CN109254044B - Preparation method and application of macrolide antibiotic sensor based on FeN - Google Patents
Preparation method and application of macrolide antibiotic sensor based on FeN Download PDFInfo
- Publication number
- CN109254044B CN109254044B CN201811306512.5A CN201811306512A CN109254044B CN 109254044 B CN109254044 B CN 109254044B CN 201811306512 A CN201811306512 A CN 201811306512A CN 109254044 B CN109254044 B CN 109254044B
- Authority
- CN
- China
- Prior art keywords
- electrode
- template
- macrolide antibiotic
- iron nitride
- fen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Classifications
-
- 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
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Analytical Chemistry (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Electrochemistry (AREA)
- Engineering & Computer Science (AREA)
- Nanotechnology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Plasma & Fusion (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a preparation method of a macrolide antibiotic sensor based on iron nitride. Belongs to the technical field of novel nanometer functional materials and chemical biosensors. According to the invention, firstly, an iron 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 iron nitride nanosheet array, a polydopamine film and a molecular imprinting polymer which is coated with luminol in situ and takes macrolide antibiotic molecules as template molecules are sequentially and directly prepared on the iron nitride nanosheet array by adopting 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 molecular imprinting polymer of the template molecules is eluted, so that the preparation of the iron nitride-based macrolide antibiotic sensor is completed.
Description
Technical Field
The invention relates to a preparation method and application of an electrochemiluminescence sensor. Belongs to the technical field of novel nanometer functional materials and chemical biosensors.
Background
Macrolide Antibiotics (MA) are a generic name of antibacterial drugs having a 12-16 carbon lactone ring in a molecular structure, inhibit bacterial protein synthesis by blocking the activity of peptidyl transferase in 50s ribosome, and belong to rapid bacteriostats. Mainly used for treating infection of aerobic gram positive coccus and negative coccus, certain anaerobe, legionella, mycoplasma, chlamydia and the like. When people eat animal food with residual antibiotics, drug resistance generated by animals can be transferred to human beings, thereby harming human health. At present, methods for detecting macrolide antibiotic molecules mainly comprise an enzyme-linked immunosorbent assay, a high performance liquid chromatography, a mass spectrometry 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 developing the macrolide antibiotics with high selectivity and sensitivity is very important to public health and has wide market application prospect.
The electroanalytical chemical sensors include electrochemical sensors, electrochemiluminescence sensors, photoelectrochemical sensors and the like, and the sensors have 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 electroanalytical chemical sensors (MIP-ECS) based on the combination of MIPs with electroanalytical chemical sensors have attracted a great deal of interest in the field of electroanalytical chemistry, particularly 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 electroanalytical chemical 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 macrolide antibiotic sensor based on iron nitride, which has the advantages of strong specificity, simple preparation, convenient detection, high sensitivity and low cost. Based on the purpose, firstly, an iron nitride nanosheet array is prepared on a disposable throwable electrode, a polydopamine film and a molecular imprinting polymer which is coated with luminol in situ and takes macrolide antibiotic molecules as template molecules are sequentially and directly prepared on the iron nitride nanosheet array by utilizing the large specific surface area and the high adsorption activity to amino groups and adopting an in-situ growth method, after the template molecules are eluted, the original positions of the template molecules are changed into cavities, namely the molecular imprinting polymer of the template molecules is eluted, and therefore, the iron nitride-based macrolide antibiotic sensor is prepared. When the sensor is used for detecting macrolide antibiotic molecules, the macrolide antibiotic sensor based on iron nitride is inserted into a solution to be detected, and the macrolide antibiotic molecules in the solution to be detected are adsorbed into cavities of NIP. The larger the concentration of macrolide antibiotic molecules in the solution to be tested is, the more macrolide antibiotic molecules are adsorbed into the cavities of the NIP. When the electrochemiluminescence detection is carried out, the current intensity passing through the electrode is reduced along with the increase of macrolide antibiotic molecules absorbed into the cavities of the NIP, and the corresponding electrochemiluminescence signal is reduced along with the current intensity, so that the concentration of the macrolide antibiotic molecules in the solution to be detected can be qualitatively and quantitatively determined according to the reduction degree of the electrochemiluminescence light signal intensity.
