CN103977788B - A kind of Cs xwO 3the preparations and applicatio of class Mimetic Peroxidase - Google Patents
A kind of Cs xwO 3the preparations and applicatio of class Mimetic Peroxidase Download PDFInfo
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- CN103977788B CN103977788B CN201410220658.3A CN201410220658A CN103977788B CN 103977788 B CN103977788 B CN 103977788B CN 201410220658 A CN201410220658 A CN 201410220658A CN 103977788 B CN103977788 B CN 103977788B
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- peroxidase
- glucose
- mimetic peroxidase
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Abstract
The invention provides a kind of Cs
xwO
3class Mimetic Peroxidase and its preparation method and application.This Cs
xwO
3class Mimetic Peroxidase has the activity of class peroxidase, and it can catalysis H
2o
2produce a large amount of hydroxyl radical free radicals, efficient oxidation substrate 3,3 ˊ, 5,5 ˊ-tetramethyl benzidine produces chromogenic reaction.With glucose oxidase coupling, by measuring 3,3 ˊ after oxidation, the ultraviolet absorption value of 5,5 ˊ-tetramethyl benzidine, indirect detection goes out glucose in solutions content, detects and is limited to 0.1 μM.Cs
xwO
3the advantages such as nano material is compared to horseradish peroxidase (HRP), and have preparation technology simple, cost is low, and chemism is stable, and its catalytic effect and applicability are all higher than natural peroxide enzyme, can as the substitute of natural peroxide enzyme.
Description
Technical field
The present invention relates to field of nanometer technology and enzyme catalysis field, be specifically related to a kind of Cs
xwO
3class Mimetic Peroxidase and its preparation method and application.
Background technology
Diabetes have now become a kind of disease affecting human normal life, and the concentration of the glucose generally in human body is 3-8mM, and the concentration of glucose in diabetic's body is 9-40mM.Concentration of glucose is a critical index of diagnosing diabetes, having had a lot of detection modes at present for detecting the concentration of glucose sugar, common are: high performance liquid chromatography, electrochemistry, colorimetric method etc.Wherein colorimetric method because having higher sensitivity compared with other method, easy and simple to handle, low cost and other advantages is subject to people and more pays close attention to.And a large amount of glucose sensors is set up based on HRPO colorimetric method, but, native enzyme limited source, purification difficult, expensive, and in order to keep its clean property, all comparatively harsh to the requirement of experiment condition and operating environment, make it apply to be greatly limited, therefore the exploitation of analogue enztme and application study more and more receive the concern of people.
Since Fe
3o
4nano particle be found to have with catalytic property like Horseradish peroxidase after, people investigated a series of nano material with class peroxidase function.The advantages such as this kind of nanometer analogue enztme has that preparation process is easy, storage requirement is simple, cost is low, high temperature resistant and acid and alkali-resistance, become the object of people's primary study, therefore continue to find the new nano material containing catalytic activity, excavate the important directions that novel nanometer analogue enztme is research at present.Many nano material such as Fe with photo-thermal effect are found by our investigation in early stage
3o
4, CuS, Prussian blue, Au etc. have the character of nanometer analogue enztme.On Nanoscale, Cs is reported according to the people such as ChongshenGuo before
xwO
3as optothermal material, at near-infrared place, there is very high absorption, and there is high-conversion rate, therefore may be used for the photo-thermal therapy of tumour.Based on above research, we think Cs
xwO
3nanometer rods also may have catalytic activity and the application of similar analogue enztme, and this imagination is suggested first and verifies.
Summary of the invention
The object of the invention is to be to provide and a kind of Cs
xwO
3class Mimetic Peroxidase and its preparation method and application.Cs
xwO
3nano material can catalysis H
2o
2produce the hydroxyl radical free radical of strong oxidizing property, efficient oxidation 3,3', 5,5'-tetramethyl benzidine (TMB) produces chromogenic reaction.Have that the reaction time is short, colour developing is fast, feature that catalytic effect and applicability are all higher than natural peroxide enzyme, can be applicable to the detection of blood, Determination of Glucose in Food content.
To achieve these goals, the present invention is by the following technical solutions:
A kind of Cs
xwO
3class Mimetic Peroxidase, described Cs
xwO
3class Mimetic Peroxidase is Cs
xwO
3nano material, it has the activity of class peroxidase.
Described Cs
xwO
3nano material energy catalysis H
2o
2produce a large amount of hydroxyl radical free radicals.
Described Cs
xwO
3the application of class Mimetic Peroxidase: with glucose oxidase coupling, for the mensuration of glucose in solutions content.
