CN102507691A - Preparation method of ascorbic acid oxidase electrochemical biosensing composite modified electrode - Google Patents
Preparation method of ascorbic acid oxidase electrochemical biosensing composite modified electrode Download PDFInfo
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- CN102507691A CN102507691A CN2011103201600A CN201110320160A CN102507691A CN 102507691 A CN102507691 A CN 102507691A CN 2011103201600 A CN2011103201600 A CN 2011103201600A CN 201110320160 A CN201110320160 A CN 201110320160A CN 102507691 A CN102507691 A CN 102507691A
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Abstract
The invention relates to a preparation method of an ascorbic acid oxidase electrochemical biosensing composite modified electrode. The preparation method comprises the steps of: preparing an electrolyte solution from 3,4-ethylenedioxy thiophene, sodium lauroylsarcosine, carbon nanotubes and 1-ethyl-3-methylimidazolium ethyl sulfate, and carrying out 1.1V constant-potential polymerization in a three-electrode system for 60 seconds to prepare the conducting poly(3,4-ethylenedioxy thiophene) nano film composite modified electrode by one-step reaction; and then drip-coating ascorbic acid oxidase buffer on the surface of the composite modified electrode, and finally covering the enzyme layer with a mixed liquid of carbon nanotubes, Nafion and ionic liquid. The constructed composite modified electrode has the advantages of good biological electric catalysis activity to vitamin C, wide linear range, low detection potential, rapid current response, significant sensitivity, high repeatability and stability, good selectivity, high immobilized enzyme activity and strong affinity; and the preparation method has the characteristics of low cost, simple process, and simplicity and easiness in operation and the like.
Description
Technical field
The invention belongs to the biosensor technology field, relate to a kind of preparation method of ascorbic acid oxidase electrochemica biological sensing composite modified electrode.
Background technology
Since Foulds in 1986 and Lowe glucose oxidase is fixed on make up first conducting polymer glucose oxidase electrochemica biological sensor in the polypyrrole since, conducting polymer as the fixation support of enzyme because advantages such as the premium properties of himself, easy preparation method and simple operations flow process enjoy the extensive favor of biology sensor researcher.The bioactive molecule fixation support that conducting polymer is used for biology sensor has six big advantages: 1. have electron transport ability fast; 2. have the bio-compatibility good with bioactive molecule; 3. can keep the good biologically active of bioactive molecule; 4. can prevent effectively that bioactive molecule runs off; 5. can prolong the life-span of biology sensor; 6. can reduce the cost of biology sensor and be prone to processing.
The conducting polymer enzyme electrode of tradition electrochemical production (enzyme is fixed on the bioelectrode on electrochemical electrode surface by conducting polymer film) bio-compatibility is poor; And enzyme also had certain toxicity (M.Gerard, A.Chaubey, and B. D. Malhotra; Biosens. Bioelectron.; 2002,17,345).Problems such as the result causes poor stability in the testing process, and poor reproducibility, sensitivity are low, detects limit for height, and sensing range is little, and degree of accuracy also has very big influence in practical application.
Summary of the invention
The purpose of this invention is to provide a kind of simple in structure, highly sensitive ascorbic acid oxidase electrochemica biological sensing composite modified electrode and preparation method thereof.
The preparation method of ascorbic acid oxidase electrochemica biological sensing composite modified electrode of the present invention; Be with 3; 4-enedioxy thiophene, carbon nanometer tube, 1-ethyl-3-methylimidazole ethyl-sulfate, sarcosyl are raw material, and constant potential 1.1 V polymerizations preparation in 60 seconds high-performance conductive gathers (3,4-enedioxy thiophene) nano thin-film composite modified electrode in conventional three-electrode system; After the pre-service drying ascorbic acid oxidase is cushioned drop and be coated in the composite modified electrode surface; After the air dry, carbon nanometer tube-Nafion-hydrophobic nonionic liquid mixed liquor is covered the high-performance conductive that contains the ascorbic acid oxidase layer gather (3,4-enedioxy thiophene) nano thin-film composite modified electrode surface; After the air dry, make high-performance ascorbic acid oxidase electrochemica biological sensing composite modified electrode.