The technical scheme adopted by the invention is as follows:
1. a preparation method of a macrolide antibiotic sensor based on iron nitride is disclosed, wherein the macrolide antibiotic sensor based on iron nitride is obtained by in-situ growth of a template-free molecularly imprinted polymer NIP on an iron nitride nanosheet array electrode FeN-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 macrolide antibiotic molecule;
2. the preparation method of the iron nitride nanosheet array electrode FeN-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 1-3mmol of nitre hexahydrateIron Fe (NO)3)3·6H2O and 3-9mmol 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 precursor electrode of the ferric hydroxide nanosheet array;
(4) inserting the ferric 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 ferric hydroxide nanosheet array precursor electrode into phosphate buffer solution PBS containing dopamine and ammonium persulfate, 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 ferric nitride nanosheet array electrode FeN-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 phosphate buffer solution PBS containing dopamine and ammonium persulfate: the concentration of dopamine is 2-5mg/mL, the concentration of ammonium persulfate is 3-8mg/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 template-containing molecularly imprinted polymer MIP grown in situ by FeN-nanoarray in the technical scheme 1 comprises the following preparation steps:
(1) respectively weighing 0.25-0.45mmol of template molecules and 3-5mmol of 2-methacrylic acid MAA in an ampoule bottle, adding 8-15mL of acetonitrile, and carrying out ultrasonic treatment for 30min until all the template molecules and the 2-methacrylic acid MAA are dissolved;
(2) adding 15-25mmol 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 FeN-nanoarray prepared in the technical scheme 2 onto a rotary stirrer, and inserting the FeN-nanoarray into the front part of the step (2)In a mixed solution of the precursors in N2Under the temperature of an environment and a water bath of 20-40 ℃, rotationally stirring at the speed of 5-200 r/s, simultaneously dropwise adding 1-3mL of 1mmol/L luminol solution and 1mmol of azobisisobutyronitrile AIBN into the mixed solution at the speed of 1-20 drops/s for initiating polymerization, and obtaining the in-situ grown MIP containing the template molecularly imprinted polymer on the FeN-nanoarray;
4. the preparation steps of the FeN-nanoarray in-situ grown template-free molecularly imprinted polymer NIP 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 FeN-nanoarray and contains the template molecularly imprinted polymer into an eluant, eluting the template molecules for 5-20min 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 iron nitride-based macrolide antibiotic sensor in the technical scheme 1 are as follows: washing the template-free molecularly imprinted polymer NIP which grows on the FeN-nanoarray in situ prepared in the technical scheme 2-4 with deionized water for 2-4 times, and airing at room temperature to prepare the macrolide antibiotic sensor based on the iron nitride;
6. the iron nitride-based macrolide antibiotic sensor prepared by the technical scheme 1-5 is applied to detection of macrolide antibiotic molecules, and comprises the following application steps:
(1) preparing a standard solution: preparing a group of macrolide antibiotic molecular standard solutions with different concentrations including blank standards;
(2) modification of a working electrode: inserting the macrolide antibiotic sensor based on iron nitride as a working electrode into macrolide antibiotic molecular 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: forming a triple electrode by using a saturated calomel electrode as a reference electrode, a platinum wire electrode as a counter electrode and the modified working electrode in the step (2)A polar system connected to an electrochemiluminescence detection device; 15mL of phosphate buffer PBS followed by 1mL of 2mmol/L hydrogen peroxide (H) was added to the cell2O2) A solution; applying cyclic voltage to the assembled working electrode by using a double-order pulse voltammetry to detect the intensity of an optical signal of electrochemiluminescence; the intensity of the response light signal of the blank standard was recorded asA 0The intensity of the response light signal of standard solutions containing macrolide antibiotics at different concentrations is recordedA iThe difference in response to the decrease in optical signal intensity is ΔA = A 0-A i，ΔAMass concentration with macrolide antibiotic standard solutionCWith a linear relationship therebetween, plotting ΔA－CA working curve; the concentration of the phosphate buffer solution PBS is 10mmol/L, and the pH value is 7.4; the parameters of the double-order pulse voltammetry during detection are set as follows: the initial potential is 0V, the pulse potential is 0.9V, the pulse time is 0.1s, and the pulse period is 30 s;
(4) detection of macrolide antibiotics in the sample to be tested: replacing the macrolide antibiotic standard solution in the step (1) with a sample to be tested, and detecting according to the difference delta of the decrease of the intensity of the response light signal in the method in the steps (2) and (3)AAnd working curve to obtain the content of macrolide antibiotics in the sample to be detected;
7. the macrolide antibiotic molecules described in technical schemes 1-6 are one of the following macrolide antibiotic molecules: erythromycin, midecamycin, azithromycin, clarithromycin, roxithromycin, tylosin, and ivermectin.
