CN102393411A - Preparation method of electrochemical sensor for detecting molecular imprinting of organophosphorus pesticide residue - Google Patents
Preparation method of electrochemical sensor for detecting molecular imprinting of organophosphorus pesticide residue Download PDFInfo
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- CN102393411A CN102393411A CN2011103184592A CN201110318459A CN102393411A CN 102393411 A CN102393411 A CN 102393411A CN 2011103184592 A CN2011103184592 A CN 2011103184592A CN 201110318459 A CN201110318459 A CN 201110318459A CN 102393411 A CN102393411 A CN 102393411A
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Abstract
The invention discloses a preparation method of an electrochemical sensor for detecting molecular imprinting of organophosphorus pesticide residue. The method comprises the following steps of: firstly preparing a solution from the raw molecules of organophosphorus pesticide serving as imprinting molecules and a functional monomer and an electrolyte; depositing the molecular imprinting polymer on the surface of a working electrode by an electrochemical polymerization method so as to finish the preparation of a molecular imprinting sensitive film; and removing the imprinting molecules of the organophosphorus pesticide from the molecular imprinting sensitive film by a proper elution method and a proper eluent so as to finish the preparation of the electrochemical sensor. The method disclosed by the invention has the advantages of fast response, high sensitivity and good selectivity; and meanwhile, the detection method is simple and quick, has strong adaptability after being popularized and particularly strong universality in pesticide detection, and has a wide market prospect.
Description
Technical field
The invention belongs to the Pesticides Testing technical field, relate to a kind of preparation method of electrochemical sensor, be specifically related to a kind of preparation method who detects the electrochemical sensor of organophosphorus pesticide molecular engram.
Background technology
Organophosphorus pesticide is a kind of neurovirulent high-efficient broad-spectrum insecticide that has, and commonly used have kinds more than 60 such as Rogor, metrifonate, DDVP, demeton, parathion, malathion.Organophosphorus pesticide unstable being prone in physical environment decomposed, and is prone to decomposed by enzyme behind the entering biosome, and the residence time is shorter in food; Be difficult for causing slow poisoning; But can often cause acute poisoning, can cause muscular tremor, spasm, elevation of blood pressure, palpitating speed etc. when serious, even cause stupor dead; Therefore be directly connected to the security of food for the detection control of the organophosphorus pesticide of agricultural product, healthy closely bound up with people.The material that the molecular engram electrochemical sensor is mainly used in micro-trace detects, and it is quick to have analyzing and testing, simple to operate; Result reliability is good, and high temperature resistant, high pressure, acid, alkali and organic solvent are widely used in the food Detecting Pesticide; Environment measuring, fields such as analytical test.
Molecular imprinting (Molecular imprinting technique is called for short MIT) is multidisciplinary cross products such as high polymer chemistry, biological chemistry and materials chemistry; The molecular engram electrochemical sensing combines the two advantage of molecular imprinting and electrochemical analysis method; Utilize molecularly imprinted polymer can realize that as the electrochemical sensor of sensitive material preparation the express-analysis of target substance trace trace detects, and occupies an important position in the analyzing and testing field.This invention is carried out preliminary exploration to the preparation of the molecular engram sensitive membrane of organophosphorus insecticide, has successfully prepared the molecular engram electrochemical sensor of organophosphorus insecticide, can satisfy the needs of practical application, has certain application development prospect.
Summary of the invention
The purpose of this invention is to provide a kind of preparation method who detects the electrochemical sensor of organophosphorus pesticide molecular engram; Go out to prepare the top condition of electrochemical sensor sensitive membrane through experimental exploring; The ratio of research work electrode, electrochemical polymerization solvent, electrochemical polymerization, the former medicine microsphere of organophosphorus pesticide and function monomer, electrochemical polymerization time etc. influence the factor of electrochemical sensor preparation; Make the electrochemical sensor of preparation have the advantage that response is quick, highly sensitive and selectivity is good, can realize that quick on-the site analysis detects.
