CN109254045B - Cobalt-based nitride sensor for detecting praziquantel and preparation method thereof - Google Patents

Cobalt-based nitride sensor for detecting praziquantel and preparation method thereof Download PDF

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CN109254045B
CN109254045B CN201811306513.XA CN201811306513A CN109254045B CN 109254045 B CN109254045 B CN 109254045B CN 201811306513 A CN201811306513 A CN 201811306513A CN 109254045 B CN109254045 B CN 109254045B
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praziquantel
cobalt
electrode
template
molecularly imprinted
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CN109254045A (en
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刘召壹
张勇
杜斌
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University of Jinan
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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
    • 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
    • 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
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
    • G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles

Abstract

The invention discloses a preparation method of a cobalt-based nitride sensor for detecting praziquantel. Belongs to the technical field of novel nanometer functional materials and chemical biosensors. According to the method, firstly, a cobalt nitride nanosheet array is prepared on a disposable throwable electrode, a polydopamine film containing an electron mediator and a molecularly imprinted polymer taking praziquantel molecules as template molecules are sequentially and directly prepared on the cobalt nitride nanosheet array in sequence by utilizing the large specific surface area and the high adsorption activity to amino groups through an in-situ growth method, and after the template molecules are eluted, the original positions of the template molecules are changed into cavities, namely the molecularly imprinted polymer of the template molecules is eluted, so that the cobalt-based nitride sensor for detecting the praziquantel is prepared.

Description

Cobalt-based nitride sensor for detecting praziquantel and preparation method thereof
Technical Field
The invention relates to a preparation method and application of an electrochemical sensor. Belongs to the technical field of novel nanometer functional materials and chemical biosensors.
Background
Praziquantel (Praziquantel) is an anthelmintic used in humans and animals, and is used exclusively for the treatment of tapeworms and trematodes. However, due to its high toxicity, the medication must be used strictly as required. Otherwise, dizziness, headache, nausea, abdominal pain, diarrhea, asthenia, soreness of limbs, etc. may be caused, and palpitation, chest distress, etc. may occur in severe cases, which may occasionally induce mental disorder or gastrointestinal hemorrhage. Animals need to be slaughtered and marketed in the drug withdrawal period of more than 5-7 days. When people eat animal food with residual anthelmintic, the drug toxicity can be transferred to human beings, thereby harming human health. At present, methods for detecting praziquantel molecules mainly comprise an enzyme-linked immunosorbent assay, a high performance liquid chromatography, a mass spectrometry method and the like. The method is expensive and complex in operation, and the detection can be carried out only after professional training is required for a laboratory worker. Therefore, the method for quickly and selectively detecting the praziquantel is very important to public health and has wide market application prospect.
The molecular imprinting electrochemical sensor has high specificity selectivity, excellent stability, excellent reproducibility, wide detection range and bottom detection limit. The sensor has the advantages of simple preparation, convenient detection, high sensitivity, low cost and the like, and can be widely applied to the fields of chromatographic separation, membrane separation, solid-phase extraction, drug controlled release, chemical sensing and the like. Molecularly Imprinted Polymers (MIPs), also known as "plastic antibodies", are capable of specifically recognizing and selectively adsorbing a specific target molecule (i.e., template molecule). The molecular imprinting technology has many advantages, such as corrosion resistance of organic reagents, good stability, high temperature resistance and simple preparation. Thus, MIP electrochemical sensors based on MIP in combination with electrochemical sensors (MIP-ECS) have attracted a hot interest in the field of electroanalytical chemistry, especially the detection of small molecule contaminants, over the last few years. However, in the preparation process of the traditional MIP-ECS, the defects of difficult elution of template molecules, difficult control of the thickness of the imprinted membrane, poor reproducibility and the like exist, and the application of the molecularly imprinted membrane in an electrochemical sensor is limited. The problems, especially the technical problems that the sensitivity of the electrochemical sensor is reduced due to the fact that the thickness of the molecularly imprinted membrane is not easy to control, and the stability and the reproducibility are reduced due to the fact that the molecularly imprinted membrane is easy to fall off from the surface of an electrode in the elution process, limit the application of MIP _ ECS, so that the research of a new molecularly imprinted polymer synthesis method, a new molecularly imprinted membrane electrode modification method and a method for combining the molecularly imprinted membrane and a substrate material for solving the preparation and application problems of MIP-ECS has important research significance and market value.
