JPH07303497A - Assay of biological component and reagent composition therefor - Google Patents

Abstract

PURPOSE:To accurately determine a biological component without interference of ascorbic acid, by making ascorbic acid oxidase and an acid manganese salt act on a biological component etc., in a specimen to remove inhibitory substance(s) in the component followed by making an oxidase act on the component and quantifying the H2O2 formed. CONSTITUTION:Ascorbic acid oxidase and an organic or inorganic acid manganese salt (e.g. manganese acetate) are made to act on a biological component (e.g. glucose) in a specimen such as serum or urine or a substance derived from this component to remove inhibitory substance(s) in the component followed by making an oxidase (e.g. glucose oxidase) act on this component, and the hydrogen peroxide formed is subjected to colorimetry through formation of a color by using peroxidase, a chromogen [e.g. N-ethyl-N-(2-hydroxy-3- sulfopropyl)-3,5-dimethylaniline and a coupler (e.g. 4-aminoantipyrine) to exclude the error due to ascorbic acid, thus determining the biological component in the specimen accurately.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for measuring biological components and a reagent composition for measurement. More specifically, the method for measuring a biological component using an oxidase and the reagent composition for the measurement enable accurate measurement without error due to ascorbic acid.

[0002]

2. Description of the Related Art The presence of components such as glucose, cholesterol and uric acid in biological samples such as serum and urine and the determination of their amounts provide useful information in the field of clinical examination. As the measuring method, an enzymatic measuring method using an enzyme having high substrate specificity for each component is rapidly prevailing. Among them, a method in which an oxidase is allowed to act on a biological component to be measured and the produced hydrogen peroxide is measured is often used. Examples of a combination of a representative component and an oxidase that directly acts on the component include glucose and glucose oxidase, free cholesterol and cholesterol oxidase, and uric acid and uricase.

When the oxidase cannot act directly on the biological component to be measured, the enzyme present in the biological component is allowed to act in advance to generate a substance serving as a substrate for the oxidase, and then the oxidase is added to the substance. Is used to measure the hydrogen peroxide produced. Examples of such representatives include neutral fat and glycerol-3-phosphate oxidase, inorganic phosphorus and xanthine oxidase, phospholipid and choline oxidase, and creatinine and sarcosine oxidase.

Hydrogen peroxide produced by the action of an oxidase can be measured by various known methods. The most commonly used method is to oxidize a chromogen in the presence of peroxidase. Examples include a method in which a dye is formed by condensation with a coupler such as 4-aminoantipyrine to perform colorimetric determination. Chromogens used in this method include phenol derivatives such as phenol, 2-chlorophenol, 4-chlorophenol, and 2,4-dichlorophenol, or aniline, N, N-dimethylaniline, N, N-diethylaniline. , N, N-diethyl-m
-Toluidine, N, N-diethyl-m-anisidine, N
-Ethyl-N- (3-methylphenyl) -N'-acetylethylenediamine, N-ethyl-N- (2-hydroxy-3-sulfopropyl) -m-anisidine, N-ethyl-N- (2-hydroxy- 3-sulfopropyl) -m
-Toluidine, N-ethyl-N-sulfopropyl-m-
Examples include aniline derivatives such as toluidine and N-ethyl-N-sulfopropyl-m-anisidine.

[0005]

In such a method for measuring a biological component, it is often experienced that a reducing substance contained in a biological sample, particularly ascorbic acid, causes a negative error in the measurement and a correct measurement result cannot be obtained. Has been done.
As a means for solving this problem, a method has been used in which ascorbic acid oxidase is allowed to coexist in a reagent to efficiently decompose ascorbic acid and avoid interference with the measurement of biological components. However, since ascorbic acid oxidase is easily deactivated in a solution, there is a drawback that the resolution of ascorbic acid is lowered during storage of the reagent. As a means for preventing a decrease in the resolution of ascorbic acid, various stabilizers have been added to ascorbate oxidase. Examples of stabilizers include
Examples thereof include boric acid and sucrose. However,
Although these methods improved the stability of ascorbate oxidase in solution, they did not completely avoid interference with ascorbate measurements.

