of heated catalase. According to the inventors' investigation, the reason for this phenomenon is thought to be that the molecular structure of catalase was stabilized by heating.

Claims

1. A reagent containing heated catalase, used in biochemical tests.

2. The reagent according to claim 1, wherein the catalase is derived from a microorganism or an animal.

3. The reagent according to claim 1 or 2, wherein the catalase is derived from Aspergillus, Micrococcus, or bovine liver.

4. A reagent used in an enzymatic method, according to any one of claims 1 to 3.

5. A reagent according to any one of claims 1 to 3, used in a non-enzymatic method.

6. The reagent according to any one of claims 1 to 5, wherein the target of measurement for the biochemical test is a biomolecule.

7. The reagent according to claim 6, wherein the biomolecule is at least one selected from the group consisting of sugars and their metabolites, lipids and their metabolites, proteins, enzymes, hormones and their precursors, and nitrogen-containing components.

8. The reagent according to claim 6, wherein the biomolecule is at least one selected from the group consisting of glucose, triglycerides, albumin, glycated albumin, hemoglobin, glycated hemoglobin, globulin, cholesterol, uric acid, creatinine, polyamine, bile acid, 1,5-anhydroglucitol, pyruvate, lactate, phospholipids, urea, creatine, free fatty acids, human brain natriuretic peptide (BNP), insulin, procalcitonin (PCT), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK), alkaline phosphatase (ALP), lactate dehydrogenase (LD), γ-glutamyltransferase (γ-GT), amylase, cholinesterase (ChE), and amylase isozymes.

9. The reagent according to any one of claims 1 to 8, wherein the ratio (X / Y) of the concentration ratio X calculated by the following formula (1) to the concentration ratio Y calculated by the following formula (2) is 100% or more. X = (A1 / B1) × 100 (1) Y = (A2 / B2) × 100 (2) The symbols in the formulas are as follows: A1: Catalase concentration in the reagent after storing the reagent at 37°C for 24 hours (U / mL) B1: Catalase concentration in the reagent after storing the reagent at 4°C for 24 hours (U / mL) A2: Catalase concentration in the reagent after storing reagent y, which is the same as the reagent except that the catalase is not heated, at 37°C for 24 hours (U / mL) B2: Catalase concentration in the reagent after storing reagent y at 4°C for 24 hours (U / mL) 10. A method for producing a reagent used in biochemical testing, comprising a reagent component and catalase, the method comprising a contact step of bringing the catalase into contact with the reagent component while it is heated.

11. The manufacturing method according to claim 10, wherein the contact step is any of the following (a) to (c): (a) a step of heating catalase and a step of mixing the heated catalase with a reagent component; (b) a step of mixing catalase and a reagent component under heating; (c) a step of mixing catalase and a reagent component and a step of heating the mixture obtained by the mixing step.

12. The manufacturing method according to claim 10, wherein the contact step includes the steps of heating the catalase and mixing the heated catalase with the reagent component.

13. The reagent according to any one of claims 1 to 9, obtained by the manufacturing method described in claim 10, 11, or 12.

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