Disclosure of Invention
The invention aims to provide a preparation method of a cobalt-based nitride sensor for detecting praziquantel, which has the advantages of strong specificity, simple preparation, convenient detection, high sensitivity and low cost. Based on the purpose, firstly, a cobalt nitride nanosheet array is prepared on a disposable throwable electrode, and by utilizing the large specific surface area and the high adsorption activity to amino groups of the cobalt nitride nanosheet array, a polydopamine film containing an electron mediator and a molecularly imprinted polymer taking a praziquantel molecule as a template molecule are sequentially and directly prepared on the cobalt nitride nanosheet array by an in-situ growth method, and after the template molecule is eluted, the original position of the template molecule is changed into a cavity, namely the molecularly imprinted polymer of the template molecule is eluted, so that the cobalt-based nitride sensor for detecting the praziquantel is prepared. When the detection device is used for detecting praziquantel molecules, the cobalt-based nitride sensor for detecting praziquantel is inserted into a solution to be detected, and the praziquantel molecules in the solution to be detected are adsorbed into cavities of NIP. The higher the concentration of the praziquantel molecules in the solution to be tested is, the more praziquantel molecules are adsorbed into the cavities of the NIP. When electrochemical detection is carried out, the intensity of detection current is reduced along with the increase of praziquantel molecules adsorbed in the holes of the NIP, so that the concentration of the praziquantel molecules in the solution to be detected can be qualitatively and quantitatively determined according to the reduction degree of the current intensity.
The technical scheme adopted by the invention is as follows:
1. the preparation method of the cobalt-based nitride sensor for detecting the praziquantel is characterized in that the cobalt-based nitride sensor for detecting the praziquantel is obtained by growing a template-free molecularly imprinted polymer NIP in situ on a cobalt nitride nanosheet array electrode CoN-nanoarray; the template-free molecularly imprinted polymer NIP is a molecularly imprinted polymer without template molecules; the molecularly imprinted polymer without the template molecule is obtained by eluting the template molecule from a MIP containing the template molecularly imprinted polymer; the MIP containing the template molecule engram polymer is the MIP containing the template molecule; the template molecule is a praziquantel molecule;
2. the preparation method of the cobalt nitride nanosheet array electrode CoN-nanoarray in the technical scheme 1 comprises the following preparation steps:
(1) carrying out ultrasonic cleaning treatment on the disposable throwable electrode by respectively using dilute hydrochloric acid, absolute ethyl alcohol and deionized water so as to remove an oxide layer and surface impurities of the disposable throwable electrode;
(2) weighing 1-3 mmol of cobalt nitrate hexahydrate Co (NO3)2·6H2O and 3-9 mmol of urea CO (NH)2)2Placing the mixture into a 50mL beaker, adding 30mL deionized water, stirring until the mixture is clear, and then transferring the mixture into a 50mL polytetrafluoroethylene reaction kettle;
(3) putting the disposable throwable electrode processed in the step (1) into the solution in the reaction kettle in the step (2), and reacting at the temperature of 100-130 ℃ for 9-12 hours to prepare a cobalt hydroxide nanosheet array precursor electrode;
(4) inserting the cobalt hydroxide nanosheet array precursor electrode obtained in the step (3) into ammonia water for 5-30 seconds, taking out, heating to 340-400 ℃ in an ammonia environment, keeping for 4-8 hours, then naturally cooling to room temperature in the ammonia environment, then inserting the cobalt hydroxide nanosheet array precursor electrode into phosphate buffer solution PBS containing dopamine, ammonium persulfate and cobalt nitrate, reacting for 4-6 hours at the temperature of 20-40 ℃, taking out, and performing immersion washing for 2-4 times by using deionized water to prepare a cobalt nitride nanosheet array electrode CoN-nanoarray;
the disposable and disposable electrode is selected from one of the following electrodes: foam iron, foam copper, pure iron sheets, pure copper sheets, pure iron sheets, pure silicon wafers and conductive carbon cloth;
in the phosphate buffer solution PBS containing dopamine, ammonium persulfate and cobalt nitrate: the concentration of dopamine is 2-5 mg/mL, the concentration of ammonium persulfate is 3-8 mg/mL, the concentration of cobalt nitrate is 0.1-0.5 mg/mL, the concentration of phosphate buffer solution PBS is 0.1mol/L, and the pH value is 7.2-8.5;
3. the preparation method of the MIP containing the template molecularly imprinted polymer grown in situ by CoN-nanoarray in the technical scheme 1 comprises the following preparation steps:
(1) respectively weighing 0.25-0.45 mmol of template molecules and 3-5 mmol of 2-methacrylic acid MAA in an ampoule bottle, adding 8-15 mL of acetonitrile, and carrying out ultrasonic treatment for 30min until all the template molecules and the 2-methacrylic acid MAA are dissolved;
(2) adding 15-25 mmol of ethylene glycol dimethacrylate EDMA into the solution obtained in the step (1), and carrying out ultrasonic treatment for 30min until the mixture is uniformly mixed to obtain a precursor mixed solution;
(3) the CoN-nanoarray clip prepared in the technical scheme 2Inserting the precursor mixed solution in the step (2) on a rotary stirrer, and adding the precursor mixed solution into a reactor to obtain a solution with a certain volume ratio N2Under the temperature of environment and water bath of 20-40 ℃, rotationally stirring at the speed of 5-200 r/s, simultaneously dripping 1mmol of azobisisobutyronitrile AIBN into the mixed solution at the speed of 1-20 drops/s to initiate polymerization, and obtaining the in-situ grown MIP containing the template molecularly imprinted polymer on the CoN-nanoarray;
