阅读说明：本技术 钼蓝杂多酸材料在过氧化氢检测中的应用 (Application of molybdenum blue heteropoly acid material in hydrogen peroxide detection ) 是由 周晶 刘瑜鑫 于 2019-11-20 设计创作，主要内容包括：本发明公开了一种检测过氧化氢浓度的方法。本发明利用钼蓝杂多酸材料的颜色信号的变化实现对过氧化氢的快速、灵敏、准确的定量检测。该方法能灵敏、准确地对待测液体中的过氧化氢进行定量分析,提供了一种新的分析测试方法；本发明方法中使用的材料更加简单,所需仪器的价格也更加低廉,能够实现低成本的灵敏、准确的定量分析；本发明分析检测方法可用于食品、药品和活体样品等样品的检测。(The invention discloses a method for detecting the concentration of hydrogen peroxide. The invention realizes the rapid, sensitive and accurate quantitative detection of the hydrogen peroxide by utilizing the change of the color signal of the molybdenum blue heteropoly acid material. The method can sensitively and accurately carry out quantitative analysis on the hydrogen peroxide in the liquid to be detected, and provides a new analysis and test method; the materials used in the method are simpler, the price of the required instrument is lower, and sensitive and accurate quantitative analysis with low cost can be realized; the analysis and detection method can be used for detecting samples such as food, medicines, living samples and the like.)

1. The application of the molybdenum blue heteropoly acid material in detecting the concentration of hydrogen peroxide.

2. The application of the molybdenum blue heteropoly acid material in detecting the concentration of the biological molecules in the biological molecule solution.

3. A method of detecting hydrogen peroxide concentration, comprising:

1) drawing a standard curve: reacting a plurality of groups of known hydrogen peroxide standard solutions with different concentrations with the aqueous solution of the molybdenum blue heteropoly acid material respectively to obtain different mixed solutions, and measuring the light absorption intensity of the mixed solutions respectively by using an ultraviolet visible spectrum; taking the light absorption intensity as a vertical coordinate and the concentration of the hydrogen peroxide standard solution as a horizontal coordinate, and performing linear fitting to obtain a standard curve;

2) determination of the hydrogen peroxide concentration in the hydrogen peroxide solution to be tested: reacting the hydrogen peroxide solution to be detected with the molybdenum blue heteropoly acid material aqueous solution to obtain a mixed solution, measuring the light absorption intensity of the mixed solution under the same wavelength with the hydrogen peroxide standard solution by using an ultraviolet-visible spectrum, and comparing the measured light absorption intensity with the standard curve in the step 1) to obtain the hydrogen peroxide concentration in the solution to be detected.

4. A method for detecting the concentration of biomolecules in a biomolecule solution to be detected, comprising: mixing and incubating the biomolecule solution to be detected and biological enzyme, reacting the mixture with a molybdenum blue heteropoly acid material aqueous solution to obtain a mixed solution, measuring the light absorption intensity of the mixed solution under the same wavelength with the hydrogen peroxide standard solution by utilizing an ultraviolet visible spectrum, comparing the light absorption intensity with the standard curve in the step 1) of claim 3 to obtain the hydrogen peroxide concentration generated by the biomolecule in the solution to be detected under the catalysis of the biological enzyme, and further calculating to obtain the biomolecule concentration in the solution to be detected.

5. The use according to claim 1 or 2 or the method according to claim 3 or 4, characterized in that: the molybdenum blue heteropoly acid material is molybdenum blue heteropoly acid or a composite material of the molybdenum blue heteropoly acid and other materials;

specifically, the molybdenum blue heteropoly acid is selected from at least one of silicon molybdenum blue heteropoly acid, phosphorus molybdenum blue heteropoly acid and vanadium molybdenum blue heteropoly acid;

in the composite material of the molybdenum blue heteropoly acid and other materials, the mass fraction m of the molybdenum blue heteropoly acid is more than 0 and less than 100 percent;

in the composite material of the molybdenum blue heteropoly acid and other materials, the other materials are inorganic materials or organic materials;

the inorganic material is specifically selected from at least one of transition metals, metal sulfides, metal oxides, metal halides, semiconductor materials, and silicates;

the organic material is specifically selected from polymers; the polymer is selected from polydopamine and polyAt least one of 3, 4-ethylenedioxythiophene, polypyrrole, polyethylene and polystyrene; wherein the number average molecular weight of the polydopamine is specifically 10³～10⁶(ii) a The number average molecular weight of the poly 3, 4-ethylenedioxythiophene is specifically 10⁴～10⁶(ii) a The number average molecular weight of the polypyrrole is specifically 10³～10⁶(ii) a The polyethylene has a number average molecular weight of 10³～10⁹(ii) a The polystyrene has a number average molecular weight of 10³～10⁸。

6. The method of claim 3, wherein: in the step 1), the multiple groups are at least 6 groups;

the concentration of the hydrogen peroxide standard solution is 10-500 mu M;

the molar concentration of the molybdenum blue heteropoly acid material aqueous solution is 0.1 mM-10 mM;

the volume ratio of the molybdenum blue heteropoly acid material aqueous solution to the hydrogen peroxide standard solution is (50-2000) mu L: 100 μ L.

