CN111579515A - Method for oxidizing TMB by using nano-cerium dioxide as simulated oxidase - Google Patents
Method for oxidizing TMB by using nano-cerium dioxide as simulated oxidase Download PDFInfo
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- 229910000422 cerium(IV) oxide Inorganic materials 0.000 title claims abstract description 36
- 102000004316 Oxidoreductases Human genes 0.000 title claims abstract description 28
- 108090000854 Oxidoreductases Proteins 0.000 title claims abstract description 28
- 238000000034 method Methods 0.000 title claims abstract description 15
- 230000001590 oxidative effect Effects 0.000 title claims abstract description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 24
- 230000003278 mimic effect Effects 0.000 claims abstract description 19
- 230000003197 catalytic effect Effects 0.000 claims abstract description 15
- CETPSERCERDGAM-UHFFFAOYSA-N ceric oxide Chemical compound O=[Ce]=O CETPSERCERDGAM-UHFFFAOYSA-N 0.000 claims abstract description 13
- 230000003647 oxidation Effects 0.000 claims abstract description 13
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 13
- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims abstract description 8
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims abstract description 8
- 239000001267 polyvinylpyrrolidone Substances 0.000 claims abstract description 8
- NHWVNLMOFQZFMZ-UHFFFAOYSA-H cerium(3+) trisulfate tetrahydrate Chemical compound O.O.O.O.[Ce+3].[Ce+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O NHWVNLMOFQZFMZ-UHFFFAOYSA-H 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 28
- MEXSQFDSPVYJOM-UHFFFAOYSA-J cerium(4+);disulfate;tetrahydrate Chemical compound O.O.O.O.[Ce+4].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O MEXSQFDSPVYJOM-UHFFFAOYSA-J 0.000 claims description 4
- 230000008859 change Effects 0.000 claims description 4
- 238000002835 absorbance Methods 0.000 claims description 3
- 239000008055 phosphate buffer solution Substances 0.000 claims description 3
- 229910052684 Cerium Inorganic materials 0.000 claims description 2
- GWXLDORMOJMVQZ-UHFFFAOYSA-N cerium Chemical compound [Ce] GWXLDORMOJMVQZ-UHFFFAOYSA-N 0.000 claims description 2
- 239000003607 modifier Substances 0.000 claims description 2
- 239000012716 precipitator Substances 0.000 claims 1
- 102000004190 Enzymes Human genes 0.000 abstract description 13
- 108090000790 Enzymes Proteins 0.000 abstract description 13
- 230000015572 biosynthetic process Effects 0.000 abstract description 7
- 238000003786 synthesis reaction Methods 0.000 abstract description 7
- 230000004044 response Effects 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 239000003054 catalyst Substances 0.000 abstract description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 2
- 229910052760 oxygen Inorganic materials 0.000 abstract description 2
- 239000001301 oxygen Substances 0.000 abstract description 2
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 abstract 5
- 239000002994 raw material Substances 0.000 abstract 1
- 239000000126 substance Substances 0.000 abstract 1
- 238000006243 chemical reaction Methods 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 5
- 238000003756 stirring Methods 0.000 description 4
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- 238000002441 X-ray diffraction Methods 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000003851 biochemical process Effects 0.000 description 1
- OZECDDHOAMNMQI-UHFFFAOYSA-H cerium(3+);trisulfate Chemical compound [Ce+3].[Ce+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O OZECDDHOAMNMQI-UHFFFAOYSA-H 0.000 description 1
- 229910000333 cerium(III) sulfate Inorganic materials 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000004146 energy storage Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 229910001404 rare earth metal oxide Inorganic materials 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
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- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
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- 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
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- G01N23/207—Diffractometry using detectors, e.g. using a probe in a central position and one or more displaceable detectors in circumferential positions
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Abstract
The invention discloses a method for oxidizing TMB (3, 3',5,5' -tetramethyl benzidine) by using nano-cerium dioxide as a simulated oxidase, which comprises the steps of preparing the TMB at room temperature by using tetrahydrate cerium sulfate, polyvinylpyrrolidone and sodium hydroxide as raw materials; the nano cerium dioxide is used as a catalyst, and is used for directly catalyzing oxygen to oxidize TMB into a blue-green substance within a certain pH range, so that the method is a TMB catalytic oxidation method capable of replacing natural enzymes. The method belongs to the technical field of catalytic oxidation of mimic enzyme, and the nano cerium dioxide mimic oxidase has the advantages of simple synthesis steps, low synthesis cost, rapid pH response, obvious oxidation effect and the like, and has important application prospects in the fields of biological catalytic oxidation, medical immunodetection and the like.