[0006]

DISCLOSURE OF THE INVENTION The inventors of the present invention have made earnest studies on a method for avoiding interference with the measured value of ascorbic acid in view of the above-mentioned situation. As a result, ascorbic acid oxidase and a manganese salt of an organic acid or an inorganic acid are selected. By coexisting, the above-mentioned subject was solved.

That is, the present invention provides a method for measuring a biological component by causing an oxidase to act on a biological component in a sample or a substance derived from the biological component, and measuring the produced hydrogen peroxide. A method for measuring a biological component, which is characterized in that a manganese salt of an acid or an inorganic acid is allowed to act to remove an interfering substance in the biological component.

The present invention also provides a reagent composition for measuring biological components, which comprises an oxidase, a peroxidase, a chromogen, a coupler, an ascorbate oxidase and a manganese salt of an organic acid or an inorganic acid.

Further, the present invention is characterized by containing an enzyme that produces a substrate for an oxidase from a biological component, an oxidase, a peroxidase, a chromogen, a coupler, an ascorbate oxidase, and a manganese salt of an organic acid or an inorganic acid. It is a reagent composition for measuring components.

The biological components to be measured in the present invention include glucose, free cholesterol, ester type cholesterol, neutral fat, phospholipid, inorganic phosphorus, free fatty acid, urea nitrogen, pyruvic acid, creatine, creatinine or uric acid. . The components to be measured in the present invention are not limited to these components, but include components that can be measured by causing oxidase to act to generate hydrogen peroxide and measuring the amount thereof.

The substances derived from biological components to be measured in the present invention include glycerol or glycerol-3-phosphate derived from neutral fat, choline derived from phospholipids, creatine, sarcosine derived from creatinine, α -Glucose derived from amylase. The components to be measured in the present invention also include enzymes such as α-amylase. The component to be measured in the present invention is not limited to these components, and it is a substance that produces a substance that serves as a substrate for an oxidase by, for example, a chemical reaction, and the oxidase acts on the substance to produce hydrogen peroxide. It also includes components that can be measured by measuring the amount thereof.

Examples of oxidases that directly act on biological components include glucose oxidase, cholesterol oxidase, glycerol oxidase, xanthine oxidase, pyruvate oxidase, uricase and the like.

As an enzyme for obtaining a substance serving as a substrate for oxidase from a biological component, lipoprotein lipase, glycerol kinase, phospholipase D, cholesterol esterase, purine nucleoside phosphorylase, creatine amidinohydrolase, creatinine amide hydrolase, acyl CoA Examples include synthetase and urea amide lyase.

The biological components to be measured in the present invention, the enzyme for producing a substrate for oxidase from the biological components, and the oxidase used are summarized below.

[0015]

[Table 1]

The couplers used in the present invention include 4-
Aminoantipyrine, 3-methyl-2-benzothiazolinone hydrazone and the like can be mentioned.

Chromogens used in the present invention include phenol, 2-chlorophenol, 4-chlorophenol,
Phenol derivatives such as 2,4-dichlorophenol,
Or aniline, N, N-dimethylaniline, N, N
-Diethylaniline, N, N-diethyl-m-toluidine, N, N-diethyl-m-anisidine, N-ethyl-
N- (3-methylphenyl) -N'-acetylethylenediamine, N-ethyl-N- (2-hydroxy-3-sulfopropyl) -m-anisidine, N-ethyl-N-
Examples thereof include aniline derivatives such as (2-hydroxy-3-sulfopropyl) -m-toluidine, N-ethyl-N-sulfopropyl-m-toluidine and N-ethyl-N-sulfopropyl-m-anisidine.

The ascorbate oxidase used in the present invention is obtained from plants of the Cucurbitaceae family such as pumpkin and cucumber, derived from microorganisms, produced by gene recombination, and these enzymes are chemically modified. You can list things such as things.

Examples of the organic acid manganese salt to be used include manganese acetate, manganese lactate, ethylenediaminetetraacetic acid manganese and the like. Examples of manganese salts of inorganic acids include manganese chloride, manganese bromide, manganese phosphate, manganese borate, manganese carbonate and the like.