7. The method of claim 3 or 5 or 6, wherein: in the step 2), the volume ratio of the hydrogen peroxide solution to be detected to the molybdenum blue heteropoly acid material aqueous solution is synchronous with the volume ratio of the hydrogen peroxide standard solution to the molybdenum blue heteropoly acid material aqueous solution in the step 1).

8. The method according to claim 4 or 5, characterized in that: the volume ratio of the biomolecule solution to be detected to the molybdenum blue heteropoly acid material aqueous solution is the same as the volume ratio of the hydrogen peroxide standard solution to the molybdenum blue heteropoly acid material aqueous solution in the step 1) in the claim 3;

the biomolecule to be detected can generate hydrogen peroxide under the catalysis of a specific biological enzyme;

the biomolecule to be detected is specifically any one of glucose and L-amino acid;

the biological enzyme is specifically any one of glucose oxidase and L-amino acid oxidase.

9. The method according to any one of claims 3-8, wherein: the hydrogen peroxide standard solution, the hydrogen peroxide solution to be detected and the biomolecule solution to be detected are all in corresponding aqueous solution forms;

the reaction temperature of the reaction is 10-40 ℃; the reaction time is 0.5-60 min; specifically, the reaction is carried out for 10min at 25 ℃.

10. Use or method according to any of claims 1-9, wherein: in the detection, ultraviolet-visible spectroscopy is adopted to measure the light absorption intensity;

the light absorption intensity is the light absorption intensity of light with the wavelength of 400-900 nm; specifically, the absorption intensity at 800 nm.

Technical Field

The invention belongs to the field of analysis and detection, particularly relates to a method for detecting hydrogen peroxide concentration, and more particularly relates to application of a molybdenum blue heteropoly acid material in hydrogen peroxide detection.

Background

Hydrogen peroxide is a small molecule with strong oxidizing properties produced by the organism's natural metabolism, and is a well-known cytotoxic substance. An abnormal amount of hydrogen peroxide in a living body may cause damage to single-stranded DNA, denaturation of proteins, reduction in biological activity of enzymes, reduction in fluidity of cell membranes, peroxidation of cytoplasmic matrix, and the like. In addition, hydrogen peroxide is also a metabolite in a variety of biological enzyme catalytic processes, and has important physiological significance. Therefore, the detection of hydrogen peroxide is extremely important for the study of biological toxicity and physiological processes.

Disclosure of Invention

The invention aims to provide an application of a molybdenum blue heteropoly acid material in hydrogen peroxide detection. The invention realizes the rapid, sensitive and accurate quantitative detection of the hydrogen peroxide by utilizing the change of the color signal of the molybdenum blue heteropoly acid material.

The invention claims application of a molybdenum blue heteropoly acid material in detecting hydrogen peroxide concentration.

The invention claims application of a molybdenum blue heteropoly acid material in detecting the concentration of biomolecules in a biomolecule solution.

The invention claims a method for detecting hydrogen peroxide concentration, which comprises the following steps:

1) drawing a standard curve: reacting multiple groups of known hydrogen peroxide standard solutions with different concentrations with the aqueous solution of the molybdenum blue heteropoly acid material respectively to obtain different mixed solutions, and measuring the light absorption intensity (such as A800) of the mixed solutions by utilizing an ultraviolet-visible spectrum; taking the light absorption intensity (such as A800) as a vertical coordinate, taking the concentration of the hydrogen peroxide standard solution as a horizontal coordinate, and performing linear fitting to obtain a standard curve;

2) determination of the hydrogen peroxide concentration in the hydrogen peroxide solution to be tested: and (2) reacting the hydrogen peroxide solution to be detected with the molybdenum blue heteropoly acid material aqueous solution to obtain a mixed solution, measuring the light absorption intensity (such as A800) of the mixed solution under the same wavelength as the hydrogen peroxide standard solution by using an ultraviolet visible spectrum, and comparing the light absorption intensity with the standard curve in the step 1) to obtain the hydrogen peroxide concentration in the solution to be detected.