Advantageous results of the invention
(1) The iron nitride-based macrolide antibiotic sensor is simple to prepare, convenient to operate, low in cost, applicable to portable detection and has market development prospect, and a sample can be quickly, sensitively and selectively detected;
(2) according to the invention, the molecularly imprinted polymer is grown in situ on the FeN-nanoarray electrode 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 FeN-nanoarray electrode, and the FeN-nanoarray electrode has excellent electron transfer capacity, so that the detection sensitivity is greatly improved; on the other hand, the FeN-nanoarray has electrocatalytic activity on hydrogen peroxide, and can realize stable and efficient reaction of a luminol-hydrogen peroxide electrochemiluminescence system without adding horseradish peroxidase, so that the prepared sensor does not need to consider the problem of inactivation of biological enzyme, the use and storage of the sensor can be more stable and the conditions are loose, the signal background is further reduced, the detection sensitivity is improved, and meanwhile, the detection cost is greatly reduced and the environmental pollution is reduced;
(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 iron nitride nanosheet array, a sufficiently thin poly dopamine film is formed and simultaneously the poly dopamine film is uniformly covered on the iron nitride nanosheet array, thereby laying a better foundation for more and better polymerized molecularly imprinted polymers in the next step; then utilizing strong adsorption and connection effects of polydopamine on amino groups rich in the molecularly imprinted polymer, skillfully using FeN-nanoarray as a stirrer, immersing and stirring the mixture 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 FeN-nanoarray by controlling the stirring speed, the dropping speed of a polymerization reaction initiator and the polymerization reaction temperature, so that the FeN-nanoarray can firmly load the molecularly imprinted polymer and luminol on one hand, and the stability and the reproducibility of the prepared electrochemiluminescence 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 film forming thickness and quantitatively coating luminol in situ, and fully improving the sensitivity and detection limit of the molecular imprinting-based electrochemiluminescence sensor.
Detailed Description
EXAMPLE 1 preparation of FeN-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 iron nitrate hexahydrate Fe (NO) was weighed3)3·6H2O 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 precursor electrode of the ferric hydroxide nanosheet array;
(4) inserting the ferric 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 ferric hydroxide nanosheet array precursor electrode into phosphate buffer solution PBS containing dopamine and ammonium persulfate, 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 ferric nitride nanosheet array electrode FeN-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 phosphate buffer solution PBS is 0.1mol/L, and the pH value is 7.2.
EXAMPLE 2 preparation of FeN-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) 2mmol of iron nitrate hexahydrate Fe (NO) was weighed3)3·6H2O 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 precursor electrode of the ferric hydroxide nanosheet array;
(4) inserting the ferric 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 ferric hydroxide nanosheet array precursor electrode into phosphate buffer solution PBS containing dopamine and ammonium persulfate, 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 FeN-nanoarray electrode;
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 phosphate buffer solution PBS is 0.1mol/L, and the pH value is 8.0.
EXAMPLE 3 preparation of FeN-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) 3mmol of iron nitrate hexahydrate Fe (NO) was weighed3)3·6H2O 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 precursor electrode of the ferric hydroxide nanosheet array;
(4) inserting the ferric 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 naturally cooling to room temperature in the ammonia environment, then inserting the ferric hydroxide nanosheet array precursor electrode into phosphate buffer solution PBS containing dopamine and ammonium persulfate, reacting for 6 hours at the temperature of 40 ℃, taking out, and performing immersion washing for 4 times by using deionized water to prepare a ferric nitride nanosheet array electrode FeN-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 phosphate buffer solution PBS is 0.1mol/L, and the pH value is 8.5.
