CN109254062A - A kind of preparation method and application of macrolide antibiotics molecular engram electrochemical sensor - Google Patents
A kind of preparation method and application of macrolide antibiotics molecular engram electrochemical sensor Download PDFInfo
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Abstract
The invention discloses a kind of preparation methods of macrolide antibiotics molecular engram electrochemical sensor.Belong to Nano-function thin films and chemical biosensor technical field.The present invention is prepared for nitrided iron nano-chip arrays can disposably throw first on electrode, using its big specific surface area and to the high adsorption activity of amino, using the method for growth in situ, directly it is prepared for the poly-dopamine film containing electron mediator in succession and using macrolide antibiotics molecule as the molecularly imprinted polymer of template molecule on nitrided iron nano-chip arrays in succession, after by template molecule elution, the position of template molecule originally has become hole, that is the molecularly imprinted polymer of eluted template molecule, thus, a kind of macrolide antibiotics molecular engram electrochemical sensor just prepares completion.
Description
Technical field
The present invention relates to a kind of preparation method and applications of electrochemical sensor.Belong to Nano-function thin films and chemistry
Biosensor technology field.
Background technique
Macrolide antibiotics (macrolides antibiotics, MA) is that have 12-16 in molecule structure
The general name of the antibacterials of carbon lactone ring inhibits bacterio protein by blocking the activity of peptidy transeferace in 50s ribosomes
Synthesis, belongs to quick bacteriostatic agent.It is mainly used for treating aerobic gram-positive cocci and negative cocci, certain anaerobic bacterias and legion
The infection such as bacterium, mycoplasma, Chlamydia.After people are edible remains the animal food of such antibiotic, animal is generated resistance to
Pharmacological property can also be transferred to the mankind, to endanger human health.Currently, the method for detection macrolide antibiotics molecule mainly has
Enzyme-linked immunization, high performance liquid chromatography, mass spectrography etc..Such method instrument is valuable, complicated for operation, and laboratory personnel needs profession
It just can be carried out detection after training.Therefore, it develops as early as possible a kind of quick, highly selective and Sensitive Detection macrolide antibiotics
Method is extremely important to publilc health, and has wide market application prospect.
Molecular imprinting electrochemical sensor has high specific selectivity, excellent stability, excellent reproducibility, wide inspection
Survey range and floor detection limit.Due to the sensor prepare simple, easy to detect, high sensitivity, it is at low cost the advantages that it is extensive
Applied to the fields such as chromatographic isolation, film point, Solid Phase Extraction, medicine controlled releasing, chemical sensitisation.Molecularly imprinted polymer (MIP), also referred to as
It, being capable of specific recognition and the specific target molecule of selective absorption (i.e. template molecule) for " plastics antibody ".Due to molecular engram
Technology has many advantages that, such as organic reagent corrosion resistance, good stability, heat-resisting quantity and preparation are simple.Therefore, in mistake
In several years gone, electroanalysis is caused based on the MIP electrochemical sensor (MIP-ECS) that MIP is combined with electrochemical sensor
The detection of the focus of chemical field, especially small molecule contaminants.However, having in the preparation process of traditional MIP-ECS
The elution of template molecule difficulty, the disadvantages of thickness of blotting membrane is difficult to control, reproducibility is poor, limit molecular engram film and passed in electrochemistry
Application in sensor.These problems, especially molecular engram film thickness are not easy to control to lead to electrochemical sensor sensitivity decrease
And molecular engram film easily falls off from electrode surface the technical problem for causing stability and reproducibility to reduce during elution, limit
The application of MIP_ECS has been made, therefore, has found new molecularly imprinted polymer synthetic method, new molecular engram film electrode is repaired
The combination method of decorations method and molecular engram film and base material, to solve the preparation of MIP-ECS and have using problem important
Research significance and market value.
Summary of the invention
The purpose of the present invention is to provide a kind of high specificity, prepare simple, easy to detect, high sensitivity, at low cost
The preparation method of macrolide antibiotics molecular engram electrochemical sensor, prepared electrochemical sensor, preparation is simple,
Favorable reproducibility, stability are strong, can be used for quick, the Sensitive Detection of macrolide antibiotics molecule.Based on this purpose, the present invention
It is prepared for nitrided iron nano-chip arrays on electrode can disposably throw first, is inhaled using its big specific surface area and to the height of amino
Attached activity is directly prepared in succession containing electronic media on nitrided iron nano-chip arrays in succession using the method for growth in situ
The poly-dopamine film of body and using macrolide antibiotics molecule as the molecularly imprinted polymer of template molecule, by template point
After son elution, the position of template molecule originally has become hole, the i.e. molecularly imprinted polymer of eluted template molecule, by
This, a kind of macrolide antibiotics molecular engram electrochemical sensor just prepares completion.When for macrolides antibiosis
When plain molecule is detected, macrolide antibiotics molecular engram electrochemical sensor is inserted into solution to be measured, it is to be measured molten
Macrolide antibiotics molecule in liquid can be adsorbed onto the hole of NIP.Macrolide antibiotics point in solution to be measured
Sub- concentration is bigger, and it is more to be adsorbed onto macrolide antibiotics molecule in the hole of NIP.When carrying out Electrochemical Detection, detection
The intensity of electric current can be with increasing and becoming smaller for macrolide antibiotics molecule in the hole of NIP be adsorbed onto, thus according to electricity
The degree that intensity of flow reduces is capable of the concentration of macrolide antibiotics molecule in qualitative, quantitative solution to be measured.