The technical scheme that the present invention adopted is, detects the preparation method of the electrochemical sensor of organophosphorus pesticide molecular engram, specifically implements according to following steps:
Step 1: with 0.05 micron-0.30 micron Alpha-alumina slurry polishing electrode; Adopt ethanol, acetone, the water cleaning electrode that heavily steams respectively, the electrode after cleaning is inserted KCl and K
3[Fe (CN)
6] mixed solution in, the stability that adopts the electrochemical cyclic voltammetry detecting electrode between 0-0.5V, the high purity water cleaning electrode;
Step 2: the function monomer o-aminophenol is dissolved in HClO
4In the solution, use the NaOH solution regulator solution pH value of 1mol/L to be 4-6;
Step 3: microsphere DDVP is joined in the function monomer solution that step 2 obtains, and the mol ratio of control function monomer and microsphere is 4: 1-6: 1;
Step 4: shading was left standstill after the logical nitrogen of the mixed solution that step 3 is obtained was removed oxygen;
Step 5: the electrode that inserting step 1 obtains in the mixed solution that step 4 obtains, adopt the electrochemical cyclic voltammetry sedimentation to prepare organophosphorus pesticide molecular engram electrochemical sensor sensitive membrane;
Step 6: adopt the microsphere in the sensitive membrane that solvent elution method elution step 5 obtains, obtain detecting the electrochemical sensor of organophosphorus pesticide molecular engram.
Characteristics of the present invention also are,
Working electrode in the step 1 is wherein selected glass-carbon electrode for use.
KCl in the step 1 wherein and K
3[Fe (CN)
6] volumetric molar concentration be respectively 0.1mol/L and 5mol/L.
The volumetric molar concentration of the function monomer o-aminophenol in the step 2 wherein is 0.005-0.01mol/L.
The volumetric molar concentration of the microsphere DDVP in the step 3 wherein is 0.02-0.08mol/L.
The logical nitrogen time of step 4 wherein is 10min, and 3h is left standstill in airtight shading.
Electrochemical cyclic voltammetry sedimentation in the step 5 wherein, electrochemical deposition current potential are 0-0.6V, and the scanning number of turns is 15 circles.
Eluant, eluent in the step 6 wherein, adopting volume ratio is 3: 1-5: the mixed solution of 1 acetonitrile and the dilution heat of sulfuric acid of 0.5mol/L, elution time are 12h.
The invention has the beneficial effects as follows,
1. the preparation of organophosphorus pesticide molecular engram electrochemical sensor: the molecular structure characteristics according to organophosphorus pesticide are selected suitable working electrode and function monomer and suitable electrochemical polymerization solvent, and preparation has highly sensitive molecularly imprinted polymer sensitive membrane.The ratio of the selection of the selection of research work electrode of the present invention, electrochemical polymerization choice of Solvent, function monomer, the selection of electrochemical polymerization, the former medicine microsphere of organophosphorus pesticide and function monomer, electrochemical polymerization time etc. influence the factor of molecular engram sensitive membrane performance; Explore best preparation condition; Prepare the sensitive membrane of molecular engram electrochemical sensor through suitable elution process, and then pass through electrochemical analysis method and realize that express-analysis detects the electrochemical sensor of organophosphorus pesticide residual quantity with molecular recognition performance.
2. organophosphorus pesticide molecular engram electrochemical sensing Study on Selectivity: the research electrochemical sensor is to the least concentration of the size of the response signal of different agricultural chemicals, target microsphere that sensor can detect and the mechanism of analyzing and testing thereof.
Technically, key problem in technology of the present invention is: to guarantee that at first organic phosphorous molecule imprinted electrochemical sensor has quick response to the target microsphere, and highly sensitive, can realize analyzing and testing to the residues of pesticides trace; Secondly the organic phosphorous molecule imprinted electrochemical sensor of preparation has good selectivity to the target microsphere, and response signal is eager to excel than other material, can reach the purpose of molecular recognition, realizes the detection of organophosphorus pesticide residual quantity trace.
Embodiment
Below in conjunction with embodiment the present invention is elaborated.