The present invention passes through electrochemical method with 3; 4-enedioxy thiophene and carbon nanometer tube (nano material), sarcosyl (bio-compatibility amino acid surfactant), 1-ethyl-3-methylimidazole ethyl-sulfate (hydrophilic " green " electrolyte solution) step make high-performance conductive and gather (3,4-enedioxy thiophene) nano thin-film composite modified electrode.This electrode has combined to gather (3; 4-enedioxy thiophene), the advantage of carbon sodium rice material, bio-compatibility amino acid surfactant, hydrophilic ionic liquid; Not only strengthened and gathered (3; The sensitivity and the affinity of 4-enedioxy thiophene) bio-compatibility, electric conductivity and bio-sensing electrode, but also reduced detectability and detected current potential, impel electron transfer and quicken current-responsive.Method through simple composite drying makes the compound adventitia of bio-compatibility with CNT dispersion liquid, Nafion, one step of hydrophobic nonionic liquid mixing again, has strengthened anti-interference, biological electro catalysis activity, bio-compatibility, stability and the sensitivity of enzyme sensor.
Use the electrochemica biological sensing electrode of this method preparation not only can carry out the detection of success to Vitamin C content in crops and the agricultural product, but also have detect effective, the response time is short, operating potential is low, selectivity is strong, highly sensitive, characteristics such as detectability is low, the range of linearity is wide, affinity is strong, reappearance and good stability, life-span are grown.
Description of drawings
Fig. 1 for the biology sensor of the present invention preparation to ascorbic biological electro catalysis oxidation and voltammetric measuring;
Fig. 2 measures ascorbic biological electro catalysis oxidation and ampere for the biology sensor of the present invention's preparation;
Fig. 3 is the stability of the biology sensor of the present invention's preparation.
Embodiment
A kind of preparation method of ascorbic acid oxidase electrochemica biological sensing composite modified electrode may further comprise the steps:
1, the preparation of electrolyte solution: earlier with 1.42 mg 3; 4-enedioxy thiophene, 2.93 mg sarcosyls, 1 mg percent by weight, 0.1 wt% carbon nanometer tube dispersion liquid, 2.36 mg 1-ethyls-3-methylimidazole ethyl-sulfate ultrasonic dissolution mix at last and obtain electrolyte solution in the deionized water of 1 milliliter of cumulative volume.
2, high-performance conductive gathers (3; 4-enedioxy thiophene) preparation of nano thin-film composite modified electrode: in above-mentioned electrolyte solution, fed nitrogen 20 minutes,, adopt conventional three-electrode system to remove the dissolved oxygen DO in the solution; 1.1 V constant potential polymerizations 60 seconds; One step made gathers (3,4-enedioxy thiophene) nano thin-film composite modified electrode, and whole process is carried out under nitrogen protection.
3, gather (3; 4-enedioxy thiophene) pre-service of nano thin-film composite modified electrode: after the composite modified electrode preparation finishes; Electrode is taken out from electrolyte solution, use deionized water rinsing, to remove the lip-deep 1-ethyl of film composite modified electrode-3-methylimidazole ethyl-sulfate, 3; 4-enedioxy thiophene monomer and oligomer, carbon nanometer tube, sarcosyl, drying for standby then.
4, with 0.3g L
-1Ascorbic acid oxidase damping fluid 5 μ L drip to be coated in and gather (3,4-enedioxy thiophene) nano thin-film composite modified electrode surface, and air dry is subsequent use.
5, the preparation of carbon nanometer tube-Nafion-hydrophobic nonionic liquid complex liquid: earlier with 4% carbon nanometer tube dispersion liquid, 5% Nafion solution, 1-butyl-3-methylimidazole hexafluorophosphoric acid hydrophobic nonionic liquid by volume 1:1 the three is mixed, ultrasonic preparation obtains 0.6 milliliter of carbon nanometer tube of cumulative volume-Nafion-hydrophobic nonionic liquid complex liquid.
6, with carbon nanometer tube-Nafion-hydrophobic nonionic liquid complex liquid cover contain enzyme gather (3,4-enedioxy thiophene) nano thin-film composite modified electrode surface, air dry obtains high-performance ascorbic acid oxidase electrochemica biological sensing composite modified electrode.
7, the enzyme electrode that makes is put into 5 * 10
-2Mol L
– 1Contain in the phosphate buffer (pH 6.5) of sarcosyl, 4 ℃ of preservations are subsequent use.
