CN107490609A - Acetamiprid aptamers electrochemical sensor based on mesoporous silicon dioxde film - Google Patents
Acetamiprid aptamers electrochemical sensor based on mesoporous silicon dioxde film Download PDFInfo
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/301—Reference electrodes
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/38—Cleaning of electrodes
Abstract
Integration of the present invention by aptamers door control system with the vertical orderly mesoporous silicon dioxde film (MSF) for being grown in indium-tin oxide electrode (ITO) surface, builds a simple, sensitive electrochemical aptamer sensor platform.On this aptamer sensor platform, aptamers can be effectively adsorbed in amidized MSF surfaces by non-covalent electrostatic attraction, be used as preferable grid material, to control the closing and release of the electrochemistry reagent methylene blue captured in MSF holes.In the presence of Acetamiprid, the specific binding of aptamers and Acetamiprid can trigger the unlatching in hole, discharge methylene blue, reduce the current signal of detection.This electrochemical aptamer sensor design is simple, easy to operate, can realize quick, the sensitive detection to Acetamiprid.
Description
Technical field
The present invention relates to residues of pesticides field, is adapted in particular to a kind of electrochemistry based on MSFs modified electrodes
The preparation of body sensor and the detection of Acetamiprid.
Background technology
Agricultural chemicals typically has immunotoxicity, neurotoxicity, genetic toxicity and three causing toxicity etc., in order to improve and ensure
Food quality and life security, the highly sensitive detection to residues of pesticides are increasingly valued by people.Residues of pesticides are examined at present
The method of survey mainly has gas-chromatography, high performance liquid chromatography, Chromatography/Mass Spectrometry GC-MS, Capillary Electrophoresis, surface-enhanced Raman
Spectroscopic methodology, immunoassay, biology sensor etc..Gas-chromatography, high performance liquid chromatography and Chromatography/Mass Spectrometry GC-MS, separation effect
Can high, high sensitivity, but expensive equipment and being not easy realizes miniaturization, and the pretreatment process of sample is relatively complicated；Capillary
Electrophoresis then has the advantages that fast more clastotype, efficiency high, analyze speed, reagent and sample consumption are few, but capillary diameter is small,
Light path is short, poor reproducibility；SERS method has very high sensitivity, but reappearance and stability are poor.Electrochemistry
Sensor is with its low-power consumption, high sensitivity, high accuracy, strong antijamming capability, the range of linearity are wide and excellent repeatability, stability
Etc. characteristic, the every field such as biology, environment, food have been widely applied to.
The content of the invention
The purpose of the present invention is to provide one kind the shortcomings that being directed in above-mentioned Detecting Pesticide with aptamers door control system
With the integration of the vertical orderly mesoporous silicon dioxde film (MSF) that is grown in indium-tin oxide electrode (ITO) surface, an electricity is built
Chemical aptamer sensor platform, for quick, the Sensitive Detection to Acetamiprid.
The technical scheme is that：A kind of electrochemical method for detecting Acetamiprid agricultural chemicals, is concretely comprised the following steps：Graphite oxide
The preparation of alkene：6.8 g potassium permanganate are weighed, are added slowly with stirring the 120 mL concentrated sulfuric acids and 14 mL phosphoric acid；Add 1.0 g
Graphite powder, keep reaction temperature to be less than 20 DEG C, stir 15 min, afterwards add to mixed liquor in three-necked flask dropwise, at 50 DEG C
Continuous uniform stirs 12 h in water-bath；Add 140 mL frozen water to be allowed to cool down, be slowly added to 3.0 mL 30% hydrogen peroxide, now
Solution is in glassy yellow, and 2 h are ultrasonically treated after being cooled to room temperature, is then centrifuged with 6000 r/s rotating speed, supernatant is removed, with steaming
Distilled water washing is precipitated to neutrality, is placed at 60 DEG C and dries 12 h, obtains graphene oxide；
Gold-poly- 3,4- ethene dioxythiophenes/redox graphene（Au-PEDOT/rGO）The preparation of nano composite material：In room
While stirring by 5 mL 3,4- ethene dioxythiophenes under temperature（EDOT）（22.5 mmol/L）Ethanol solution pour into 70 mL
HAuCl4The aqueous solution（0.65 mmol/L）In, mixture is immediately turned into navy blue.In this process, EDOT（3,4- ethylene dioxies