The present invention detects the preparation method of the electrochemical sensor of organophosphorus pesticide molecular engram, specifically implements according to following steps:
Step 1: the selection of electrode and pre-service: select for use glass-carbon electrode as working electrode, with 0.05 micron-0.30 micron Alpha-alumina slurry polishing; Adopt the ethanol that heavily steams, acetone, water is cleaning electrode respectively, and the electrode after cleaning is inserted KCl and K
3[Fe (CN)
6] mixed solution in, KCl and K
3[Fe (CN)
6] volumetric molar concentration be respectively 0.1mol/L and 5mol/L; The stability that adopts the electrochemical cyclic voltammetry detecting electrode between 0-0.5V, the high purity water cleaning electrode;
Step 2: the preparation of function monomer solution: according to volumetric molar concentration is 0.005-0.01mol/L, and a certain amount of function monomer o-aminophenol is dissolved in HClO
4In the solution, use the NaOH solution regulator solution pH value of 1mol/L to be 4-6;
Step 3: the selection of microsphere: according to volumetric molar concentration is 0.02-0.08mol/L, and a certain amount of microsphere DDVP is joined in the function monomer solution that step 2 obtains, and the mol ratio of control function monomer and microsphere is 4: 1-6: 1;
Step 4: shading was left standstill after the logical nitrogen of the mixed solution that step 3 is obtained was removed oxygen, and the logical nitrogen time is 10min, and 3h is left standstill in airtight shading;
Step 5: the electrode after inserting step 1 is handled, adopt the electrochemical cyclic voltammetry sedimentation to prepare organophosphorus pesticide molecular engram electrochemical sensor sensitive membrane; The electrochemical deposition current potential is 0-0.6V, and the scanning number of turns (sedimentation time) is 15 circles;
Step 6: the microsphere in the sensitive membrane that employing solvent elution method elution step 5 obtains; Adopting volume ratio is 3: 1-5: the mixed solution of 1 acetonitrile, the dilution heat of sulfuric acid of 0.5mol/L is an eluant, eluent wash-out microsphere; Elution time is 12h, obtains detecting organophosphorus pesticide molecular engram electrochemical sensor.
The organophosphorus pesticide molecular engram electrochemical sensor for preparing is in organophosphorus pesticide concentration 3.0 * 10
-7-1.5 * 10
-6Mol/L and 1.0 * 10
-6-1.0 * 10
-5In the mol/L scope, the sensor peak current changes with organophosphorus pesticide concentration linear.Linear equation is respectively: y=2.332x+4.3 * 10
-8, related coefficient is R2=0.9631; Y=0.3972x+6.2 * 10
-6, related coefficient is R2=0.9725, is limited to 4.2 * 10 under detecting
-8Mol/L.
Below the present invention will be described from the principle aspect:
(1) selection of working electrode and function monomer: the chemical constitution according to the organophosphorus pesticide microsphere is selected suitable working electrode and function monomer; Microsphere generally interacts through action and function monomers such as hydrogen bond, intermolecular force, electrostatic force; Therefore the amount of force of microsphere and function monomer is directly connected to the quality of molecular engram electrochemical sensor sensitive membrane preparation; The too little meeting of acting force causes the selection identity in the trace hole that molecular engram electrochemical sensor sensitive membrane forms behind wash-out poor; Acting force too greatly then causes microsphere to be difficult to that wash-out comes out from sensor sensing film; The molecular engram hole that causes forming very little, the electrochemical response signal is too for a short time not to reach the requirement that electrochemical analysis detects.
(2) preparation of function monomer and organophosphorus pesticide electrochemical polymerization solution: the galvanochemistry polymerization base fluid of the different mol ratio of preparation function monomer and organophosphorus pesticide microsphere; Add a certain amount of electrolyte; Feed nitrogen earlier and remove the oxygen in the solution; The regular hour is left standstill in the solution shading of preparation, make function monomer and microsphere fully act on the formation stabilized complex.
(3) preparation of organophosphorus pesticide molecular engram electrochemical sensor sensitive membrane: inquire into different electrochemical polymerization sedimentations; The electrochemical deposition current potential; Factors such as polymerization sedimentation time find best preparation condition to molecular engram electrochemical sensor sensitive membrane Effect on Performance, and it is big that the sensitive membrane of preparation has response signal; Response is sensitive, and selectivity is good.
(4) the microsphere wash-out in the organophosphorus pesticide molecular engram electrochemical sensor sensitive membrane: can on sensitive membrane, stay consistent in the molecular engram electrochemical sensor sensitive membrane behind the microsphere wash-out with the microsphere size; The hole that functional group arranges in certain sequence; And the quantity in the hole that forms directly has influence on the size of electrochemical response signal; Therefore select suitable elution process and eluting solvent to greatest extent the microsphere in the wash-out sensitive membrane form a fairly large number of molecular engram hole, reach the requirement of the response signal of electrochemical analysis detection.
(5) research of organophosphorus pesticide molecular engram electrochemical sensor practical application performance: the molecular engram electrochemical sensor of main research institute preparation is used for the detection of environment organophosphorus pesticide concentration; The selectivity of research in analyte detection process; Select for use different types of agricultural chemicals to carry out mark-on and reclaim experiment, confirm its actual Application feasibility.