The method for detecting the concentration of the biomolecules in the biomolecule solution to be detected, which is provided by the invention, comprises the following steps: mixing and incubating the biomolecule solution to be detected and biological enzyme, reacting the mixture with a molybdenum blue heteropoly acid material aqueous solution to obtain a mixed solution, measuring the light absorption intensity of the mixed solution under the same wavelength with the hydrogen peroxide standard solution by utilizing an ultraviolet visible spectrum, comparing the light absorption intensity with the standard curve in the step 1) of the method to obtain the hydrogen peroxide concentration generated by the biomolecule in the solution to be detected under the catalysis of the biological enzyme, and further calculating to obtain the biomolecule concentration in the solution to be detected.

In the application and the method, the molybdenum blue heteropoly acid material is molybdenum blue heteropoly acid or a composite material of the molybdenum blue heteropoly acid and other materials; the molybdenum blue heteropoly acid material can be prepared by various existing published methods;

specifically, the molybdenum blue heteropoly acid is selected from at least one of silicon molybdenum blue heteropoly acid, phosphorus molybdenum blue heteropoly acid and vanadium molybdenum blue heteropoly acid;

in the composite material of the molybdenum blue heteropoly acid and other materials, the mass fraction m of the molybdenum blue heteropoly acid is more than 0 and less than 100 percent;

in the composite material of the molybdenum blue heteropoly acid and other materials, the other materials are inorganic materials or organic materials;

the inorganic material is specifically selected from at least one of transition metals, metal sulfides, metal oxides, metal halides, semiconductor materials, and silicates; in particular mesoporous silicon dioxide or TiO₂；

The organic material is specifically selected from polymers; the polymer is specifically selected from at least one of polydopamine, poly 3, 4-ethylenedioxythiophene, polypyrrole, polyethylene and polystyrene; wherein the number average molecular weight of the polydopamine is specifically 10³～10⁶(ii) a The number average molecular weight of the poly 3, 4-ethylenedioxythiophene is specifically 10⁴～10⁶(ii) a The number average molecular weight of the polypyrrole is specifically 10³～10⁶(ii) a The polyethylene has a number average molecular weight of 10³～10⁹(ii) a The polystyrene has a number average molecular weight of 10³～10⁸。

The composite material of the molybdenum blue heteropoly acid and other materials can be a mesoporous silica-silicomolybdic blue heteropoly acid nano composite material or TiO₂-phosphomolybdic blue heteropoly acid nanocomposites;

in the step 1), the multiple groups are at least 6 groups;

the concentration of the hydrogen peroxide standard solution is 10-500 mu M, and specifically can be 10 mu M, 100 mu M, 200 mu M, 300 mu M, 400 mu M or 500 mu M;

the molar concentration of the molybdenum blue heteropoly acid material aqueous solution is 0.1 mM-10 mM, and specifically can be 0.5 mM;

the volume ratio of the molybdenum blue heteropoly acid material aqueous solution to the hydrogen peroxide standard solution is (50-2000) mu L: 100 μ L, the specific volume ratio may be 1:1, the volume ratio can be (50-2000) mu L: 100 μ L, the specific volume ratio may be 100 μ L: 100 μ L.

In the step 2), the volume ratio of the hydrogen peroxide solution to be detected to the molybdenum blue heteropoly acid material aqueous solution is synchronous with the volume ratio of the hydrogen peroxide standard solution to the molybdenum blue heteropoly acid material aqueous solution in the step 1).

The volume ratio of the biomolecule solution to be detected to the molybdenum blue heteropoly acid material aqueous solution is the same as the volume ratio of the hydrogen peroxide standard solution to the molybdenum blue heteropoly acid material aqueous solution in the step 1);

the biomolecule to be detected can generate hydrogen peroxide under the catalysis of a specific biological enzyme;

the biomolecule to be detected is specifically any one of glucose and L-amino acid;

the biological enzyme is specifically any one of glucose oxidase and L-amino acid oxidase.

The hydrogen peroxide standard solution, the hydrogen peroxide solution to be detected and the biomolecule solution to be detected are all in corresponding aqueous solution forms;

the reaction temperature of the reaction is 10-40 ℃; the reaction time is 0.5-60 min; specifically, the reaction is carried out for 10min at 25 ℃.

In the detection, ultraviolet-visible spectroscopy is adopted to measure the light absorption intensity;

the light absorption intensity is the light absorption intensity of light with the wavelength of 400-900 nm; specifically the absorption intensity at 800nm (e.g.A 800).

The invention realizes sensitive and accurate quantitative detection of hydrogen peroxide by using the change of the light absorption intensity of the molybdenum blue heteropoly acid material under specific wavelength. Specifically, a linear graph (regression coefficient R) of the value and the concentration of the absorption intensity of a series of solutions of the components to be detected with known concentrations under a specific wavelength is measured²Not less than 0.99), obtaining a standard linear map; and measuring the light absorption intensity of the component to be measured with unknown concentration under the specific wavelength, and comparing with the standard linear map to obtain the target product.