4. the preparation steps of the template-free molecularly imprinted polymer NIP for in-situ CoN-nanoarray growth in the technical scheme 1 are as follows: immersing the MIP which is obtained in the technical scheme 3 and grows in situ on the CoN-nanoarray and contains the template molecularly imprinted polymer into an eluant, eluting the template molecule for 5-20 min at room temperature, and then taking out to obtain the NIP without the template molecularly imprinted polymer; the eluent is a mixed solution of formic acid and methanol, wherein the volume ratio of the formic acid to the methanol is 9 (1-5);
5. the preparation steps of the cobalt-based nitride sensor for detecting praziquantel in the technical scheme 1 are as follows: washing the template-free molecularly imprinted polymer NIP which grows in situ on the CoN-nanoarray prepared in the technical scheme 2-4 with deionized water for 2-4 times, and airing at room temperature to prepare the cobalt-based nitride sensor for detecting the praziquantel;
6. the cobalt-based nitride sensor for detecting praziquantel prepared by the technical scheme 1-5 is applied to detection of praziquantel molecules, and comprises the following application steps:
(1) preparing a standard solution: preparing a group of praziquantel molecule standard solutions with different concentrations including blank standard samples;
(2) modification of a working electrode: inserting a cobalt-based nitride sensor for detecting praziquantel as a working electrode into the praziquantel molecule standard solutions with different concentrations prepared in the step (1), incubating for 10min, taking out, and washing with deionized water for 3 times;
(3) drawing a working curve: taking a saturated calomel electrode as a reference electrode, taking a platinum wire electrode as a counter electrode, forming a three-electrode system with the modified working electrode in the step (2), connecting the three-electrode system with an electrochemical workstation, and sequentially adding the three-electrode system into an electrolytic cell15mL phosphate buffer PBS; detecting a current response of the assembled working electrode by Differential Pulse Voltammetry (DPV); the response current intensity of the blank sample is recordedI 0The response current intensity of the standard solution containing praziquantel molecules at different concentrations is recordedI iThe difference in response to the decrease in current intensity is ΔI = I 0-I i，ΔIAnd mass concentration of praziquantel molecule standard solutionCWith a linear relationship therebetween, plotting ΔI－CA working curve; the concentration of the phosphate buffer solution PBS is 10mmol/L, pH, and the value is 7.4; the parameters during DPV detection are set as follows: the range and the direction are 0-1V, the step is 0.05V, the pulse time is 0.05s, the sampling time is 0.016s, and the pulse period is 0.5 s;
(4) detection of praziquantel molecules in a sample to be detected: replacing the praziquantel molecule standard solution in the step (1) with a sample to be detected, detecting according to the methods in the steps (2) and (3), and responding to the difference delta of the reduction of the current intensityIAnd working curve to obtain the content of praziquantel molecules in the sample to be detected.
Advantageous results of the invention
(1) The cobalt-based nitride sensor for detecting praziquantel is simple to prepare, convenient to operate, low in cost, applicable to portable detection and has market development prospect, and the sample can be quickly, sensitively and selectively detected;
(2) according to the invention, the molecularly imprinted polymer is grown in situ on the cobalt nitride nanosheet array electrode CoN-nanoarray for the first time, on one hand, more and more uniform molecularly imprinted polymers can be grown by utilizing the large specific surface area of the CoN-nanoarray, and the CoN-nanoarray has excellent electron transfer capacity, so that the detection sensitivity is greatly improved; on the other hand, when dopamine is polymerized onto a cobalt nitride nanosheet array in situ, cobalt ions are creatively doped as an electronic mediator, and electrochemical response current is directly generated during detection, so that the sensor can directly detect in a buffer solution without adding other mediator substances, thereby further reducing the signal background, improving the detection sensitivity, greatly reducing the detection cost and reducing the environmental pollution;
(3) according to the invention, the high adsorption activity of the nitride on amino and the large specific surface area of the nano array electrode are combined with dopamine, so that when dopamine is polymerized in situ on the surface of the cobalt nitride nanosheet array, a sufficiently thin polydopamine film is formed and simultaneously the polydopamine film is uniformly covered on the cobalt nitride nanosheet array, thereby laying a better foundation for the next step of polymerizing the molecularly imprinted polymer; then utilizing strong adsorption and connection effects of polydopamine on amino groups rich in the molecularly imprinted polymer, skillfully using CoN-nanoarray as a stirrer, immersing and stirring the polydopamine in a molecularly imprinted precursor mixed solution, and directly growing the molecularly imprinted polymer with the film thickness in situ on the surface of the CoN-nanoarray by controlling the stirring speed, the dropping speed of a polymerization reaction initiator and the polymerization reaction temperature, so that the CoN-nanoarray can firmly load the molecularly imprinted polymer on one hand, and the stability and the reproducibility of the prepared electrochemical sensor are obviously improved; on the other hand, the film forming thickness of the molecularly imprinted polymer on the surface of the electrode can be effectively controlled, and the technical problem of poor reproducibility caused by the fact that the film forming thickness of the molecularly imprinted film on the surface of the electrode cannot be controlled is solved; in addition, the preparation method of the invention has important scientific significance and application value for effectively controlling the thickness of the formed film and coating the electronic mediator in situ, and fully improving the sensitivity and detection limit of the electrochemical sensor based on the molecular imprinting.