Cs
xwO
3nano material energy catalysis H
2o
2produce the hydroxyl radical free radical of strong oxidizing property, efficient oxidation substrate 3,3', 5,5'-tetramethyl benzidine produces chromogenic reaction, then by measuring 3,3' after oxidation, the ultraviolet absorption value of 5,5'-tetramethyl benzidine (oxTMB), indirect detection goes out glucose in solutions content.
The range of linearity of glucose detection is: 0.005-1mM, detects and is limited to 0.1 μM.
Described chromogenic reaction is by 3,3', 5,5'-tetramethyl benzidine, Cs
xwO
3nano material and H
2o
2join in cushioning liquid and react, solution is blue by colourless change.
Described chromogenic reaction condition is PBS buffer solution, pH=4.0, and temperature is 35 DEG C, and the concentration of 3,3', 5,5'-tetramethyl benzidine is 0.6mM, H
2o
2concentration be 50mM.
Described Cs
xwO
3the preparation method of class Mimetic Peroxidase: by WCl
6under agitation be dissolved in absolute ethyl alcohol, add CsOHH
2o, after being uniformly dispersed, then adds anhydrous glacial acetic acid; Gained mixed solution is transferred in polytetrafluoroethylene (PTFE) reactor, 235 DEG C of reaction 20h; Centrifugal washing, for several times, then at 60 DEG C, vacuum drying 12h obtains Cs to ethanol purge
xwO
3nano material.
More specifically: 0.2976gWCl
6be dissolved in 40mL absolute ethyl alcohol under vigorous stirring, then by 0.0636gCsOHH
2the yellowish WCl dissolved before O joins
6in ethanolic solution; After mixed solution is uniformly dispersed, the anhydrous glacial acetic acid of 10mL is joined in above-mentioned solution, mixed solution is transferred in the polytetrafluoroethylene (PTFE) reactor of 100mL, at 235 DEG C of reaction 20h, obtain navy blue product, centrifugal washing, for several times, the product finally obtained is vacuum drying 12h at 60 DEG C for ethanol purge.
Cs
xwO
3nano material has the research of class peroxidase activity:
1) Cs
xwO
3nano material can catalysis H
2o
2oxidation substrates produces chromogenic reaction: by Cs
xwO
3nano material, TMB and H
2o
2join 25mM, in the PBS cushioning liquid of pH=4.0, shake up, reaction 30min, then measures the absorption value of 652nm wavelength with ultraviolet specrophotometer.As shown in Figure 1, Cs
xwO
3+ H
2o
2+ TMB experimental group has higher absorption value compared to other control groups, and Cs is described
xwO
3nano material really can catalysis H
2o
2tMB is oxidized to oxTMB.
2) Cs
xwO
3nano material energy catalysis H
2o
2produce hydroxyl radical free radical, the hydroxyl radical free radical produced and substrate terephthaldehyde acid reaction, produce fluorescent substance, its maximum absorption wavelength is 425nm, and along with the carrying out of reaction, fluorescent value is more and more higher, as shown in Figure 2, Cs is described
xwO
3the enzymatic mechanism of nano material is Fenton's reaction mechanism, catalysis H
2o
2produce hydroxyl radical free radical.
Cs
xwO
3nano material is as the application of class peroxidase:
In the aqueous solution, glucose reacts quantitatively can produce H with oxygen under the catalysis of glucose oxidase
2o
2, and Cs
xwO
3nano material can catalysis H
2o
2produce the hydroxyl radical free radical of strong oxidizing property, then TMB is oxidized to oxTMB, by measuring the ultraviolet absorption value of oxTMB, indirect detection goes out glucose in solutions content.
The present invention has investigated the variable of Different factor to impact of the present invention on the basis of technique scheme:
1, cushioning liquid pH used has been investigated to Cs
xwO
3nano material catalysis H
2o
2the impact of oxidation TMB, determines Optimal pH.
2, temperature has been investigated to Cs
xwO
3nano material catalysis H
2o
2the impact of oxidation TMB, determines best temperature.
Beneficial effect of the present invention is:
Late Cambrian Cs of the present invention
xwO
3nano material has the catalytic activity of class peroxidase, and is applied to the mensuration of glucose in solutions content.Cs
xwO
3nano material is used for the mensuration of glucose as class peroxidase, has that the reaction time is short, develop the color the advantage such as fast, and its catalytic effect and applicability are all higher than natural peroxide enzyme.And compared to horseradish peroxidase (HRP), the advantages such as have preparation technology simply, cost is low, and chemism is stable, can as the substitute of natural peroxide enzyme.With glucose oxidase coupling, can build simple, fast, the colorimetric method of low cost for detecting concentration of glucose, detect and be limited to 0.1 μM.