Compared with the prior art, the invention has the following beneficial effects:

1) the method can sensitively and accurately carry out quantitative analysis on the hydrogen peroxide in the liquid to be detected, and provides a new analysis and test method;

2) the materials used in the method are simpler, the price of the required instrument is lower, and sensitive and accurate quantitative analysis with low cost can be realized;

3) the analysis and detection method can be used for detecting samples such as food, medicines, living samples and the like.

Drawings

FIG. 1 is a graph showing the UV-Vis spectra of phosphomolybdic blue heteropoly acid before and after adding hydrogen peroxide solution in example 1 of the present invention.

FIG. 2 is a linear spectrum of phosphomolybdic blue heteropoly acid in example 1 of the present invention for detecting hydrogen peroxide.

FIG. 3 is the UV-Vis spectra of mesoporous silica-SiMoMoN blue heteropoly acid nanocomposites before and after addition of hydrogen peroxide solution in example 2 of the present invention.

FIG. 4 is a linear spectrum of mesoporous silica-silicomolybdic blue heteropoly acid nanocomposite for detecting hydrogen peroxide in example 2 of the present invention.

FIG. 5 shows TiO in example 3 of the present invention₂-ultraviolet-visible spectrum before and after the phosphomolybdic blue heteropoly acid nano composite material is added into the hydrogen peroxide solution.

FIG. 6 shows TiO in example 3 of the present invention₂-detecting the linear spectrum of hydrogen peroxide by using the phosphomolybdic blue heteropoly acid nanocomposite.

FIG. 7 shows TiO in example 3 of the present invention₂Ultraviolet-visible spectrum before and after the phosphomolybdic blue heteropoly acid nano composite material is added into the L-leucine solution treated by the L-amino acid oxidase.

Detailed Description

The present invention will be further illustrated with reference to the following specific examples, but the present invention is not limited to the following examples. The method is a conventional method unless otherwise specified. The starting materials are commercially available from the open literature unless otherwise specified.

The phosphomolybdic blue heteropoly acid used in the following example 1 was prepared as follows:

first, 0.1g of sodium dihydrogen phosphate powder was ultrasonically dispersed in 5mL of deionized water, and then added to a 100mL beaker. 10mL of ammonium molybdate solution (1M) and 5mL of ascorbic acid solution (5M) were added, respectively. Stirring the mixture at room temperature for 30min, distilling the mixture under reduced pressure, and continuously washing the mixture for several times by using 2mL of mixed solution of ethanol and acetone (the volume ratio is 1:1) to obtain the phosphomolybdic blue heteropoly acid.

The mesoporous silica-silicomolybdic blue heteropoly acid nanocomposite used in the following example 2 is prepared by the following method:

first, 0.1g of bromohexadecyltrimethylamine powder was ultrasonically dispersed in 20mL of deionized water, and then added to a 100mL round bottom flask and heated to 70 ℃. 0.04g of solid sodium hydroxide powder and 0.5mL of tetraethyl orthosilicate liquid were added, respectively. After stirring at room temperature for 30s, 1mL of ethyl acetate liquid was added. After stirring for 3 hours, the mixture was centrifuged, and washed with 5mL of ethanol several times to obtain mesoporous silica. 0.1g of the obtained mesoporous silica was dispersed in 5mL of deionized water, and 10mL of ammonium molybdate solution (1M) and 5mL of ascorbic acid solution (5M) were added. Stirring for 60min at 80 ℃, performing centrifugal separation, and washing with deionized water for several times to obtain the mesoporous silica-silicomolybdyl blue heteropoly acid nanocomposite.

TiO used in example 3 described below₂The-phosphomolybdic blue heteropoly acid nano composite material is prepared by the following method:

first, 0.05g of titanium sulfate powder was dissolved in 20mL of oleic acid, and then added to a 100mL three-necked round-bottomed flask, the solution was heated to 300 ℃ under nitrogen protection to react for 1 hour, centrifuged, and washed with 5mL of ethanol several times to obtain TiO₂And (3) nanoparticles. 0.05g of the obtained mesoporous silica was dispersed in 5mL of deionized water, and 1mL of dipalmitoylphosphatidylcholine solution (50mg mL) was added^-1). After 10min of sonication, 10mL of ammonium molybdate solution (1M) and 5mL of ascorbic acid solution (5M) were added. Stirring at room temperature for 30min, centrifuging, washing with deionized water for several times to obtain TiO₂-phosphomolybdic blue heteropoly acid nanocomposites.