Detailed Description
EXAMPLE 1 preparation of CoN-nanoarray
(1) Carrying out ultrasonic cleaning treatment on the disposable throwable electrode by respectively using dilute hydrochloric acid, absolute ethyl alcohol and deionized water so as to remove an oxide layer and surface impurities of the disposable throwable electrode;
(2) 1mmol of cobalt nitrate hexahydrate Co (NO) was weighed3)2·6H2O and 3mmol Urea CO (NH)2)2Placing the mixture into a 50mL beaker, adding 30mL deionized water, stirring until the mixture is clear, and then transferring the mixture into a 50mL polytetrafluoroethylene reaction kettle;
(3) putting the disposable throwable electrode processed in the step (1) into the solution in the reaction kettle in the step (2), and reacting at the temperature of 100 ℃ for 12 hours to prepare a cobalt hydroxide nanosheet array precursor electrode;
(4) inserting the cobalt hydroxide nanosheet array precursor electrode obtained in the step (3) into ammonia water for 5 seconds, then taking out, heating to 340 ℃ in an ammonia environment, keeping for 8 hours, then naturally cooling to room temperature in the ammonia environment, then inserting the cobalt hydroxide nanosheet array precursor electrode into phosphate buffer solution PBS containing dopamine, ammonium persulfate and cobalt nitrate, reacting for 4 hours at the temperature of 20 ℃, taking out, and performing immersion washing for 2 times by using deionized water to prepare a cobalt nitride nanosheet array electrode CoN-nanoarray;
wherein the disposable throwable electrode is foam iron; the concentration of dopamine is 2 mg/mL, the concentration of ammonium persulfate is 3 mg/mL, the concentration of cobalt nitrate is 0.1 mg/mL, the concentration of phosphate buffer solution PBS is 0.1mol/L, and the pH value is 7.2.
EXAMPLE 2 preparation of CoN-nanoarray
(1) Carrying out ultrasonic cleaning treatment on the disposable throwable electrode by respectively using dilute hydrochloric acid, absolute ethyl alcohol and deionized water so as to remove an oxide layer and surface impurities of the disposable throwable electrode;
(2) weighing 2 mmol cobalt nitrate hexahydrate Co (NO)3)2·6H2O and 6 mmol Urea CO (NH)2)2Placing the mixture into a 50mL beaker, adding 30mL deionized water, stirring until the mixture is clear, and then transferring the mixture into a 50mL polytetrafluoroethylene reaction kettle;
(3) putting the disposable throwable electrode processed in the step (1) into the solution in the reaction kettle in the step (2), and reacting for 11 hours at the temperature of 110 ℃ to prepare a cobalt hydroxide nanosheet array precursor electrode;
(4) inserting the cobalt hydroxide nanosheet array precursor electrode obtained in the step (3) into ammonia water for 15 seconds, then taking out, heating to 370 ℃ in an ammonia environment, keeping for 6 hours, then continuing to naturally cool to room temperature in the ammonia environment, then inserting the cobalt hydroxide nanosheet array precursor electrode into phosphate buffer solution PBS containing dopamine, ammonium persulfate and cobalt nitrate, reacting for 5 hours at the temperature of 30 ℃, taking out, and performing immersion washing for 3 times by using deionized water to prepare a cobalt nitride nanosheet array electrode CoN-nanoarray;
wherein the disposable throwable electrode is a pure copper sheet; the concentration of dopamine is 3.5 mg/mL, the concentration of ammonium persulfate is 6.2 mg/mL, the concentration of cobalt nitrate is 0.3 mg/mL, the concentration of phosphate buffer solution PBS is 0.1mol/L, and the pH value is 8.0.