The reagent composition for measuring biological components of the present invention comprises an oxidase, a peroxidase, a chromogen, a coupler, an ascorbate oxidase and a manganese salt of an organic acid or an inorganic acid. The composition contains an enzyme that produces a substance that serves as a substrate for an oxidase, depending on the component to be measured. The composition may be in the form of a liquid reagent, a freeze-dried preparation, or a combination of solutions. The liquid reagent contains an appropriate buffer, and may be one or two liquids.

The content of ascorbate oxidase in the reagent composition is not particularly limited, but it is preferably 0.1 to 50 units (U) / ml in the reagent composition.
The content of the manganese salt of an organic acid or an inorganic acid is preferably 0.01 to 1 mM in the reagent composition. When the content of manganese salt exceeds 1 mM, blank color development tends to increase, which causes a problem in measurement precision. If the manganese salt content is less than 0.01 mM, the effect of avoiding the influence of ascorbic acid is not sufficient. When the reagent composition is a two-part reagent, ascorbate oxidase is preferably present in the first reagent, and the manganese salt of an organic acid or an inorganic acid may be present in either the first reagent or the second reagent. The amount of the oxidase, the enzyme that produces a substance that serves as a substrate for the oxidase, the peroxidase, the chromogen, and the coupler is not particularly limited as long as it does not interfere with the measurement.

In the measuring method of the present invention, the above-mentioned reagent composition is allowed to act on a biological sample, and at the same time as or after the interfering substances in the biological components are removed, hydrogen peroxide produced by an oxidase is converted into peroxidase and a color. The biological components in the biological sample are measured by measuring the absorbance of the dye formed by the action of the drug substance and the coupler. Therefore, the operation to act ascorbic acid oxidase and manganese salt of organic acid or inorganic acid to remove interfering substances in biological components is
It may be carried out in any of an operation of causing an oxidase to act on a biological component in a sample or a substance derived from the biological component, and an operation of measuring the produced hydrogen peroxide.

[0023]

According to the present invention, it is possible to avoid the influence of ascorbic acid contained in a biological sample on the measurement of biological components, without pretreatment of the sample. Similar effects cannot be obtained by combining ascorbate oxidase with other metal salts such as copper salts or zinc salts. Also, compared to the addition of ascorbic acid alone,
It is possible to avoid the influence of ascorbic acid up to high concentrations on the measurement of biological components. Further, even when ascorbic acid oxidase containing a stabilizer is used, it is possible to avoid the influence of ascorbic acid on biological components up to high concentrations on the measurement of biological components. Further, the effect of the present invention is to avoid interference with the measured value of ascorbic acid, and does not exert a remarkable effect on the stabilization of ascorbate oxidase.

[0024]

EXAMPLES The present invention will be described in detail below with reference to examples. In the examples, abbreviations indicate the following. TOOS: N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3,5-dimethylaniline ATP: adenosine triphosphate TES: N-tris (hydroxymethyl) methyl-2-
Aminoethanesulfonic acid TAPS: N-tris (hydroxymethyl) methyl-3
-Aminopropanesulfonic acid EDTA / Zn: disodium ethylenediaminetetraacetate / zinc EDTA / Cu: disodium ethylenediaminetetraacetate / copper EDTA / Mn: disodium ethylenediaminetetraacetate / manganese

Example 1 Measurement of Free Cholesterol in Serum The following reagents A to C were prepared. Reagent A: (First reagent) Peroxidase 10u / ml TOOS 2.0mM phosphate buffer 100mM (Second reagent) Cholesterol oxidase 1.5u / ml 4-Aminoantipyrine 0.6mM phosphate buffer 100mM Reagent B: (No. 1 reagent) Peroxidase 10u / ml TOOS 2.0mM phosphate buffer 100mM Ascorbate oxidase 0.2U / ml (from pumpkin) (2nd reagent) Cholesterol oxidase 1.5u / ml 4-aminoantipyrine 0.6mM phosphate buffer Liquid 100 mM Reagent C: (first reagent) Peroxidase 10 u / ml TOOS 2.0 mM Phosphate buffer 100 mM Ascorbate oxidase 0.2 U / ml (from pumpkin) Manganese chloride 0.2 mM (second reagent) Cholesterol Oxidase 1.5 U / ml 4-aminoantipyrine 0.6mM phosphate buffer 100mM