Embodiment 1
(1) selecting glass-carbon electrode is electrode, with 0.05 micron-0.30 micron Alpha-alumina slurry polishing; Adopt the ethanol that heavily steams, acetone, water is cleaning electrode respectively; Electrode after handling is inserted 0.1mol/L KCl and 5mol/L K
3[Fe (CN)
6] mixed solution in; The stability that adopts the electrochemical cyclic voltammetry detecting electrode between 0-0.5V, the high purity water cleaning electrode;
(2) selecting o-aminophenol for use is function monomer, and it is dissolved in HClO
4In the solution, the concentration of o-aminophenol is 0.006mol/L, and using the NaOH solution regulator solution pH value of 1mol/L is 5;
(3) selecting DDVP is microsphere, and it is dissolved in the monomer solution, and controlling its concentration is 0.05mol/L, and the mol ratio that makes function monomer and microsphere is 5: 1;
(4) the logical nitrogen time is 10min, and 3h is left standstill in airtight shading;
(5) adopt the electrochemical cyclic voltammetry sedimentation to prepare organophosphorus pesticide molecular engram electrochemical sensor sensitive membrane, the electrochemical deposition current potential is 0-0.6V, and the scanning number of turns (sedimentation time) is 15 circles;
(6) dilution heat of sulfuric acid of employing acetonitrile, 0.5mol/L is that the mixed solution of 4: 1 (volume ratio) is an eluant, eluent wash-out microsphere, and elution time is 12h.
Embodiment 2
(1) selecting glass-carbon electrode is electrode, with 0.05 micron-0.30 micron Alpha-alumina slurry polishing; Adopt the ethanol that heavily steams, acetone, water is cleaning electrode respectively; Electrode after handling is inserted 0.1mol/L KCl and 5mol/L K
3[Fe (CN)
6] mixed solution in; The stability that adopts the electrochemical cyclic voltammetry detecting electrode between 0-0.5V, the high purity water cleaning electrode;
(2) selecting o-aminophenol for use is function monomer, and it is dissolved in HClO
4In the solution, the concentration of o-aminophenol is 0.005mol/L, and using the NaOH solution regulator solution pH value of 1mol/L is 4;
(3) selecting DDVP is microsphere, and it is dissolved in the monomer solution, and controlling its concentration is 0.02mol/L, and the mol ratio that makes function monomer and microsphere is 4: 1;
(4) the logical nitrogen time is 10min, and 3h is left standstill in airtight shading;
(5) adopt the electrochemical cyclic voltammetry sedimentation to prepare organophosphorus pesticide molecular engram electrochemical sensor sensitive membrane, the electrochemical deposition current potential is 0-0.6V, and the scanning number of turns (sedimentation time) is 15 circles;
(6) dilution heat of sulfuric acid of employing acetonitrile, 0.5mol/L is that the mixed solution of 3: 1 (volume ratio) is an eluant, eluent wash-out microsphere, and elution time is 12h.
Embodiment 3
(1) selecting glass-carbon electrode is electrode, with 0.05 micron-0.30 micron Alpha-alumina slurry polishing; Adopt the ethanol that heavily steams, acetone, water is cleaning electrode respectively; Electrode after handling is inserted 0.1mol/L KCl and 5mol/L K
3[Fe (CN)
6] mixed solution in; The stability that adopts the electrochemical cyclic voltammetry detecting electrode between 0-0.5V, the high purity water cleaning electrode;
(2) selecting o-aminophenol for use is function monomer, and it is dissolved in HClO
4In the solution, the concentration of o-aminophenol is 0.01mol/L, and using the NaOH solution regulator solution pH value of 1mol/L is 6;
(3) selecting DDVP is microsphere, and it is dissolved in the monomer solution, and controlling its concentration is 0.08mol/L, and the mol ratio that makes function monomer and microsphere is 6: 1;
(4) the logical nitrogen time is 10min, and 3h is left standstill in airtight shading;
(5) adopt the electrochemical cyclic voltammetry sedimentation to prepare organophosphorus pesticide molecular engram electrochemical sensor sensitive membrane, the electrochemical deposition current potential is 0-0.6V, and the scanning number of turns (sedimentation time) is 15 circles;
(6) dilution heat of sulfuric acid of employing acetonitrile, 0.5mol/L is that the mixed solution of 5: 1 (volume ratio) is an eluant, eluent wash-out microsphere, and elution time is 12h.