The technical solution adopted by the invention is as follows:
1. a kind of preparation method of macrolide antibiotics molecular engram electrochemical sensor, the macrolides are anti-
Raw element molecular imprinting electrochemical sensor is by growth in situ on nitrided iron nano-chip arrays electrode FeN-nanoarray without template point
What sub- molecularly imprinted polymer NIP was obtained；Described is free from template molecule without template molecule molecularly imprinted polymer NIP
Molecularly imprinted polymer；The molecularly imprinted polymer without containing template molecule is polymerize by molecular engram containing template molecule
What object MIP was obtained by eluted template molecule；The MIP of molecularly imprinted polymer containing template molecule is containing template molecule
Molecularly imprinted polymer；The template molecule is macrolide antibiotics molecule；
2. the preparation method of nitrided iron nano-chip arrays electrode FeN-nanoarray described in technical solution 1 includes following system
Standby step:
(1) it will can disposably throw electrode and carry out ultrasonic cleaning processing using dilute hydrochloric acid, dehydrated alcohol and deionization respectively, to go
Except the oxide layer and surface impurity that can disposably throw electrode；
(2) 1 ~ 3 mmol, six nitric hydrate iron Fe (NO is weighed3)3·6H2O and 3 ~ 9 mmol urea CO (NH2)2, it is put into 50
In mL beaker, 30 mL deionized waters are added and stir to clarify, are then transferred into 50 mL ptfe autoclaves；
(3) in the disposable solution thrown in the reaction kettle that electrode is put into step (2) for handling step (1) well, 100 ~
It is reacted 9 ~ 12 hours at a temperature of 130 DEG C, iron hydroxide nano-chip arrays presoma electrode is prepared；
(4) the iron hydroxide nano-chip arrays presoma electrode that step (3) obtains is inserted into ammonium hydroxide and is taken out after 5 ~ 30 seconds, in ammonia
Under compression ring border, after being heated to 340 ~ 400 DEG C and being kept for 4 ~ 8 hours, continuation naturally rings to room temperature under ammonia environment, then by it
Be inserted into the phosphate buffer solution PBS containing dopamine, Ammonium Persulfate 98.5 and cobalt nitrate in, 20 ~ 40 DEG C at a temperature of reaction 4 ~
After 6 hours, takes out and embathed 2 ~ 4 times with deionized water, nitrided iron nano-chip arrays electrode FeN-nanoarray is prepared；
The described disposable electrode of throwing is selected from one of cycle unit: foamed iron, foam copper, pure iron piece, pure copper sheet, pure iron piece,
Pure silicon piece, conductive carbon cloth；
In the phosphate buffer solution PBS containing dopamine, Ammonium Persulfate 98.5 and cobalt nitrate: dopamine concentration is 2 ~ 5
Mg/mL, the concentration of Ammonium Persulfate 98.5 are 3 ~ 8 mg/mL, and the concentration of cobalt nitrate is 0.1 ~ 0.5 mg/mL, phosphate buffer solution PBS
Concentration be 0.1 mol/L, pH value be 7.2 ~ 8.5；
3. the MIP's of molecularly imprinted polymer containing template molecule of FeN-nanoarray growth in situ described in technical solution 1
Preparation method includes following preparation step:
(1) 0.25 ~ 0.45mmol template molecule and 3 ~ 5 mmol 2- methacrylic acid MAA are weighed respectively in peace times bottle, are added
Enter 8 ~ 15 mL acetonitriles, 30 min of ultrasound to whole dissolutions；
(2) 15 ~ 25 mmol ethylene glycol dimethacrylate EDMA are added in the solution of step (1), 30 min of ultrasound
To being uniformly mixed, precursor mixed solution is obtained；
(3) FeN-nanoarray prepared in technical solution 2 is clipped on Stirring device, the forerunner being inserted into step (2)
In body mixed solution, in N2At a temperature of environment and 20 ~ 40 DEG C of water-bath, with 5 ~ 200 revolutions per seconds of speed Stirring, while with 1
~ 20 drops/sec of speed carries out initiation polymerization to 1 mmol azodiisobutyronitrile AIBN of mixed solution and dripping, in FeN-
The MIP of molecularly imprinted polymer containing template molecule of growth in situ is obtained on nanoarray；
4. FeN-nanoarray growth in situ described in technical solution 1 without template molecule molecularly imprinted polymer NIP's
Preparation step are as follows: by the molecular engram containing template molecule of growth in situ gathers on FeN-nanoarray obtained in technical solution 3
It closes object MIP to be immersed in eluant, eluent, template molecule is subjected to 5 ~ 20 min of elution at room temperature, is then taken out, no mould is obtained