Example 4 preparation of iron nitride based macrolide antibiotic sensor
(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) the FeN-nanoarray prepared in example 1 was inserted into the precursor mixed solution of step (2) while being clamped on a rotary stirrer, and stirred in N2Under the temperature of environment and water bath 20 ℃, stirring in a rotating way at the speed of 200 r/s, simultaneously dripping 1mL of 1mmol/L luminol solution and 1mmol of azobisisobutyronitrile AIBN into the mixed solution at the speed of 1 drop/s to initiate polymerization, and obtaining the in-situ grown MIP containing the template molecularly imprinted polymer on the FeN-nanoarray;
(4) immersing the MIP which is obtained in the step (3) and grows in situ on the FeN-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 air drying at room temperature to obtain the iron nitride-based macrolide antibiotic sensor;
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 of iron nitride based macrolide antibiotic sensor
(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 FeN-nanoarray prepared in the technical scheme 2 onto a rotary stirrer, inserting the FeN-nanoarray into the precursor mixed solution in the step (2), and adding the FeN-nanoarray into the precursor mixed solution in the step (2) to obtain the FeN-nanoarray2Under the temperature of environment and water bath 30 ℃, stirring in a rotating way at the speed of 60 r/s, simultaneously dripping 2 mL of 1mmol/L luminol solution and 1mmol of azobisisobutyronitrile AIBN into the mixed solution at the speed of 10 drops/s to initiate polymerization, and obtaining the in-situ grown MIP containing the template molecularly imprinted polymer on the FeN-nanoarray;
(4) immersing the MIP which is obtained in the step (3) and grows in situ on the FeN-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 air drying at room temperature to obtain the iron nitride-based macrolide antibiotic sensor;
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 of iron nitride based macrolide antibiotic sensor
(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 FeN-nanoarray prepared in the technical scheme 2 onto a rotary stirrer, inserting the FeN-nanoarray into the precursor mixed solution in the step (2), and adding the FeN-nanoarray into the precursor mixed solution in the step (2) to obtain the FeN-nanoarray2Stirring at the temperature of 40 ℃ in an environment and water bath at the speed of 5 revolutions per second, and simultaneously dropwise adding 3mL of 1mmol/L luminol solution and 1mmol of azobisisobutyronitrile AIBN into the mixed solution at the speed of 20 drops per secondInitiating polymerization to obtain in-situ grown MIP containing template molecularly imprinted polymer on the FeN-nanoarray;
(4) immersing the MIP which is obtained in the step (3) and grows in situ on the FeN-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 air-drying at room temperature to obtain the iron nitride-based macrolide antibiotic sensor;
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 iron nitride-based macrolide antibiotic sensors prepared in examples 1 to 6 were applied to the detection of macrolide antibiotic molecules by the following steps:
(1) preparing a standard solution: preparing a group of macrolide antibiotic molecular standard solutions with different concentrations including blank standards;
(2) modification of a working electrode: inserting a macrolide antibiotic sensor based on iron nitride as a working electrode into macrolide antibiotic 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: forming a three-electrode system by using a saturated calomel electrode as a reference electrode, a platinum wire electrode as a counter electrode and the modified working electrode in the step (2), and connecting the three-electrode system to electrochemiluminescence detection equipment; 15mL of phosphate buffer PBS followed by 1mL of 2mmol/L hydrogen peroxide (H) was added to the cell2O2) A solution; applying cyclic voltage to the assembled working electrode by using a double-order pulse voltammetry to detect the intensity of an optical signal of electrochemiluminescence; the intensity of the response light signal of the blank standard was recorded asA 0The intensity of the response light signal of standard solutions containing macrolide antibiotics at different concentrations is recordedA iThe difference in response to the decrease in optical signal intensity is ΔA = A 0-A i，ΔAWith macrolide antibioticsMass concentration of standard solutionCWith a linear relationship therebetween, plotting ΔA－CA working curve; the concentration of the phosphate buffer solution PBS is 10mmol/L, and the pH value is 7.4; the parameters of the double-order pulse voltammetry during detection are set as follows: the initial potential is 0V, the pulse potential is 0.9V, the pulse time is 0.1s, and the pulse period is 30 s;
(4) detection of macrolide antibiotics in the sample to be tested: replacing the macrolide antibiotic standard solution in the step (1) with a sample to be tested, and detecting according to the difference delta of the decrease of the intensity of the response light signal in the method in the steps (2) and (3)AAnd working curve to obtain the content of macrolide antibiotics in the sample to be detected.