The parameter optimization of high-performance composite modified electrode and Performance Evaluation: various concentration of material parameters and operating potential in testing process, film forming thickness, temperature, pH value, the phosphate buffer density of electrochemica biological sensor in the preparation process is optimized, and also dynamics, linear range, reappearance, sensitivity, operation and storage stability, the selectivity of enzyme assessed simultaneously.Enzyme dynamics shows that this bio-sensing electrode has good biological affinity and enzymatic activity (apparent Michaelis constant 2.24 mM, apparent maximum current density 1.045 mA cm
-2).And low apparent activation energy value (7.049 kJ M
-1) confirm that further this biology sensor has good biological affinity and enzymatic activity.The bio-sensing combination electrode shows that to ascorbic good biological catalytic oxidation activity this bio-sensing electrode can successfully detect unknown Vitamin C content.And this electrode pair vitamin C has fast response time (2 s), wide linear range (6.0 * 10
-8-1 * 10
-3M, significant sensitivity (187.3 mA M
1Cm
2), (0.024 μ M, low operating potential (0 V), (50 times analytical standard departs from 0.48% to excellent operation stability for low detectability; 13 standard deviations 0.56%) and storage stability (non-activity loss in 15 days; Internal loss in 30 days still had 80.05% biologically active in 2.6%, 60 day).The noiseless activity of most of materials.In addition, commercial beverage and vegetables are carried out the VC detection, good result shows that the application of this bio-sensing combination electrode has feasibility and operability.
Claims (1)
1. the preparation method of an ascorbic acid oxidase electrochemica biological sensing composite modified electrode, it is characterized in that: it may further comprise the steps:
(1), earlier with 1.42 mg 3; 4-enedioxy thiophene, 2.93 mg sarcosyls, 1 mg percent by weight, 0.1 wt% carbon nanometer tube dispersion liquid, 2.36 mg 1-ethyls-3-methylimidazole ethyl-sulfate ultrasonic dissolution mix at last and obtain electrolyte solution in the deionized water of 1 milliliter of cumulative volume;
(2), in above-mentioned electrolyte solution, fed nitrogen 20 minutes, adopt conventional three-electrode system, 1.1 V constant potential polymerizations 60 seconds, a step makes gathered (3,4-enedioxy thiophene) nano thin-film composite modified electrode, whole process is carried out under nitrogen protection;
(3), after composite modified electrode preparation finishes, electrode is taken out from electrolyte solution, use deionized water rinsing, then drying for standby;
(4), with 0.3g L
-1Ascorbic acid oxidase damping fluid 5 μ L drip to be coated in and gather (3,4-enedioxy thiophene) nano thin-film composite modified electrode surface, and air dry is subsequent use;
(5), with 4% carbon nanometer tube dispersion liquid, 5% Nafion solution, 1-butyl-3-methylimidazole hexafluorophosphoric acid hydrophobic nonionic liquid by volume 1:1 the three is mixed, ultrasonic preparation obtains 0.6 milliliter of carbon nanometer tube of cumulative volume-Nafion-hydrophobic nonionic liquid complex liquid;
(6), with carbon nanometer tube-Nafion-hydrophobic nonionic liquid complex liquid cover contain enzyme gather (3; 4-enedioxy thiophene) nano thin-film composite modified electrode surface, air dry obtains high-performance ascorbic acid oxidase electrochemica biological sensing composite modified electrode.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103018294A (en) * | 2012-10-29 | 2013-04-03 | 江西科技师范大学 | Method for preapraing electrochemical sensor for rapidly and selectively measuring maleic hydrazide in crops |
| CN103233256A (en) * | 2013-04-07 | 2013-08-07 | 浙江清华长三角研究院 | Preparation method of PEDOT/bmim[PF6] conductive ionic liquid polymer gas-sensitive film sensor |
| CN115201302A (en) * | 2022-07-04 | 2022-10-18 | 青岛科技大学 | Electrochemical sensor and preparation method and application thereof |
-
2011
- 2011-10-20 CN CN2011103201600A patent/CN102507691A/en active Pending
Non-Patent Citations (6)
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103018294A (en) * | 2012-10-29 | 2013-04-03 | 江西科技师范大学 | Method for preapraing electrochemical sensor for rapidly and selectively measuring maleic hydrazide in crops |
| CN103018294B (en) * | 2012-10-29 | 2014-08-20 | 江西科技师范大学 | Method for preapraing electrochemical sensor for rapidly and selectively measuring maleic hydrazide in crops |
| CN103233256A (en) * | 2013-04-07 | 2013-08-07 | 浙江清华长三角研究院 | Preparation method of PEDOT/bmim[PF6] conductive ionic liquid polymer gas-sensitive film sensor |
| CN103233256B (en) * | 2013-04-07 | 2016-05-04 | 浙江清华长三角研究院 | A kind of PEDOT/bmim[PF6] preparation method of conductive ion liquid polymers air-sensitive film sensor |
| CN115201302A (en) * | 2022-07-04 | 2022-10-18 | 青岛科技大学 | Electrochemical sensor and preparation method and application thereof |
| CN115201302B (en) * | 2022-07-04 | 2024-02-02 | 青岛科技大学 | Electrochemical sensor and preparation method and application thereof |
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Application publication date: 20120620 |