Thiophene）Monomer is oxidized and polymerize to obtain PEDOT, and HAuCl4Ion is reduced as oxidant, while forms AuNP.Will
Reaction is kept stirring for 4 hours, then adds 5 mL GO aqueous dispersions（0.5 mg/mL）, obtain mixture solution（It is expressed as Au-
PEDOT/GO）；The Au-PEDOT/GO of acquisition is ultrasonically treated 2 hours, makes Au-PEDOT and GO fully dispersed；Then, quick
By 5.5 mL NaBH under stirring4The aqueous solution（0.16 mol/L）It is slowly dropped in mixture, and reaction system is kept stirring for
6 hours；Finally, by the mixture of acquisition（Au-PEDOT/rGO）Centrifuge and washed repeatedly with water and ethanol, then by solid point
It is dispersed in 50 mL deionized waters for future use；
The preparation of electrochemical aptamer sensor：ITO electrode is cleaned by ultrasonic 15 in acetone, second alcohol and water respectively first
Min, by 10 μ L Au-PEDOT/rGO drop coatings in the ITO electrode surface of cleaning, and dried in 45 DEG C of baking oven；Will modification
The ITO electrode of base material is immersed containing 35 mL water, 15 mL ethanol, 0.08 g CTAB, 5 μ L ammonia spirits（25%）With
In 40 μ L TEOS mixture solution, allow MSFs to be grown 48 hours under 60 DEG C, quiescent conditions, by ITO electrode ethanol and
Water rinses, and is dried overnight at 60 DEG C；The MSFs electrodes being modified are immersed into the ethanol containing 0.1 mol/L HCl under agitation
In 5 minutes, to remove ctab surface activating agent.Finally, ITO electrode MSFs being modified immerses the second that 4 mL contain 5%APTES
12 hours in alcoholic solution, gently shake, obtain NH2- MSFs, methylene blue concentration is dipped in as 10-5Mol/L Tris-
HCl solution（10 mmol/L, pH=7.4）In, and shaken overnight at room temperature；Then, by 10 μ L 10-8Mol/L adaptation
Liquid solution is added drop-wise to the MSFs modified electrodes surface of load methylene blue, is incubated 2 hours at room temperature, obtains aptamers end-blocking
MSFs modified electrodes；After being incubated with aptamers, by modified ITO electrode with 10 mmol/L Tris-HCl buffer solutions（pH=7.4）
Wash to remove non-specific adsorption；At room temperature, the 10 μ L of MSFs modified electrodes immersion of aptamers end-blocking are contained a certain amount of
2 hours in the cushioning liquid of Acetamiprid；Finally, thoroughly washed by using buffer solution suitable with Acetamiprid specific binding to remove
Part.
A kind of electrochemical method for detecting Acetamiprid, it is specific rapid as follows：
（1）It is working electrode by the ITO of above-mentioned modification in the electrolytic cell of the PBS solution containing 10 mL pH 7.0, Ag/
AgCl electrodes are reference electrode, and platinum electrode is to electrode；Experiment is carried out on CHI842C Electrochemical Comprehensive Testers, and its is attached
Computer software be made for the collection and processing of experimental data；Differential pulse scanning is carried out in -0.5~0.1 V potential ranges,
Differential pulse voltammetry voltammogram is recorded, measurement is not added with peak point current during AcetamipridIp 0 ；According to said method, the pyridine worm of various concentrations is added
Amidine, you can obtain corresponding peak point currentIp x , calculate peak current difference △Ip（△Ip=I p0 - I px ）；△Ip/Ip 0 With pyridine worm
The logarithm of amidine concentrationlgcGood linear relationship, linear equation △ is presentedIp/Ip 0 =0.49258+0.05731 lgc, c are
Concentration, unit are mol/L, △IpIt is peak current difference, unit is mA, linearly dependent coefficient R=0.99477, Acetamiprid concentration
The range of linearity is 10-8 mol/L~10-13Mol/L, detection of the sensor to Acetamiprid agricultural chemicals are limited to 3.4 × 10-14mol/L；
（2）With reference to above-mentioned linear relationship, the Acetamiprid sample of unknown concentration is measured, calculates Acetamiprid concentration：Containing
In the electrolytic cell for having 10 mL pH 7.0 PBS solution, using the ITO of above-mentioned modification as working electrode, Ag/AgCl electrodes are ginseng
Than electrode, platinum electrode is to electrode；Experiment is carried out on CHI842C Electrochemical Comprehensive Testers, its attached computer software
It is made for the collection and processing of experimental data；A certain amount of solution to be measured is added, difference is carried out in -0.5~0.1 V potential ranges
Pulse scans, and records differential pulse voltammetry voltammogram, obtains peak point currentI p, willI pAbove-mentioned equation is substituted into, can ask and calculate pyridine in prepare liquid
The concentration of worm amidine.