The following samples 1 to 6 were prepared as samples. Sample 1 person pool serum: distilled water = 9: 1 sample 2 person pool serum: 200 mg / dl ascorbic acid aqueous solution = 9: 1 sample 3 person pool serum: 400 mg / dl ascorbic acid aqueous solution = 9: 1 sample 4 person pool serum: 600 mg / dl ascorbic acid aqueous solution = 9: 1 sample 5 person pool serum: 800 mg / dl ascorbic acid aqueous solution = 9: 1 sample 6 person pool serum: 1000 mg / dl ascorbic acid aqueous solution = 9: 1

To 4 μl of each sample, 200 μl of the first reagent of each of the reagents A to C was added, and after heating at 37 ° C. for 5 minutes, the reagent A was added.
After adding 100 μl of the second reagent of -C and heating at 37 ° C. for 5 minutes, the absorbance at 546 nm was measured using purified water as a control. Based on the obtained absorbance, the free cholesterol concentration was determined from a calibration curve prepared in advance using a standard solution. The results are shown in Table 2. In the table, the numbers indicate the unit mg / dl.

[0028]

[Table 2]

As is apparent from Table 1, the reagent A cannot be accurately measured for free cholesterol due to the influence of ascorbic acid. Further, the reagent B that acts only ascorbic acid oxidase is affected by ascorbic acid in a sample having a high ascorbic acid concentration. However, with the reagent C of the present invention, the influence can be avoided even in a sample having a high ascorbic acid concentration.

Example 2 Measurement of Uric Acid in Serum The following reagents D to F were prepared. Reagent D: (1st reagent) Peroxidase 10u / ml TOOS 2.0mM phosphate buffer 100mM (2nd reagent) Uricase 1u / ml 4-aminoantipyrine 0.6mM phosphate buffer 100mM Reagent E: (1st reagent) Peroxidase 10u / ml TOOS 2.0mM phosphate buffer 100mM Ascorbate oxidase 0.2u / ml (from pumpkin) (2nd reagent) Uricase 1u / ml 4-aminoantipyrine 0.6mM phosphate buffer 100mM reagent F: ( 1st reagent) Peroxidase 10u / ml TOOS 2.0mM phosphate buffer 100mM Ascorbate oxidase 0.2U / ml (from pumpkin) Manganese chloride 0.2mM (2nd reagent) Uricase 1u / ml 4-aminoantipyrine 0.6m The phosphate buffer 100mM sample was prepared the same sample 1-6 of Example 1.

To 6 μl of each sample, 200 μl of the first reagent of each of reagents D to F was added, and after heating at 37 ° C. for 5 minutes, 100 μl of the second reagent of each of reagents D to F was added, and then added at 37 ° C. for 5 minutes. After warming, the absorbance at 546 nm was measured using purified water as a control.
Based on the obtained absorbance, the uric acid concentration was determined from a calibration curve prepared in advance using a standard solution. The results are shown in Table 3. In the table, the numbers indicate the unit mg / dl.

[0032]

[Table 3]

As is clear from Table 2, the reagent F of the present invention can avoid the influence of interfering substances even in a sample having a high ascorbic acid concentration.

Example 3 Measurement of Neutral Fat The following reagents G to I were prepared. Reagent G: (First reagent) Glycerol kinase 1 u / ml Glycerol-3-phosphate oxidase 5 u / ml Peroxidase 10 u / ml TOOS 2 mM ATP 3 mM TES buffer 50 mM (Second reagent) Lipoprotein lipase 3 u / ml 4-aminoantipyrine 0.6 mM TES buffer 50 mM Reagent H: (first reagent) Glycerol kinase 1 u / ml Glycerol-3-phosphate oxidase 5 u / ml Peroxidase 10 u / ml TOOS 2 mM ATP 3 mM TES buffer 50 mM Ascorbate oxidase 0.2 U / ml (From pumpkin) (2nd reagent) lipoprotein lipase 3u / ml 4-aminoantipyrine 0.6mM TES buffer 50mM