Embodiment 4
(1) selecting glass-carbon electrode is electrode, with 0.05 micron-0.30 micron Alpha-alumina slurry polishing; Adopt the ethanol that heavily steams, acetone, water is cleaning electrode respectively; Electrode after handling is inserted 0.1mol/L KCl and 5mol/L K
3[Fe (CN)
6] mixed solution in; The stability that adopts the electrochemical cyclic voltammetry detecting electrode between 0-0.5V, the high purity water cleaning electrode;
(2) selecting o-aminophenol for use is function monomer, and it is dissolved in HClO
4In the solution, the concentration of o-aminophenol is 0.007mol/L, and using the NaOH solution regulator solution pH value of 1mol/L is 6;
(3) selecting DDVP is microsphere, and it is dissolved in the monomer solution, and controlling its concentration is 0.06mol/L, and the mol ratio that makes function monomer and microsphere is 4: 1;
(4) the logical nitrogen time is 10min, and 3h is left standstill in airtight shading;
(5) adopt the electrochemical cyclic voltammetry sedimentation to prepare organophosphorus pesticide molecular engram electrochemical sensor sensitive membrane, the electrochemical deposition current potential is 0-0.6V, and the scanning number of turns (sedimentation time) is 15 circles;
(6) dilution heat of sulfuric acid of employing acetonitrile, 0.5mol/L is that the mixed solution of 5: 1 (volume ratio) is an eluant, eluent wash-out microsphere, and elution time is 12h.
Claims (8)
1. detect the preparation method of the electrochemical sensor of organophosphorus pesticide molecular engram, it is characterized in that, specifically implement according to following steps:
Step 1: with 0.05 micron-0.30 micron Alpha-alumina slurry polishing electrode; Adopt ethanol, acetone, the water cleaning electrode that heavily steams respectively, the electrode after cleaning is inserted KCl and K
3[Fe (CN)
6] mixed solution in, the stability that adopts the electrochemical cyclic voltammetry detecting electrode between 0-0.5V, the high purity water cleaning electrode;
Step 2: the function monomer o-aminophenol is dissolved in HClO
4In the solution, use the NaOH solution regulator solution pH value of 1mol/L to be 4-6;
Step 3: microsphere DDVP is joined in the function monomer solution that step 2 obtains, and the mol ratio of control function monomer and microsphere is 4: 1-6: 1;
Step 4: shading was left standstill after the logical nitrogen of the mixed solution that step 3 is obtained was removed oxygen;
Step 5: the electrode that inserting step 1 obtains in the mixed solution that step 4 obtains, adopt the electrochemical cyclic voltammetry sedimentation to prepare organophosphorus pesticide molecular engram electrochemical sensor sensitive membrane;
Step 6: adopt the microsphere in the sensitive membrane that solvent elution method elution step 5 obtains, obtain detecting the electrochemical sensor of organophosphorus pesticide molecular engram.
2. the preparation method of the electrochemical sensor of detection organophosphorus pesticide molecular engram according to claim 1 is characterized in that the working electrode in the described step 1 is selected glass-carbon electrode for use.
3. the preparation method of the electrochemical sensor of detection organophosphorus pesticide molecular engram according to claim 1 is characterized in that, KCl in the described step 1 and K
3[Fe (CN)
6] volumetric molar concentration be respectively 0.1mol/L and 5mol/L.
4. the preparation method of the electrochemical sensor of detection organophosphorus pesticide molecular engram according to claim 1 is characterized in that, the volumetric molar concentration of the function monomer o-aminophenol in the described step 2 is 0.005-0.01mol/L.
5. the preparation method of the electrochemical sensor of detection organophosphorus pesticide molecular engram according to claim 1 is characterized in that, the volumetric molar concentration of the microsphere DDVP in the described step 3 is 0.02-0.08mol/L.
6. the preparation method of the electrochemical sensor of detection organophosphorus pesticide molecular engram according to claim 1 is characterized in that, the logical nitrogen time of described step 4 is 10min, and 3h is left standstill in airtight shading.
7. the preparation method of the electrochemical sensor of detection organophosphorus pesticide molecular engram according to claim 1; It is characterized in that; Electrochemical cyclic voltammetry sedimentation in the described step 5, electrochemical deposition current potential are 0-0.6V, and the scanning number of turns is 15 circles.
8. the preparation method of the electrochemical sensor of detection organophosphorus pesticide molecular engram according to claim 1; It is characterized in that; Eluant, eluent in the described step 6; Adopting volume ratio is 3: 1-5: the mixed solution of 1 acetonitrile and the dilution heat of sulfuric acid of 0.5mol/L, elution time are 12h.
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Application publication date: 20120328 |