The beneficial effects of the present invention are：The present invention is by aptamers door control system with being grown in the vertical of ITO electrode surface
The ingenious integration of orderly MSFs is prepared for a simple, light, unmarked electrochemical aptamer sensor platform；Use adaptation
Body is as grid material, to control the obstruction of MSFs mesoporous Methylene Blue and release；Based between aptamers and target substance
High-affinity and specificity, aptamers only adsorbed by non-covalent electrostatic attraction easy to operate in MSFs surfaces, this method
And without any complicated chemical modification；Prepared electrochemical aptamer sensor shows high sensitivity and good to Acetamiprid
Good selectivity, test result indicates that, the electrochemical aptamer sensor proposed, have in trace Pesticides Testing and potentially should
Use prospect.
Brief description of the drawings：
DPV signal intensity stacking charts under Fig. 1 difference Acetamiprid concentration
Wherein, 1-0 mol/L, 2-10-13Mol/L, 3-10-12Mol/L, 4-10-11Mol/L,
5—10-10Mol/L, 6-10-9Mol/L, 7-10-9mol/L；
Ratio and lg of Fig. 2 peak current differences of the present invention with current value when being not added with AcetamipridcLinear relationship chart.
Embodiment：
For a better understanding of the present invention, technical scheme is described in detail with instantiation below, but it is of the invention
It is not limited thereto.
Embodiment 1
1. the preparation of graphene oxide
6.8 g potassium permanganate are weighed, are added slowly with stirring the 120 mL concentrated sulfuric acids and 14 mL phosphoric acid；Add 1.0 g graphite
Powder, keep reaction temperature to be less than 20 DEG C, stir 15 min, afterwards add to mixed liquor in three-necked flask dropwise, in 50 DEG C of water-baths
Middle continuous uniform stirs 12 h；Add 140 mL frozen water to be allowed to cool down, be slowly added to 3.0 mL 30% hydrogen peroxide, now solution
In glassy yellow, 2 h are ultrasonically treated after being cooled to room temperature, is then centrifuged with 6000 r/s rotating speed, is removed supernatant, use distilled water
Washing is precipitated to neutrality, is placed at 60 DEG C and dries 12 h, obtains graphene oxide；
2. the preparation of Au-PEDOT/rGO nano composite materials
Gold-poly- 3,4- ethene dioxythiophenes/redox graphene（Au-PEDOT/rGO）The preparation of nano composite material：In room
While stirring by 5 mL 3,4- ethene dioxythiophenes under temperature（EDOT）（22.5 mmol/L）Ethanol solution pour into 70 mL
HAuCl4The aqueous solution（0.65 mmol/L）In, mixture is immediately turned into navy blue.In this process, EDOT（3,4- ethylene dioxies
Thiophene）Monomer is oxidized and polymerize to obtain PEDOT, and HAuCl4Ion is reduced as oxidant, while forms AuNP.Will
Reaction is kept stirring for 4 hours, then adds 5 mL GO aqueous dispersions（0.5 mg/mL）, obtain mixture solution（It is expressed as Au-
PEDOT/GO）；The Au-PEDOT/GO of acquisition is ultrasonically treated 2 hours, makes Au-PEDOT and GO fully dispersed；Then, quick
By 5.5 mL NaBH under stirring4The aqueous solution（0.16 mol/L）It is slowly dropped in mixture, and reaction system is kept stirring for
6 hours；Finally, by the mixture of acquisition（Au-PEDOT/rGO）Centrifuge and washed repeatedly with water and ethanol, then by solid point
It is dispersed in 50 mL deionized waters for future use；
3. the preparation of electrochemical aptamer sensor
ITO electrode is cleaned by ultrasonic 15 min in acetone, second alcohol and water respectively first, by 10 μ L Au-PEDOT/rGO drop coatings
In the ITO electrode surface of cleaning, and dried in 45 DEG C of baking oven；The ITO electrode for having modified base material is immersed and contained
35 mL water, 15 mL ethanol, 0.08 g CTAB, 5 μ L ammonia spirits（25%）In 40 μ L TEOS mixture solution, allow
MSFs is grown 48 hours under 60 DEG C, quiescent conditions, and ITO electrode is rinsed with second alcohol and water, and is dried overnight at 60 DEG C；Will
The electrode that MSFs is modified immerses in the ethanol containing 0.1 mol/L HCl 5 minutes under agitation, to remove ctab surface activity
Agent.Finally, ITO electrode MSFs being modified is immersed in the ethanol solution that 4 mL contain 5%APTES 12 hours, is gently shaken, is obtained
To NH2- MSFs, methylene blue concentration is dipped in as 10-5Mol/L Tris-HCl solution（10 mmol/L, pH=7.4）
In, and shaken overnight at room temperature；Then, by 10 μ L 10-8Mol/L adaptation liquid solution is added drop-wise to load methylene blue
MSFs modified electrodes surface, it is incubated 2 hours at room temperature, obtains the MSFs modified electrodes of aptamers end-blocking；It is incubated with aptamers
Afterwards, by modified ITO electrode with 10 mmol/L Tris-HCl buffer solutions（pH=7.4）Wash to remove non-specific adsorption；
At room temperature, the 10 μ L of MSFs modified electrodes immersion of aptamers end-blocking are contained in the cushioning liquid of a certain amount of Acetamiprid 2 hours；
Finally, thoroughly washed by using buffer solution to remove the aptamers with Acetamiprid specific binding.