Reagent I: (First reagent) Glycerol kinase 1 u / ml Glycerol-3-phosphate oxidase 5 u / ml Peroxidase 10 u / ml TOOS 2 mM ATP 3 mM TES buffer 50 mM Ascorbate oxidase 0.2 U / ml (from pumpkin) Manganese chloride 0.2 mM (second reagent) Lipoprotein lipase 3 u / ml 4-aminoantipyrine 0.6 mM TES buffer 50 mM As samples, the same samples 1 to 6 as in Example 1 were prepared.

250 μl of the first reagent of reagents G to I was added to 3 μl of each sample and heated at 37 ° C. for 5 minutes.
After adding 125 μl of the second reagent of I and heating at 37 ° C. for 5 minutes, the absorbance at 546 nm was measured using purified water as a control. Based on the obtained absorbance, the neutral fat concentration was determined from a calibration curve prepared in advance using a standard solution. The results are shown in Table 4. Numerical values in the table indicate the unit mg / dl.

[0037]

[Table 4]

As is clear from Table 3, the reagent I of the present invention can avoid the influence of interfering substances even in a sample having a high ascorbic acid concentration.

Example 4 Measurement of Serum Phospholipid The following reagents J to L were prepared. Reagent J: (First reagent) Phospholipase D 0.5u / ml Peroxidase 10u / ml TOOS 2mM Tris buffer 50mM (Second reagent) Choline oxidase 10u / ml 4-Aminoantipyrine 0.6mM Tris buffer 50mM

Reagent K: (First reagent) Phospholipase D 0.5u / ml Peroxidase 10u / ml TOOS 2mM Tris buffer 50mM Ascorbate oxidase 0.2U / ml (from pumpkin) (Second reagent) Choline oxidase 10u / Ml 4-aminoantipyrine 0.6 mM Tris buffer 50 mM Reagent L: (first reagent) Phospholipase D 0.5 u / ml peroxidase 10 u / ml TOOS 2 mM Tris buffer 50 mM ascorbate oxidase 0.2 U / ml (pumpkin Origin) Manganese chloride 0.2 mM (second reagent) Choline oxidase 10 u / ml 4-aminoantipyrine 0.6 mM Tris buffer 50 mM As samples, the same samples 1 to 6 as in Example 1 were prepared.

250 μl of the first reagent of reagents J to L was added to 3 μl of each sample, and after heating at 37 ° C. for 5 minutes, the reagents J to L
After adding 125 μl of L of the second reagent and heating at 37 ° C. for 5 minutes, the absorbance at 546 nm was measured using purified water as a control. Based on the obtained absorbance, the phospholipid concentration was determined from a calibration curve prepared in advance using a standard solution. The results are shown in Table 5. Numerical values in the table indicate the unit mg / dl.

[0042]

[Table 5]

As is clear from Table 4, the reagent L of the present invention can avoid the influence of interfering substances even in a sample having a high ascorbic acid concentration.

Example 5 Measurement of serum creatinine The following reagents M to O were prepared. Reagent M: (first reagent) creatine amidinohydrolase 100u / ml sarcosine oxidase 10u / ml peroxidase 10u / ml TOOS 2mM TAPS buffer 50mM (second reagent) creatinine amide hydrolase 300u / ml 4-aminoantipyrine 0.6mM TAPS buffer Liquid 50 mM Reagent N: (first reagent) Creatine amidinohydrolase 100 u / ml sarcosine oxidase 10 u / ml peroxidase 10 u / ml TOOS 2 mM TAPS buffer 50 mM ascorbate oxidase 0.2 U / ml (from pumpkin) (second reagent) creatinine Amidohydrolase 300u / ml 4-aminoantipyrine 0.6mM TAPS buffer 50mM

Reagent O: (first reagent) creatine amidinohydrolase 100u / ml sarcosine oxidase 10u / ml peroxidase 10u / ml TOOS 2mM TAPS buffer 50mM ascorbate oxidase 0.2U / ml (from pumpkin) manganese chloride 0.2mM (Second reagent) Creatinine amide hydrolase 300 u / ml 4-aminoantipyrine 0.6 mM TAPS buffer 50 mM As samples, the same samples 1 to 6 as in Example 1 were prepared.