Example 8 iron nitride based macrolide antibiotic sensors prepared in examples 1-6, applied to the detection of different macrolide antibiotics according to the detection procedure of example 7, the linear range and detection limit are shown in table 1:
TABLE 1 technical indices for the detection of macrolide antibiotics
Example 9 detection of macrolide antibiotics in pig urine samples
Accurately transferring a pig urine sample, adding a macrolide antibiotic standard solution with a certain mass concentration, taking a pig urine sample without adding macrolide antibiotic as a blank, carrying out a labeling recovery experiment, detecting by using the iron nitride-based macrolide antibiotic sensors prepared in examples 1-6 according to the steps of example 7, and measuring the recovery rate of the macrolide antibiotic in the pig urine sample, wherein the detection results are shown in Table 2:
TABLE 2 detection of macrolide antibiotics in pig urine samples
The detection results in table 2 show that the Relative Standard Deviation (RSD) of the results is less than 3.5%, the average recovery rate is 98.4-101.6%, and the method can be used for detecting multiple macrolide antibiotics in pig urine, and is high in sensitivity, strong in specificity, and accurate and reliable in result.
Example 10 detection of macrolide antibiotics in sheep urine samples
Accurately transferring a certain amount of sheep urine samples, adding a macrolide antibiotic standard solution with a certain mass concentration, taking the sheep urine samples without macrolide antibiotics as blanks, carrying out a labeling recovery experiment, detecting by using the macrolide antibiotic sensors based on iron nitride prepared in examples 1-6 according to the steps of example 7, and determining the recovery rate of the macrolide antibiotics in the sheep urine samples, wherein the detection results are shown in Table 3:
TABLE 3 detection results of macrolide antibiotics in sheep urine samples
The detection results in table 3 show that the Relative Standard Deviation (RSD) of the results is less than 3.2%, the average recovery rate is 98.6-101%, and the method can be used for detecting multiple macrolide antibiotics in sheep urine, and is high in sensitivity, strong in specificity, and accurate and reliable in results.
Claims (5)
1. The preparation method of the macrolide antibiotic sensor based on the iron nitride is characterized in that the macrolide antibiotic sensor based on the iron nitride is obtained by growing a template-free molecularly imprinted polymer NIP in situ on an iron nitride nanosheet array electrode FeN-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 macrolide antibiotic molecule;
the preparation method of the FeN-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 1-3mmol of ferric nitrate hexahydrate Fe (NO)3)3·6H2O and 3-9mmol 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 treated 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 precursor electrode of the ferric hydroxide nanosheet array;
(4) inserting the ferric hydroxide nanosheet array precursor electrode obtained in the step (3) into ammonia water for 5-30 seconds, then 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 ferric hydroxide nanosheet array precursor electrode into phosphate buffer solution PBS containing dopamine and ammonium persulfate, 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 the ferric nitride nanosheet array electrode FeN-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;
the phosphate buffer solution PBS containing dopamine and ammonium persulfate is as follows: the concentration of dopamine is 2-5mg/mL, the concentration of ammonium persulfate is 3-8mg/mL, the concentration of phosphate buffer solution PBS is 0.1mol/L, and the pH value is 7.2-8.5;
the MIP containing the template molecular engram polymer is directly grown on the FeN-nanoarray in situ, and the preparation method comprises the following preparation steps:
(A) respectively weighing 0.25-0.45mmol of template molecules and 3-5mmol of 2-methacrylic acid MAA in an ampoule bottle, adding 8-15mL of acetonitrile, and performing ultrasonic treatment for 30min until all the template molecules and the 2-methacrylic acid MAA are dissolved;
(B) adding 15-25mmol of ethylene glycol dimethacrylate EDMA into the solution obtained in the step (A), and carrying out ultrasonic treatment for 30min until the mixture is uniformly mixed to obtain a precursor mixed solution;
(C) clamping the FeN-nanoarray on a rotary stirrer, inserting the FeN-nanoarray into the precursor mixed solution in the step (B), and adding N2And (2) rotationally stirring at the speed of 5-200 r/s at the temperature of 20-40 ℃ in an environment and a water bath, simultaneously dropwise adding 1-3mL of 1mmol/L luminol solution and 1mmol of azobisisobutyronitrile AIBN into the mixed solution at the speed of 1-20 d/s to initiate polymerization, and obtaining the in-situ grown MIP containing the template molecularly imprinted polymer on the FeN-nanoarray.