EXAMPLE 3 preparation of CoN-nanoarray
(1) Carrying out ultrasonic cleaning treatment on the disposable throwable electrode by respectively using dilute hydrochloric acid, absolute ethyl alcohol and deionized water so as to remove an oxide layer and surface impurities of the disposable throwable electrode;
(2) weighing 3mmol cobalt nitrate hexahydrate Co (NO)3)2·6H2O and 9mmol Urea CO (NH)2)2Placing the mixture into a 50mL beaker, adding 30mL deionized water, stirring until the mixture is clear, and then transferring the mixture into a 50mL polytetrafluoroethylene reaction kettle;
(3) putting the disposable throwable electrode processed in the step (1) into the solution in the reaction kettle in the step (2), and reacting at the temperature of 130 ℃ for 9 hours to prepare a cobalt hydroxide nanosheet array precursor electrode;
(4) inserting the cobalt hydroxide nanosheet array precursor electrode obtained in the step (3) into ammonia water for 30 seconds, then taking out, heating to 400 ℃ in an ammonia environment, keeping for 4 hours, then continuing to naturally cool to room temperature in the ammonia environment, then inserting the cobalt hydroxide nanosheet array precursor electrode into phosphate buffer solution PBS containing dopamine, ammonium persulfate and cobalt nitrate, reacting at the temperature of 40 ℃ for 6 hours, taking out, and performing immersion washing with deionized water for 4 times to prepare a cobalt nitride nanosheet array electrode CoN-nanoarray;
wherein the disposable throwable electrode is a conductive carbon cloth; the concentration of dopamine is 5mg/mL, the concentration of ammonium persulfate is 8mg/mL, the concentration of cobalt nitrate is 0.5mg/mL, the concentration of phosphate buffer solution PBS is 0.1mol/L, and the pH value is 8.5.
Embodiment 4 preparation method of cobalt-based nitride sensor for detecting praziquantel
(1) Respectively weighing 0.25 mmol of template molecules and 3mmol of 2-methacrylic acid MAA in an ampoule bottle, adding 8 mL of acetonitrile, and performing ultrasonic treatment for 30min until all the template molecules and the 3mmol of 2-methacrylic acid MAA are dissolved;
(2) adding 15 mmol of ethylene glycol dimethacrylate EDMA into the solution obtained in the step (1), and carrying out ultrasonic treatment for 30min until the mixture is uniformly mixed to obtain a precursor mixed solution;
(3) CoN-nanoarray prepared in example 1 was clamped to a rotary stirrer and inserted into the precursor mixed solution in step (2) in N2Under the temperature of environment and water bath 20 ℃, stirring in a rotating way at the speed of 200 r/s, simultaneously dripping 1mmol of azobisisobutyronitrile AIBN into the mixed solution at the speed of 1 d/s to initiate polymerization, and obtaining the in-situ grown MIP containing the template molecularly imprinted polymer on the CoN-nanoarray;
(4) immersing the MIP which is obtained in the step (3) and grows in situ on the CoN-nanoarray and contains the template molecularly imprinted polymer into an eluant, eluting the template molecule for 5 min at room temperature, and then taking out to obtain the NIP without the template molecularly imprinted polymer; continuously washing with deionized water for 2 times, and airing at room temperature to obtain the cobalt-based nitride sensor for detecting praziquantel;
the eluent is a mixed solution of formic acid and methanol, wherein the volume ratio of the formic acid to the methanol is 9: 1.
Example 5 preparation method of cobalt-based nitride sensor for detecting praziquantel
(1) Respectively weighing 0.35mmol of template molecules and 4 mmol of 2-methacrylic acid MAA in an ampoule bottle, adding 12 mL of acetonitrile, and carrying out ultrasonic treatment for 30min until all the template molecules and the 2-methacrylic acid MAA are dissolved;
(2) adding 18 mmol of ethylene glycol dimethacrylate EDMA into the solution obtained in the step (1), and carrying out ultrasonic treatment for 30min until the mixture is uniformly mixed to obtain a precursor mixed solution;
(3) clamping the CoN-nanoarray prepared in the technical scheme 2 on a rotary stirrer, inserting the CoN-nanoarray into the precursor mixed solution in the step (2), and adding N2Stirring at 60 rpm while dropping 10 drops/s in the mixed solution at 30 deg.C in water bath1mmol of azobisisobutyronitrile AIBN is initiated to polymerize, and the in-situ grown MIP containing the template molecularly imprinted polymer is obtained on the CoN-nanoarray;
(4) immersing the MIP which is obtained in the step (3) and grows in situ on the CoN-nanoarray and contains the template molecularly imprinted polymer into an eluant, eluting the template molecule for 10min at room temperature, and then taking out to obtain the NIP without the template molecularly imprinted polymer; continuously washing with deionized water for 3 times, and airing at room temperature to obtain the cobalt-based nitride sensor for detecting praziquantel;
the eluent is a mixed solution of formic acid and methanol, wherein the volume ratio of the formic acid to the methanol is 9: 3.