To 6 μl of each sample, 300 μl of the first reagent of reagents M to O was added, and after heating at 37 ° C. for 5 minutes, the reagents M to O were added.
After adding 100 μl of the second reagent of O and heating at 37 ° C. for 5 minutes, the absorbance at 546 nm was measured using purified water as a control. The creatinine concentration was determined from a calibration curve prepared in advance using a standard solution based on the obtained absorbance. The results are shown in Table 6
Shown in. In the table, the numerical values indicate the unit mg / dl.

[0047]

[Table 6]

As is clear from Table 5, the reagent O of the present invention can avoid the influence of interfering substances even in a sample having a high ascorbic acid concentration.

Example 6 Measurement of serum uric acid The following reagents P to S were prepared. Reagent P: (1st reagent) Peroxidase 10u / ml TOOS 2.0mM phosphate buffer 100mM (2nd reagent) Uricase 1u / ml 4-aminoantipyrine 0.6mM phosphate buffer 100mM Reagent Q: (1st reagent) Peroxidase 10u / ml TOOS 2.0mM phosphate buffer 100mM Ascorbate oxidase 0.2U / ml (from pumpkin) EDTA.Zn 0.2mM (second reagent) Uricase 1u / ml 4-aminoantipyrine 0.6mM phosphate buffer Liquid 100 mM

Reagent R: (First reagent) Peroxidase 10 u / ml TOOS 2.0 mM Phosphate buffer 100 mM Ascorbate oxidase 0.2 U / ml (from pumpkin) EDTA / Cu 0.2 mM (second reagent) Uricase 1 u / ml 4-Aminoantipyrine 0.6 mM Phosphate buffer 100 mM Reagent S: (First reagent) Peroxidase 10 u / ml TOOS 2.0 mM Phosphate buffer 100 mM Ascorbate oxidase 0.2 U / ml (from pumpkin) EDTA Mn 0 .2 mM (second reagent) uricase 1 u / ml 4-aminoantipyrine 0.6 mM phosphate buffer 100 mM As samples, the same samples 1 to 6 as in Example 1 were prepared.

To 6 μl of each sample, 200 μl of the first reagent of each of the reagents P to S was added, and after heating at 37 ° C. for 5 minutes, 100 μl of the second reagent of each of the reagents P to S was added, followed by addition of 5 μl at 37 ° C.
After warming for minutes, the absorbance at 546 nm was measured using purified water as a control. Based on the obtained absorbance, the uric acid concentration was determined from a calibration curve prepared in advance using a standard solution. The results are shown in Table 7. In the table, the numbers indicate the unit mg / dl.

[0052]

[Table 7]

As is clear from Table 6, the influence of the reagent S of the present invention containing EDTA.Mn can be avoided even in a sample having a high ascorbic acid concentration.

Example 7 Measurement of Serum Uric Acid The following reagents T to V were prepared. Reagent T: (First reagent) Peroxidase 10u / ml TOOS 2.0mM phosphate buffer 100mM (Second reagent) Uricase 1u / ml 4-aminoantipyrine 0.6mM phosphate buffer 100mM

Reagent U: (First reagent) Peroxidase 10u / ml TOOS 2.0mM phosphate buffer 100mM Ascorbate oxidase 0.5u / ml (from pumpkin) (Second reagent) Uricase 1u / ml 4-aminoantipyrine 0 6 mM phosphate buffer 100 mM reagent V: (first reagent) peroxidase 10 u / ml TOOS 2.0 mM phosphate buffer 100 mM ascorbate oxidase 0.5 u / ml (from pumpkin) manganese chloride 0.2 mM (reagent 2) uricase 1 u / ml 4-aminoantipyrine 0.6 mM phosphate buffer 100 mM As samples, the same samples 1 to 6 as in Example 1 were prepared.