2. The method for preparing an iron nitride-based macrolide antibiotic sensor according to claim 1, wherein the template-free molecularly imprinted polymer NIP is prepared by the steps of: immersing the obtained MIP which grows in situ on the FeN-nanoarray and contains the molecular imprinting polymer of the template into an eluant, eluting the molecular of the template for 5-20min at room temperature, and then taking out to obtain the NIP of the molecular imprinting polymer without the template; 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).
3. The method for preparing an iron nitride-based macrolide antibiotic sensor according to claim 2, wherein the iron nitride-based macrolide antibiotic sensor is prepared by the steps of: and (3) washing the prepared template-free molecularly imprinted polymer NIP growing in situ on the FeN-nanoarray for 2-4 times by using deionized water, and airing at room temperature to obtain the iron nitride-based macrolide antibiotic sensor.
4. A method of making an iron nitride based macrolide antibiotic sensor according to any of claims 1-3, said macrolide antibiotic is one of the following macrolide antibiotics: erythromycin, midecamycin, azithromycin, clarithromycin, roxithromycin, tylosin, and ivermectin.
5. Use of an iron nitride based macrolide antibiotic sensor prepared by the process according to any of claims 1 to 3, and the iron nitride based macrolide antibiotic sensor prepared for the detection of macrolide antibiotics, wherein said detection steps are as follows:
(a) preparing a standard solution: preparing a group of macrolide antibiotic standard solutions with different concentrations including blank standards;
(b) modification of a working electrode: taking a macrolide antibiotic sensor based on iron nitride as a working electrode, inserting into macrolide antibiotic standard solutions with different concentrations prepared in the step (a), incubating for 10min, taking out, and washing with deionized water for 3 times;
(c) drawing a working curve: forming a three-electrode system by using a saturated calomel electrode as a reference electrode, a platinum wire electrode as a counter electrode and the modified working electrode in the step (b), and connecting the three-electrode system to electrochemiluminescence detection equipment; adding 15mL of phosphate buffer solution PBS and 1mL of 2mmol/L hydrogen peroxide solution into an electrolytic cell in sequence; applying cyclic voltage to the assembled working electrode by using a double-order pulse voltammetry to detect the intensity of an optical signal of electrochemiluminescence; the intensity of the response light signal of the blank standard is recorded as A0The intensity of the response light signal of the standard solution containing different concentrations of macrolide antibiotics is recorded as AiThe difference in response to the decrease in the optical signal intensity is Δ a ═ a0-AiThe mass concentration C of the macrolides antibiotic standard solution is linearly related to the Delta A, and a Delta A-C working curve is drawn; the concentration of the phosphate buffer solution PBS is 10mmol/L, and the pH value is 7.4; the parameters of the double-order pulse voltammetry during detection are set as follows: the initial potential is 0V, the pulse potential is 0.9V, the pulse time is 0.1s, and the pulse period is 30 s;
(d) detection of macrolide antibiotics in the sample to be tested: replacing the macrolide antibiotic standard solution in the step (a) with a sample to be detected, detecting according to the methods in the steps (b) and (c), and obtaining the content of the macrolide antibiotic in the sample to be detected according to the difference delta A of the reduction of the intensity of the response optical signal and the working curve.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811306512.5A CN109254044B (en) | 2018-11-05 | 2018-11-05 | Preparation method and application of macrolide antibiotic sensor based on FeN |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811306512.5A CN109254044B (en) | 2018-11-05 | 2018-11-05 | Preparation method and application of macrolide antibiotic sensor based on FeN |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109254044A CN109254044A (en) | 2019-01-22 |
| CN109254044B true CN109254044B (en) | 2021-01-08 |
Family
ID=65044329
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811306512.5A Expired - Fee Related CN109254044B (en) | 2018-11-05 | 2018-11-05 | Preparation method and application of macrolide antibiotic sensor based on FeN |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109254044B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109254062B (en) * | 2018-11-05 | 2021-01-29 | 济南大学 | Preparation method and application of macrolide antibiotic molecularly imprinted electrochemical sensor |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105866211A (en) * | 2016-05-30 | 2016-08-17 | 北京师范大学 | Preparation method and application for ampicillin molecular engram sensor |