Description
Technical Field
The invention provides a method for catalytic oxidation of TMB by using nano-ceria as a simulated oxidase, belonging to the technical field of catalytic oxidation of the simulated enzyme.
Background
The natural enzyme plays a role in maintaining biochemical processes such as metabolism and genetic information transmission in organisms, and is also a catalyst necessary for catalyzing various chemical reactions in the organisms. Natural enzymes in the body are mostly composed of proteins, and require specific temperature and pH to catalyze chemical reactions in the body. The natural enzyme has high efficiency and specificity, but has the defects of high in vitro production cost, easily influenced property by environmental factors and the like. The artificially synthesized nano mimic enzyme with natural enzyme characteristics has the advantages of stable structure, stable property, simple synthesis process, reusability and the like. The nanometer mimic enzyme can mimic the catalytic activity of natural enzymes, even can mimic two or more natural enzymes, and has the same catalytic mechanism as the natural enzymes.
The rare earth oxides such as nano cerium dioxide and the like are widely applied to the fields of industrial catalysis, energy storage and conversion, biological detection and the like due to low cost, simple preparation and stable physicochemical property. However, there is no report in the prior art about the use of nano-ceria as a mimic oxidase for the catalytic oxidation of TMB. The invention provides a method for catalyzing and oxidizing TMB by using nano cerium dioxide as a simulated oxidase, which has the advantages of rapid pH response, obvious oxidation effect and important application prospect in the fields of biological catalysis, medical detection and the like.
Disclosure of Invention
The invention aims to provide a preparation method of nano cerium dioxide mimic oxidase for catalytic oxidation of TMB, the prepared nano cerium dioxide mimic oxidase is used as a catalyst, can catalyze oxygen in a solution to oxidize TMB within a certain pH range, and has the characteristics of simple synthesis steps, low synthesis cost, rapid pH response, remarkable oxidation effect and the like.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows.
The nano cerium dioxide as simulating oxidase is prepared with cerium sulfate tetrahydrate as cerium source, polyvinyl pyrrolidone as surface modifier and sodium hydroxide as precipitant at room temperature. The method comprises the following specific steps: dissolving polyvinylpyrrolidone in deionized water containing tetrahydrate cerium sulfate, adding sodium hydroxide to form a mixed solution after uniformly stirring, and stirring for 30 minutes at room temperature to obtain the nano cerium dioxide.
In the scheme, the molar ratio of the cerium sulfate tetrahydrate, the sodium hydroxide and the polyvinylpyrrolidone is 1: 3: 9.
A method for catalyzing and oxidizing TMB by using the nano-ceria as a simulated oxidase specifically comprises the following steps:
(1) the TMB solution was dispersed in phosphate buffer at a certain pH to form solution I.
The above scheme (1) is characterized in that the concentration of TMB in the solution I is 2 mg ml-1The pH range is 5.4-7.0.
(2) Adding the nano cerium dioxide mimic oxidase into the solution I to form a solution II, slightly shaking the solution II to fully mix the solution II, and incubating the solution II at room temperature in a dark place for 5 minutes to obtain a solution III.
The above-mentioned means (2) is characterized in that the concentration of the nano-ceria mimic oxidase is 0.2 mg ml-1。
(3) And (3) putting the solution III into an ultraviolet spectrophotometer, and observing the change of the absorbance.
Compared with the prior art, the invention has the remarkable characteristics that:
(1) the nano cerium dioxide mimic oxidase has wide pH application range and catalytic activity of pH response, can catalyze and oxidize TMB within a certain pH range, has rapid color change reaction and high catalytic oxidation efficiency, and can effectively overcome the defect that natural enzyme is easy to inactivate under an acidic condition.
(2) The preparation method of the nano cerium dioxide mimic oxidase has the advantages of simple synthesis steps, easily-reached reaction temperature, short synthesis time and high purity, and can meet the requirements of large-scale production.