4. utilize the content of above-mentioned aptamer sensor detection Acetamiprid
（1）It is working electrode by the ITO of above-mentioned modification in the electrolytic cell of the PBS solution containing 10 mL pH 7.0, Ag/
AgCl electrodes are reference electrode, and platinum electrode is to electrode；Experiment is carried out on CHI842C Electrochemical Comprehensive Testers, and its is attached
Computer software be made for the collection and processing of experimental data；Differential pulse scanning is carried out in -0.5~0.1 V potential ranges,
Differential pulse voltammetry voltammogram is recorded, measurement is not added with peak point current during AcetamipridIp 0 ；According to said method, the pyridine worm of various concentrations is added
Amidine, you can obtain corresponding peak point currentIp x , calculate peak current difference △Ip（△Ip=I p0 - I px ）；△IpIt is dense with Acetamiprid
The logarithm of degreelgcGood linear relationship, linear equation △ is presentedIp/Ip 0 =0.49258+0.05731 lgc, c are concentration,
Unit is mol/L, △IpPeak current difference, unit is mA, linearly dependent coefficient R=0.99477, Acetamiprid concentration it is linear
Scope is 10-8 mol/L~10-13Mol/L, detection of the sensor to Acetamiprid agricultural chemicals are limited to 3.4 × 10-14mol/L；
（2）With reference to above-mentioned linear relationship, the Acetamiprid sample of unknown concentration is measured, calculates Acetamiprid concentration：Containing
In the electrolytic cell for having 10 mL pH 7.0 PBS solution, using the ITO of above-mentioned modification as working electrode, Ag/AgCl electrodes are ginseng
Than electrode, platinum electrode is to electrode；Experiment is carried out on CHI842C Electrochemical Comprehensive Testers, its attached computer software
It is made for the collection and processing of experimental data；A certain amount of solution to be measured is added, difference is carried out in -0.5~0.1 V potential ranges
Pulse scans, and records differential pulse voltammetry voltammogram, obtains peak point currentI p, willI pAbove-mentioned equation is substituted into, can ask and calculate pyridine in prepare liquid
The concentration of worm amidine.
Claims (2)
1. a kind of preparation for the electrochemical aptamer sensor for detecting Acetamiprid, it is characterised in that concretely comprise the following steps：First will
ITO electrode is cleaned by ultrasonic 15 min in acetone, second alcohol and water respectively, by 10 μ L Au-PEDOT/rGO drop coatings cleaning ITO
On electrode surface, and dried in 45 DEG C of baking oven；The ITO electrode for having modified base material is immersed containing 35 mL water, 15
ML ethanol, 0.08 g CTAB, 5 μ L ammonia spirits（25%）In 40 μ L TEOS mixture solution, allow MSFs 60 DEG C,
After being grown 48 hours under quiescent conditions, ITO electrode is rinsed with second alcohol and water, and is dried overnight at 60 DEG C；MSFs is modified
Electrode immerse under agitation in the ethanol containing 0.1 mol/L HCl 5 minutes, to remove ctab surface activating agent；Finally, will
The ITO electrode that MSFs is modified is immersed in the ethanol solution that 4 mL contain 5%APTES 12 hours, is gently shaken, to ensure MSFs's
Surface can further use aminofunctional；After the ITO electrode amination that MSFs is modified, it is 10 to immerse methylene blue concentration-5
Mol/L Tris-HCl solution（10 mmol/L, pH=7.4）In, and shaken overnight at room temperature；Then, by 10 μ L 10-8
Mol/L adaptation liquid solution is added drop-wise to the MSFs modified electrodes surface of load methylene blue, is incubated 2 hours, obtains at room temperature
The MSFs modified electrodes of aptamers end-blocking；After being incubated with aptamers, modified ITO electrode is delayed with 10 mmol/L Tris-HCl
Fliud flushing（pH=7.4）Wash to remove non-specific adsorption；At room temperature, the MSFs modified electrodes of aptamers end-blocking are immersed 10
μ L contain in the cushioning liquid of a certain amount of Acetamiprid 2 hours；Finally, washed with buffer solution and remove what is specifically bound with Acetamiprid
Aptamers.