Example 6 preparation method of cobalt-based nitride sensor for detecting praziquantel
(1) Respectively weighing 0.45mmol of template molecules and 5mmol of 2-methacrylic acid MAA in an ampoule bottle, adding 15mL of acetonitrile, and carrying out ultrasonic treatment for 30min until all the template molecules and the 5mmol of 2-methacrylic acid MAA are dissolved;
(2) adding 25mmol of ethylene glycol dimethacrylate EDMA into the solution obtained in the step (1), and carrying out ultrasonic treatment for 30min until the mixture is uniformly mixed to obtain a precursor mixed solution;
(3) clamping the CoN-nanoarray prepared in the technical scheme 2 on a rotary stirrer, inserting the CoN-nanoarray into the precursor mixed solution in the step (2), and adding N2Under the temperature of environment and water bath 40 ℃, rotationally stirring at the speed of 5 r/s, simultaneously dripping 1mmol of azobisisobutyronitrile AIBN into the mixed solution at the speed of 20 drops/s to initiate polymerization, and obtaining the in-situ grown MIP containing the template molecularly imprinted polymer on the CoN-nanoarray;
(4) immersing the MIP which is obtained in the step (3) and grows in situ on the CoN-nanoarray and contains the template molecularly imprinted polymer into an eluant, eluting the template molecule for 20min at room temperature, and then taking out to obtain the NIP without the template molecularly imprinted polymer; continuously washing with deionized water for 4 times, and airing at room temperature to obtain the cobalt-based nitride sensor for detecting praziquantel;
the eluent is a mixed solution of formic acid and methanol, wherein the volume ratio of the formic acid to the methanol is 9: 5.
Example 7 the cobalt-based nitride sensor for detecting praziquantel prepared in examples 1 to 6 is applied to detection of praziquantel molecules, and comprises the following steps:
(1) preparing a standard solution: preparing a group of praziquantel molecule standard solutions with different concentrations including blank standard samples;
(2) modification of a working electrode: inserting a cobalt-based nitride sensor for detecting praziquantel as a working electrode into the praziquantel standard solutions with different concentrations prepared in the step (1), incubating for 10min, taking out, and washing with deionized water for 3 times;
(3) drawing a working curve: taking a saturated calomel electrode as a reference electrode, taking a platinum wire electrode as a counter electrode, forming a three-electrode system with the modified working electrode in the step (2), connecting the three-electrode system with an electrochemical workstation, and sequentially adding 15mL PBS into an electrolytic bath; detecting a current response of the assembled working electrode by Differential Pulse Voltammetry (DPV); the response current intensity of the blank sample is recordedI 0The response current intensity of the standard solution containing praziquantel molecules at different concentrations is recordedI iThe difference in response to the decrease in current intensity is ΔI = I 0-I i，ΔIAnd mass concentration of praziquantel standard solutionCWith a linear relationship therebetween, plotting ΔI－CA working curve; the PBS is 10mmol/L phosphate buffer solution, and the pH value of the phosphate buffer solution is 7.4; the parameters during DPV detection are set as follows: the range and the direction are 0-1V, the step is 0.05V, the pulse time is 0.05s, the sampling time is 0.016s, and the pulse period is 0.5 s;
(4) detection of praziquantel in a sample to be detected: replacing the praziquantel standard solution in the step (1) with a sample to be detected, detecting according to the methods in the steps (2) and (3), and responding to the difference delta of the reduction of the current intensityIAnd working curve to obtain the content of the praziquantel in the sample to be detected.
Example 8 the cobalt-based nitride sensor for detecting praziquantel prepared in examples 1 to 6 was applied to the detection of praziquantel according to the detection procedure of example 7, and had a linear range of 0.0001 to 100 mmol/L and a detection limit of 30 nmol/L.
Example 9 detection of Praziquantel in pig urine samples
Accurately transferring a swine urine sample, adding a praziquantel standard solution with a certain mass concentration, taking a swine urine sample without praziquantel as a blank, carrying out a standard recovery experiment, detecting by using the cobalt-based nitride sensor for detecting praziquantel prepared in examples 1-6 according to the steps of example 7, and determining the recovery rate of praziquantel in the swine urine sample, wherein the detection results are shown in table 1:
TABLE 1 detection results of Praziquantel in pig urine samples
Figure 208599DEST_PATH_IMAGE002
The detection results in the table 1 show that the Relative Standard Deviation (RSD) of the results is less than 3.2%, the average recovery rate is 98.0-101.6%, and the method can be used for detecting praziquantel in pig urine, and is high in sensitivity, strong in specificity, and accurate and reliable in result.