200 μl of the first reagent of each reagent T to V was added to 6 μl of the sample, and after heating at 37 ° C. for 5 minutes, each reagent T
After adding 100 μl of the second reagent of -V and heating at 37 ° C for 5 minutes, the absorbance at 546 nm was measured using purified water as a control. Based on the obtained absorbance, the uric acid concentration was determined from a calibration curve prepared in advance using a standard solution. Each of the reagents U to V prepared in the same manner was stored at 37 ° C. for 3 days and then measured. The results are shown in Table 8. In the table, the numbers indicate the unit mg / dl.

[0057]

[Table 8]

As is clear from Table 7, the effect of the reagent V of the present invention can be avoided even in a sample having a high ascorbic acid concentration. Further, the effect is more remarkable for the reagent stored at 37 ° C. for 3 days.

Claims (14)

[Claims] 1. A method for measuring a biological component by causing an oxidase to act on a biological component or a substance derived from the biological component in a sample and measuring the produced hydrogen peroxide, wherein ascorbic acid oxidase and an organic acid or A method for measuring a biological component, which comprises reacting a manganese salt of an inorganic acid to remove interfering substances in the biological component. 2. The method for measuring a biological component according to claim 1, wherein the produced hydrogen peroxide is quantified using a peroxidase, a chromogen and a coupler. 3. The manganese salt of an organic acid or an inorganic acid is manganese acetate, manganese lactate, ethylenediamine tetraacetate manganese, manganese chloride, manganese bromide, manganese phosphate, manganese borate or manganese carbonate. Item 1. A method for measuring a biological component according to item 1. 4. The method for measuring a biological component according to claim 1, wherein the content of the manganese salt of an organic acid or an inorganic acid is 0.01 to 1 mM in the reagent composition. 5. The biological component is glucose, free cholesterol, ester cholesterol, neutral fat, phospholipid, inorganic phosphorus, creatine, creatinine, free fatty acid, urea nitrogen, pyruvic acid or uric acid. 1. The method for measuring a biological component according to 1. 6. An oxidase, a peroxidase, a chromogen,
A reagent composition for measuring a biological component, which comprises a coupler, ascorbate oxidase, and a manganese salt of an organic acid or an inorganic acid. 7. A biocomponent comprising an enzyme that produces a substrate for an oxidase from the biocomponent, an oxidase, a peroxidase, a chromogen, a coupler, an ascorbate oxidase, and a manganese salt of an organic acid or an inorganic acid. Measuring reagent composition. 8. The manganese salt of an organic acid or an inorganic acid is manganese acetate, manganese lactate, ethylenediaminetetraacetate manganese, manganese chloride, manganese bromide, manganese phosphate, manganese borate or manganese carbonate. Item 6. A reagent composition for measuring a biological component according to item 6 or 7. 9. The reagent composition for measuring a biological component according to claim 6, wherein the content of the manganese salt of an organic acid or an inorganic acid is 0.01 to 1 mM in the reagent composition. object. 10. The reagent composition for measuring a biological component according to claim 6, wherein the biological component is glucose, free cholesterol, pyruvic acid or uric acid. 11. The oxidase is glucose oxidase,
The reagent composition for measuring a biological component according to claim 6, which is cholesterol oxidase, pyruvate oxidase or uricase. 12. The reagent composition for measuring a biological component according to claim 7, wherein the biological component is ester cholesterol, neutral fat, phospholipid, inorganic phosphorus, creatine, creatinine, free fatty acid or urea nitrogen. object. 13. An enzyme that produces a substrate for an oxidase from a biological component is cholesterol esterase, lipoprotein lipase, glycerin kinase, phospholipase D, purine nucleoside phosphorylase, creatine amidinohydrolase, creatinine amide hydrolase, acyl CoA synthetase. Alternatively, the reagent composition for measuring a biological component according to claim 7, which is ureamide lyase. 14. The oxidase is cholesterol oxidase, glycerol oxidase, glycerol-3-phosphate oxidase, choline oxidase, xanthine oxidase, sarcosine oxidase, acyl CoA oxidase or pyruvate oxidase. Reagent composition for measuring biological components of.