Drawings
FIG. 1 is an XRD diffraction pattern of nano-ceria simulated oxidase prepared in example 1;
FIG. 2 is a TEM image of the nano-ceria mimic oxidase prepared in example 1;
FIG. 3 is a UV absorption spectrum of TMB catalyzed by nano-ceria mimic oxidase prepared in example 1 at different pH. The a curve is an ultraviolet absorption curve obtained under the condition of pH = 5.4; the b curve is an ultraviolet absorption curve obtained under the condition of pH = 6.0; the c curve is an ultraviolet absorption curve obtained under the condition of pH = 6.4; the d curve is the ultraviolet absorption curve obtained under the condition of pH =7.0, and the e curve is the ultraviolet absorption curve obtained under the condition of pH = 7.4.
Detailed Description
The present invention will be further illustrated by the following examples.
Example 1
0.22 g of cerium sulfate tetrahydrate and 0.3g of sodium hydroxide were dissolved in 50 ml of deionized water, respectively. And dissolving the polyvinylpyrrolidone in a cerous sulfate tetrahydrate solution, and continuously stirring at room temperature for 30 minutes until the polyvinylpyrrolidone is completely dissolved. Dropwise adding sodium hydroxide solution into the solution, continuously stirring for 30 minutes at room temperature, centrifuging, washing with deionized water and absolute ethyl alcohol for three times respectively, and drying at 30 ℃ to obtain light yellow powder.
The nano-ceria obtained in the above example is used for catalyzing TMB color reaction, and the specific embodiment is as follows: dispersing 16mg of TMB in 7.5 ml of phosphate buffer solution with a series of pH value gradients (pH gradients of 5.4, 6.0, 6.4, 7.0 and 7.4) to form a solution system I; and adding 1.6 mg of the nano-ceria simulated oxidase prepared in the embodiment into the solution II, slightly oscillating, incubating for 5 minutes in a dark place at room temperature to obtain a solution system III, and observing the change of the absorbance of the solution system III by using an ultraviolet spectrophotometer.
FIG. 1 is an X-ray diffraction pattern of the nano-ceria simulated oxidase obtained in the above example, and it can be seen that the obtained product is ceria.
FIG. 2 is a transmission electron microscope image of the nano-ceria simulated oxidase obtained in the above example, wherein it can be seen that the prepared nano-ceria is in a particulate form.
FIG. 3 is a graph of the effect of the nano-ceria mimic oxidase obtained in the above example on TMB catalytic oxidation under a series of pH value gradients, wherein it is shown that nano-ceria cannot catalyze TMB color development when a phosphate buffer solution with pH of 7.4 is used; as the pH value is reduced from 7.0 to 5.4, the catalytic effect of the nano-cerium dioxide simulated oxidase is gradually enhanced, and the catalytic effect with a pH response is shown.
This example is only one of the preferred embodiments of the present invention, and it should be clear to those skilled in the art that the present invention is not limited by the examples, and that some modifications, alterations and adaptations of the present invention should be made within the scope of the claims of the present invention.
Claims (5)
1. The method for oxidizing TMB by using nano-cerium dioxide as simulated oxidase is characterized in that the nano-cerium dioxide is prepared at room temperature by using tetrahydrate cerium sulfate as a cerium source, polyvinylpyrrolidone as a surface modifier and sodium hydroxide as a precipitator.
2. The nano-ceria mimic oxidase of claim 1, wherein the molar ratio of the cerium sulfate tetrahydrate, the sodium hydroxide and the polyvinylpyrrolidone is 1: 3: 9.
3. A method for catalytic oxidation of TMB by using the nano-ceria mimic oxidase of any of claims 1-2, comprising the steps of:
(1) dissolving TMB in phosphate buffer solution with certain pH value to form solution I;
(1) adding the nano cerium dioxide simulated oxidase into the solution I to form a solution II, slightly shaking the solution II, fully mixing the solution II, and incubating the solution II at room temperature in a dark place for 5 minutes to obtain a solution III;
(3) and (3) putting the solution III into an ultraviolet spectrophotometer, and observing the change of the absorbance.
4. The method as claimed in claim 3, wherein the pH of the solution I is in the range of 5.4 to 7.0 and the TMB concentration is 2 mg ml-1。
5. The method as claimed in claim 3, wherein the concentration of the nano-ceria mimic oxidase in the solution system II is 0.2 mg ml-1。
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114534711A (en) * | 2022-04-25 | 2022-05-27 | 中国农业大学 | Heavy metal Al3+Rapid detection kit and application thereof |
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