EXAMPLE 10 detection of Praziquantel in sheep urine samples
Accurately transferring a certain amount of sheep urine samples, adding a praziquantel standard solution with a certain mass concentration, taking the sheep urine samples without praziquantel as blanks, carrying out a labeling recovery experiment, detecting the recovery rate of praziquantel in the sheep urine samples by using the cobalt-based nitride sensor for detecting praziquantel prepared in examples 1-6 according to the steps of example 7, and determining the detection results shown in table 2:
TABLE 2 detection results of Praziquantel in sheep urine samples
Figure 125740DEST_PATH_IMAGE004
The detection results in the table 2 show that the Relative Standard Deviation (RSD) of the results is less than 3.2%, the average recovery rate is 99.0-101%, and the method can be used for detecting praziquantel in sheep urine, and is high in sensitivity, strong in specificity, and accurate and reliable in result.

Claims (3)

1. The preparation method of the cobalt-based nitride sensor for detecting the praziquantel is characterized in that the cobalt-based nitride sensor for detecting the praziquantel is obtained by growing a template-free molecularly imprinted polymer NIP in situ on a cobalt nitride nanosheet array CoN-nanoarray; the template-free molecularly imprinted polymer NIP is a molecularly imprinted polymer without template molecules; the molecularly imprinted polymer without the template molecule is obtained by eluting the template molecule from a MIP containing the template molecularly imprinted polymer; the MIP containing the template molecule engram polymer is the MIP containing the template molecule; the template molecule is praziquantel molecule, MIP containing template molecule imprinted polymer is directly grown on CoN-nanoarray in situ, and the preparation method comprises the following preparation steps:
(1) respectively weighing 0.25-0.45 mmol of template molecules and 3-5 mmol of 2-methacrylic acid MAA in an ampoule bottle, adding 8-15 mL of acetonitrile, and carrying out ultrasonic treatment for 30min until all the template molecules and the 2-methacrylic acid MAA are dissolved;
(2) adding 15-25 mmol of ethylene glycol dimethacrylate EDMA into the solution obtained in the step (1), and carrying out ultrasonic treatment for 30min until the mixture is uniformly mixed to obtain a precursor mixed solution;
(3) clamping CoN-nanoarray on a rotary stirrer, and inserting the CoN-nanoarray into the precursor mixed solution in the step (2), wherein N is2Under the temperature of environment and water bath of 20-40 ℃, rotationally stirring at the speed of 5-200 r/s, simultaneously dripping 1mmol of azobisisobutyronitrile AIBN into the mixed solution at the speed of 1-20 drops/s to initiate polymerization, and obtaining the in-situ grown MIP containing the template molecularly imprinted polymer on the CoN-nanoarray;
the preparation method of the template-free molecularly imprinted polymer NIP comprises the following steps: immersing the obtained MIP which grows in situ on the CoN-nanoarray and contains the template molecularly imprinted polymer in an eluant, eluting the template molecule for 5-20 min at room temperature, and then taking out to obtain the NIP without the template molecularly imprinted polymer; the eluent is a mixed solution of formic acid and methanol, wherein the volume ratio of the formic acid to the methanol is 9 (1-5);
the preparation steps of the sensor are as follows: and (3) washing the prepared template-free molecularly imprinted polymer NIP growing in situ on the CoN-nanoarray with deionized water for 2-4 times, and airing at room temperature to obtain the cobalt-based nitride sensor for detecting the praziquantel.