| CN106324065A (en) * | 2016-08-18 | 2017-01-11 | 济南大学 | Preparation of chemiluminiscence light-induced electrochemical sensor and organic phosphorus pesticide detection |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101507916B (en) * | 2009-02-16 | 2011-01-19 | 西北工业大学 | Preparation method of macrolide antibiotics molecular engram polymer microsphere |
| WO2012009239A2 (en) * | 2010-07-13 | 2012-01-19 | The University Of Houston System | Sensors and separation based on molecular recognition via electropolymerization and colloidal layer templates |
| CN103724655B (en) * | 2013-12-18 | 2015-11-25 | 陕西科技大学 | A kind of method adopting hybrid template to prepare ebormycine molecular imprinted polymer membrane |
| US9893141B2 (en) * | 2015-02-26 | 2018-02-13 | Taiwan Semiconductor Manufacturing Co., Ltd. | Magnetic core, inductor, and method for fabricating the magnetic core |
| CN104829774A (en) * | 2015-04-21 | 2015-08-12 | 华南农业大学 | Azithromycin detection molecular imprinting monolithic micro column and preparation method thereof |
| CN109254062B (en) * | 2018-11-05 | 2021-01-29 | 济南大学 | Preparation method and application of macrolide antibiotic molecularly imprinted electrochemical sensor |
-
2018
- 2018-11-05 CN CN201811306512.5A patent/CN109254044B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105866211A (en) * | 2016-05-30 | 2016-08-17 | 北京师范大学 | Preparation method and application for ampicillin molecular engram sensor |
| CN106324065A (en) * | 2016-08-18 | 2017-01-11 | 济南大学 | Preparation of chemiluminiscence light-induced electrochemical sensor and organic phosphorus pesticide detection |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109254044A (en) | 2019-01-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109342516B (en) | Preparation method and application of sulfonamide molecule electrochemical sensor | |
| CN109254056B (en) | Preparation method and application of tetracyclic antibiotic electrochemiluminescence sensor | |
| CN109254049B (en) | Preparation method and application of ampicillin sensor | |
| CN109254052B (en) | Preparation method and application of electrochemical luminescence sensor for organophosphorus pesticide | |
| CN109254053B (en) | Preparation method and application of environmental estrogen electrochemical analysis sensor | |
| CN109254062B (en) | Preparation method and application of macrolide antibiotic molecularly imprinted electrochemical sensor | |
| CN109307696B (en) | Preparation method and application of molecular imprinting sensing electrode for detecting organochlorine pesticide | |
| CN109254059B (en) | Preparation method and application of tetracyclic antibiotic molecularly imprinted electrochemical sensor | |
| CN109307695B (en) | Preparation method and application of chlordimeform electrochemiluminescence sensor | |
| CN113567521A (en) | Magnetic COF surface molecularly imprinted electrochemical sensor and preparation method and application thereof | |
| CN109254048B (en) | Preparation method and application of nitrofuran antibiotic sensor based on cobalt-nickel oxide | |
| CN109254046B (en) | Nitrofuran antibiotic sensor and preparation method thereof | |
| CN109254044B (en) | Preparation method and application of macrolide antibiotic sensor based on FeN | |
| CN109254058B (en) | Preparation method and application of organophosphorus pesticide sensor based on nickel nitride array | |
| CN109254060B (en) | Clenbuterol electrochemical sensing electrode and preparation method thereof | |
| CN109307698B (en) | Preparation method and application of iron-cobalt nitride sensing electrode for detecting organochlorine pesticide | |
| CN109254054B (en) | Preparation method and application of chlordimeform sensor based on cobalt-based nitride nano array | |
| CN109254050B (en) | Clenbuterol electrochemiluminescence sensor and preparation method thereof | |
| CN109254061B (en) | Preparation method and application of sulfonamide molecule electrochemiluminescence sensor | |
| CN109254057B (en) | Preparation method and application of pyrethroid insecticide electrochemical sensing electrode | |
| CN109254055B (en) | Preparation method and application of ampicillin electrochemiluminescence sensor | |
| CN109307697B (en) | Preparation method and application of electrochemiluminescence sensing electrode for detecting praziquantel | |
| CN109254045B (en) | Cobalt-based nitride sensor for detecting praziquantel and preparation method thereof | |
| CN109254051B (en) | Preparation method and application of environmental estrogen electrochemiluminescence sensor | |
| CN109298034A (en) | Molecular imprinting electrochemical sensor of Selective recognition chloramphenicol and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20210108 Termination date: 20211105 |