Accompanying drawing explanation
Fig. 1 is Cs
xwO
3the ultraviolet-visible light spectrogram of the chromogenic reaction of nano material catalysis TMB; Ordinate is absorption value, and abscissa is wavelength (nanometer);
Fig. 2 is Cs
xwO
3the research of nano material nano particle catalytic mechanism; Ordinate is fluorescence intensity, and abscissa is for being wavelength (nanometer);
Fig. 3 is Cs
xwO
3nano material TEM figure, the nano material of synthesized nanometer rods pattern, particle be about as 150nm, wide is 15nm;
Fig. 4 is that different pH value is to Cs
xwO
3nano material catalysis H
2o
2the impact of oxidation TMB; Ordinate is relative intensity, and abscissa is pH value;
Fig. 5 is that condition of different temperatures is to Cs
xwO
3nano material catalysis H
2o
2the impact of oxidation TMB; Ordinate is relative intensity, and abscissa is temperature (DEG C);
Fig. 6 is for utilizing Cs
xwO
3the ultraviolet-visible light spectrogram of nanometer rods indirect detection concentration of glucose, ordinate is absorption value, and abscissa is concentration of glucose;
Measure the linear regression collection of illustrative plates of glucose content when Fig. 7 is low concentration, ordinate is absorption value, and abscissa is concentration of glucose.
Detailed description of the invention
embodiment 1
Under temperature 35 DEG C of conditions, by the Cs of 50 μ g/ml
xwO
3nano material joins 2.275ml and is dissolved with TMB(0.6mM) PBS(pH=4.0) in cushioning liquid, add the H of 50mM
2o
2solution, reaction 30min, then measures the absorption value (see figure 1) of 652nm on ultraviolet specrophotometer; Fig. 1 result display Cs
xwO
3group has absorption clearly at 652nm place, and Cs is described
xwO
3nano material has class peroxidase activity, can catalysis H
2o
2oxidation TMB, produces chromogenic reaction.
embodiment 2
At room temperature, by the Cs of 50 μ g/ml
xwO
3nano material adds in the PBS cushioning liquid (pH=4.0) containing the terephthalic acid (TPA) of 1mM, adds 50mMH
2o
2solution, starts timing, now Cs
xwO
3nano material energy catalysis H
2o
2produce hydroxyl radical free radical, the hydroxyl radical free radical produced and substrate terephthaldehyde acid reaction, produce fluorescent substance, its maximum absorption wavelength is 425nm, as shown in Figure 2; Fig. 2 result shows, and along with the reaction time increases, fluorescent value is more and more higher, and illustrate that a large amount of hydroxyl radical free radicals produces, this just demonstrates Cs
xwO
3the catalyst mechanism of nano material is catalysis H
2o
2produce hydroxyl radical free radical.
embodiment 3
By Cs
xwO
3the nano material aqueous solution carries out ultrasonic process 0.5h, then drops on copper mesh, carries out TEM and scan (see figure 3) after drying; Fig. 3 shows gained Cs
xwO
3for nanometer rods pattern, size is more homogeneous, and nanometer rods is about as 150nm, and wide is 15nm.
embodiment 4
At room temperature, by the Cs of 50 μ g/ml
xwO
3the TMB solution of nano material and 0.6mM joins (being pH=2.0 respectively, 3.0,4.0,5.0,6.0,7.0,8.0,9.0,10.0) in the PBS cushioning liquid of the different pH value of 2.275ml, adds the H of 50mM
2o
2solution, start timing, room temperature reaction 30min, then measures the absorption value (see figure 4) of 652nm, the intensity 100% of ultraviolet absorption value corresponding under regulation Optimal pH.
Fig. 4 result shows, and when other condition is constant, sour environment on the weak side is conducive to Cs
xwO
3nano material catalysis H
2o
2the colour developing of oxidation TMB, Optimal pH=4.0.
embodiment 5
By the Cs of 50 μ g/ml
xwO
3the TMB solution of nano material and 0.6mM joins in the pH=4.0PBS cushioning liquid of 2.275ml, and temperature is set to 25 respectively, and 30,35,40,45,50,55,60 DEG C, when adding the H of 50mM
2o
2time, start timing, reaction 30min, then measures the absorption value (see figure 5) of 652nm, and the intensity of ultraviolet absorption value corresponding under regulation optimum temperature is 100%.
Fig. 5 result shows, and when other condition is constant, when temperature is 35 DEG C, TMB colour developing is the most obvious, and Cs is described
xwO
3the optimum temperature that nano material catalysis TMB reacts is 35 DEG C.
embodiment 6
The glucose oxidase being 20mg/mL by 20 μ L concentration adds 600 μ L and is dissolved with in the PBS solution (pH=7.0) of D-Glucose, under temperature 37 DEG C of conditions, react 1h.Then in mixed liquor, add 1.655mLPBS solution (pH=4.0), 150 μ gCs successively
xwO
3be the TMB of 0.6mM with 575 μ L concentration, after mixing, under 35 DEG C of water bath condition, react 1h, survey its UV absorption wavelength (see Fig. 6, Fig. 7) at 652nm place; Fig. 7 result shows, when glucose is in low concentration (0.005-1mM) scope, gained absorption value and concentration of glucose linear, detect be limited to 0.1 μM.