2. Acetamiprid is detected according to the electrochemical aptamer sensor prepared by claim 1, it is characterised in that specific steps
For：
（1）It is working electrode by the ITO of above-mentioned modification in the electrolytic cell of the PBS solution containing 10 mL pH 7.0, Ag/
AgCl electrodes are reference electrode, and platinum electrode is to electrode；Experiment is carried out on CHI842C Electrochemical Comprehensive Testers, and its is attached
Computer software be made for the collection and processing of experimental data；Differential pulse scanning is carried out in -0.5~0.1 V potential ranges,
Differential pulse voltammetry voltammogram is recorded, measurement is not added with peak point current during AcetamipridIp 0 ；According to said method, the pyridine worm of various concentrations is added
Amidine, you can obtain corresponding peak point currentIp x , calculate peak current difference △Ip（△Ip=I p0 -I px ）；DIp/ Ip 0 With Acetamiprid
The logarithm of concentrationlgcGood linear relationship, linear equation △ is presentedIp/Ip 0 =0.49258+0.05731lgc, c are dense
Degree, unit is mol/L, △IpIt is peak current difference, unit is mA, linearly dependent coefficient R=0.99477, the line of Acetamiprid concentration
Property scope be 10-8 mol/L~10-13Mol/L, detection of the sensor to Acetamiprid agricultural chemicals are limited to 3.4 × 10-14mol/L；
（2）With reference to above-mentioned linear relationship, the Acetamiprid sample of unknown concentration is measured, calculates Acetamiprid concentration：Containing
In the electrolytic cell for having 10 mL pH 7.0 PBS solution, using the ITO of above-mentioned modification as working electrode, Ag/AgCl electrodes are ginseng
Than electrode, platinum electrode is to electrode；Experiment is carried out on CHI842C Electrochemical Comprehensive Testers, its attached computer software
It is made for the collection and processing of experimental data；A certain amount of solution to be measured is added, difference is carried out in -0.5~0.1 V potential ranges
Pulse scans, and records differential pulse voltammetry voltammogram, obtains peak point currentI p, willI pAbove-mentioned equation is substituted into, can ask and calculate pyridine in prepare liquid
The concentration of worm amidine.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110455897A (en) * | 2019-08-29 | 2019-11-15 | 济南大学 | One kind being based on SiO2Carrier Sensitive Detection Hg2+Release type electrochemical aptamer sensor building |
| CN113447553A (en) * | 2021-06-21 | 2021-09-28 | 同济大学 | Non-immobilized electrochemical sensor based on signal probe packaging release and application thereof |
| CN114624298A (en) * | 2020-12-12 | 2022-06-14 | 中国科学院大连化学物理研究所 | Method for detecting ochratoxin A by electrochemical paper chip based on aptamer gating effect |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110455897A (en) * | 2019-08-29 | 2019-11-15 | 济南大学 | One kind being based on SiO2Carrier Sensitive Detection Hg2+Release type electrochemical aptamer sensor building |
| CN110455897B (en) * | 2019-08-29 | 2021-07-02 | 济南大学 | Based on SiO2Sensitive detection of Hg by carrier2+Construction of the Release electrochemical aptamer sensor |
| CN114624298A (en) * | 2020-12-12 | 2022-06-14 | 中国科学院大连化学物理研究所 | Method for detecting ochratoxin A by electrochemical paper chip based on aptamer gating effect |
| CN114624298B (en) * | 2020-12-12 | 2023-05-05 | 中国科学院大连化学物理研究所 | Method for detecting ochratoxin A by using electrochemical paper chip based on aptamer gating effect |
| CN113447553A (en) * | 2021-06-21 | 2021-09-28 | 同济大学 | Non-immobilized electrochemical sensor based on signal probe packaging release and application thereof |
| CN113447553B (en) * | 2021-06-21 | 2022-09-20 | 同济大学 | Non-immobilized electrochemical sensor based on signal probe packaging release and application thereof |
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| Publication number | Publication date |
|---|---|
| CN107490609B (en) | 2019-11-01 |
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