2. The preparation method of a cobalt-based nitride sensor for detecting praziquantel according to claim 1, wherein the preparation method of the CoN-nanoarray comprises the following preparation steps:
(1) carrying out ultrasonic cleaning treatment on the disposable throwable electrode by respectively using dilute hydrochloric acid, absolute ethyl alcohol and deionized water so as to remove an oxide layer and surface impurities of the disposable throwable electrode;
(2) weighing 1-3 mmol cobalt nitrate hexahydrate Co (NO)3)2·6H2O and 3-9 mmol of urea CO (NH)2)2Placing the mixture into a 50mL beaker, adding 30mL deionized water, stirring until the mixture is clear, and then transferring the mixture into a 50mL polytetrafluoroethylene reaction kettle;
(3) putting the disposable throwable electrode processed in the step (1) into the solution in the reaction kettle in the step (2), and reacting at the temperature of 100-130 ℃ for 9-12 hours to prepare a cobalt hydroxide nanosheet array precursor electrode;
(4) inserting the cobalt hydroxide nanosheet array precursor electrode obtained in the step (3) into ammonia water for 5-30 seconds, taking out, heating to 340-400 ℃ in an ammonia environment, keeping for 4-8 hours, then naturally cooling to room temperature in the ammonia environment, then inserting the cobalt hydroxide nanosheet array precursor electrode into phosphate buffer solution PBS containing dopamine, ammonium persulfate and cobalt nitrate, reacting for 4-6 hours at the temperature of 20-40 ℃, taking out, and performing immersion washing for 2-4 times by using deionized water to prepare a cobalt nitride nanosheet array CoN-nanoarray;
the disposable and disposable electrode is selected from one of the following electrodes: foam nickel, foam copper, pure nickel sheets, pure copper sheets, pure iron sheets, pure silicon sheets and conductive carbon cloth;
in the phosphate buffer solution PBS containing dopamine, ammonium persulfate and cobalt nitrate: the concentration of dopamine is 2-5 mg/mL, the concentration of ammonium persulfate is 3-8 mg/mL, the concentration of cobalt nitrate is 0.1-0.5 mg/mL, the concentration of phosphate buffer solution PBS is 0.1mol/L, and the pH value is 7.2-8.5.
3. The cobalt-based nitride sensor for detecting praziquantel prepared by the preparation method of any one of claims 1 to 2 is applied to detection of praziquantel, and the detection steps are as follows:
(1) preparing a standard solution: preparing a group of praziquantel standard solutions with different concentrations including blank standard samples;
(2) modification of a working electrode: taking a cobalt-based nitride sensor for detecting praziquantel as a working electrode, inserting the cobalt-based nitride sensor into the praziquantel standard solutions with different concentrations prepared in the step (1), incubating for 10min, taking out, and washing with deionized water for 3 times;
(3) drawing a working curve: taking a saturated calomel electrode as a reference electrode, taking a platinum wire electrode as a counter electrode, forming a three-electrode system with the modified working electrode in the step (2), connecting the three-electrode system with an electrochemical workstation, and sequentially adding 15mL phosphate buffer solution PBS into an electrolytic bath; detecting a current response of the assembled working electrode by Differential Pulse Voltammetry (DPV); the response current intensity of the blank sample is recorded as I0The response current intensity of the standard solution containing praziquantel with different concentrations is recorded as IiThe difference of response current intensity is Δ I ═ I0-IiThe mass concentration C of the praziquantel standard solution is in a linear relation with the mass concentration I of the praziquantel standard solution, and a working curve of the mass concentration C is drawn; the concentration of the phosphate buffer solution PBS is 10mmol/L, and the pH value is 7.4; the parameters during DPV detection are set as follows: the range and the direction are 0-1V, the step is 0.05V, the pulse time is 0.05s, the sampling time is 0.016s, and the pulse period is 0.5 s;
(4) detection of praziquantel in a sample to be detected: and (3) replacing the praziquantel standard solution in the step (1) with a sample to be detected, detecting according to the methods in the steps (2) and (3), and obtaining the content of the praziquantel in the sample to be detected according to the difference value delta I of the reduction of the response current intensity and the working curve.
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102901754A (en) * 2011-07-27 2013-01-30 中国科学院电子学研究所 Electropolymerization molecular imprinting technology-based double-parameter composite micro-sensor and preparation thereof
CN105738437A (en) * 2016-02-25 2016-07-06 济南大学 Preparing method and application of electrochemistry parathion sensor based on metal and metal oxide co-doped nanometer composite
CN108519365A (en) * 2018-04-08 2018-09-11 上海应用技术大学 Surface enhanced Raman spectroscopy sensor and preparation method thereof based on electrochemical deposition and molecular engram

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102901754A (en) * 2011-07-27 2013-01-30 中国科学院电子学研究所 Electropolymerization molecular imprinting technology-based double-parameter composite micro-sensor and preparation thereof
CN105738437A (en) * 2016-02-25 2016-07-06 济南大学 Preparing method and application of electrochemistry parathion sensor based on metal and metal oxide co-doped nanometer composite
CN108519365A (en) * 2018-04-08 2018-09-11 上海应用技术大学 Surface enhanced Raman spectroscopy sensor and preparation method thereof based on electrochemical deposition and molecular engram

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Molecularly imprinted polymeric nanoparticles decorated with Au NPs for highly sensitive and selective glucose detection;Wei Zhao 等;《Biosensors and Bioelectronics》;20170915;第100卷;第497-503页 *
基于分子印迹技术的电化学发光分析;杨钰昆 等;《化学进展》;20160915;第28卷(第9期);第1351-01362页 *
电化学发光-分子印迹传感器的制备及其在海洛因检测中的应用;商哲一 等;《分析化学(FENXI HUAXUE) 研究简报》;20140630;第42卷(第6期);第904-908页 *

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