The foregoing is only preferred embodiment of the present invention, all equalizations done according to the present patent application the scope of the claims change and modify, and all should belong to covering scope of the present invention.
Claims (5)
1. a Cs
xwO
3the application of class Mimetic Peroxidase, is characterized in that: with glucose oxidase coupling, for the mensuration of glucose in solutions content; Described Cs
xwO
3class Mimetic Peroxidase is Cs
xwO
3nano material, it has the activity of class peroxidase.
2. Cs according to claim 1
xwO
3the application of class Mimetic Peroxidase, is characterized in that: Cs
xwO
3class Mimetic Peroxidase energy catalysis H
2o
2produce the hydroxyl radical free radical of strong oxidizing property, efficient oxidation substrate 3,3', 5,5'-tetramethyl benzidine produces chromogenic reaction, and then by measuring the ultraviolet absorption value of 3,3', 5,5'-tetramethyl benzidines after oxidation, indirect detection goes out glucose in solutions content.
3. Cs according to claim 1
xwO
3the application of class Mimetic Peroxidase, is characterized in that: the range of linearity of glucose detection is: 0.005-1mM, detects and is limited to 0.1 μM.
4. Cs according to claim 2
xwO
3the application of class Mimetic Peroxidase, is characterized in that: described chromogenic reaction is by 3,3', 5,5'-tetramethyl benzidines, Cs
xwO
3class Mimetic Peroxidase and H
2o
2join in cushioning liquid and react, solution is blue by colourless change.
5. Cs according to claim 2
xwO
3the application of class Mimetic Peroxidase, is characterized in that: described chromogenic reaction condition is PBS buffer solution, pH=4.0, and temperature is 35 DEG C, and the concentration of 3,3', 5,5'-tetramethyl benzidine is 0.6mM, H
2o
2concentration be 50mM.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104596956A (en) * | 2015-01-05 | 2015-05-06 | 上海纳米技术及应用国家工程研究中心有限公司 | Application of nano-nickel oxide as mimetic peroxidase for detecting hydrogen peroxide |
| CN105214646B (en) * | 2015-09-24 | 2018-04-17 | 福建医科大学 | A kind of application of tungsten oxide quanta point material in Mimetic enzyme |
| CN106596409B (en) * | 2015-10-19 | 2019-08-09 | 首都师范大学 | A kind of method of staircase test hydrogenperoxide steam generator concentration |
| CN107561070B (en) * | 2017-08-28 | 2018-11-13 | 菏泽学院 | A kind of glucose quick visualization detection method and its preparation and application |
| CN108976412A (en) * | 2018-06-01 | 2018-12-11 | 南京理工大学 | Application of the Ferrocene and its derivative as Mimetic enzyme of peroxidase |
| CN109116029A (en) * | 2018-07-21 | 2019-01-01 | 福建医科大学 | A method of paper substrate micro-fluidic chip is made based on inkjet printing technology controllability and is detected for glucose quantitation |
| CN108998022B (en) * | 2018-08-29 | 2020-11-24 | 江苏科技大学 | Antimony tin oxide nano-particles with peroxidase-like characteristics and preparation method and application thereof |
| CN110609032A (en) * | 2019-09-03 | 2019-12-24 | 东华理工大学 | Glucose detection method based on application of molybdenum oxide quantum dots as peroxide mimic enzyme |
| CN111965136B (en) * | 2020-08-14 | 2023-09-19 | 陕西科技大学 | Preparation method of peroxide-like nano enzyme beta-FeOOH and application of peroxide-like nano enzyme beta-FeOOH in H 2 O 2 Application in detection |
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| CN102320662A (en) * | 2011-07-04 | 2012-01-18 | 大连工业大学 | Cesium tungsten bronze powder and preparation method thereof |
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| CN102320662A (en) * | 2011-07-04 | 2012-01-18 | 大连工业大学 | Cesium tungsten bronze powder and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| Solvothermal synthesis of caesium tungsten bronze in the presence of various organic acids and its NIR absorption properties;Chongshen Guo, et al;《Materials Science and Engineering》;20111231;第18卷(第3期);摘要、第2页第1段及表1 * |
| Supercritical solvothermal synthesis and neariinfrared absorbing properties of CsxWO3;Chongshen Guo, et al;《Functional Materials Letters》;20120427;第5卷(第2期);全文 * |
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