Plate molecular imprinted polymer NIP；The eluant, eluent is the mixed liquor of formic acid and methanol, wherein the volume of formic acid and methanol
Than for 9:(1 ~ 5)；
5. the preparation step of macrolide antibiotics molecular engram electrochemical sensor described in technical solution 1 are as follows: by skill
In art scheme 2 ~ 4 the obtained growth in situ on FeN-nanoarray without template molecule molecularly imprinted polymer NIP, spend
Ionized water embathes 2 ~ 4 times, dries at room temperature, obtains macrolide antibiotics molecular engram electrochemical sensor；
6. being passed using macrolide antibiotics molecular engram electrochemistry prepared by technical solution described in technical solution 1 ~ 5
Sensor, applied to the detection of macrolide antibiotics molecule, including following applying step:
(1) standard solution is prepared: preparing the macrolide antibiotics molecule of one group of various concentration including blank standard specimen
Standard solution；
(2) working electrode is modified: being working electrode, inserting step by macrolide antibiotics molecular engram electrochemical sensor
(1) the macrolide antibiotics molecular criteria solution for the various concentration prepared in takes out after hatching 10 min, uses deionized water
It embathes 3 times；
(3) working curve is drawn: using saturated calomel electrode electrode as reference electrode, platinum electrode is used as to electrode, with step
(2) the working electrode composition three-electrode system modified, connects electrochemical workstation, 15 mL is successively added in a cell
Phosphate buffer solution PBS；Pass through the current-responsive of the working electrode of Differential Pulse Voltammetry DPV detection assembling；Blank standard specimen
Response current intensity be denoted asI 0, the response current intensity of the macrolide antibiotics molecular criteria solution containing various concentration
It is denoted asI i, the difference of response current strength reduction is ΔI = I 0-I i, ΔIWith macrolide antibiotics molecular criteria solution
Mass concentrationCBetween it is linear, draw ΔI?CWorking curve；The phosphate buffer solution PBS concentration is 10
Mmol/L, pH value 7.4；Parameter setting when the described DPV detection are as follows: range and direction are 0 ~ 1 V, and stride is 0.05 V,
Burst length is 0.05 s, and the sampling time is 0.016 s, and the pulse period is 0.5 s；
(4) in sample to be tested macrolide antibiotics molecule detection: replace the macrolide in step (1) with sample to be tested
Class antibiotic molecule standard solution is detected according to the method in step (2) and (3), and current strength reduces according to response
Difference DELTAIAnd working curve, obtain the content of macrolide antibiotics molecule in sample to be tested；
7. macrolide antibiotics molecule described in technical solution 1 ~ 6 is one of following macrolide antibiotics molecule: red
Mycin, medecamycin, azithromycin, clarithromycin, roxithromycin, tylosin, ivermectin.
Beneficial achievement of the invention
(1) macrolide antibiotics molecular engram electrochemical sensor preparation of the present invention is simple, easy to operate, realizes
Quick, sensitive, highly selective detection to sample, and it is at low cost, it can be applied to portable inspectiont, before there is market development
Scape；
(2) the growth in situ molecularly imprinted polymer on nitrided iron nano-chip arrays electrode FeN-nanoarray for the first time of the invention,
On the one hand more, molecularly imprinted polymer more evenly can be grown using the big specific surface area of FeN-nanoarray, and
FeN-nanoarray has excellent electron transmission ability, to greatly improve detection sensitivity；On the other hand, the present invention will
When on dopamine in-situ polymerization to nitrided iron nano-chip arrays, creative is doped into cobalt ions as electron mediator, is examining
Electrochemical response electric current is directly generated when survey, allows sensor straight in the buffer solution it is not necessary that other media substance is added
Capable detection is tapped into, to greatly reduce testing cost simultaneously while further decreasing signal background, improving detection sensitivity
It reduces environmental pollution；
(3) present invention specific surface area and dopamine big using high adsorption activity and nano-array electrode of the nitride to amino
It combines, so that dopamine in nitrided iron nano-chip arrays in situ Polymerization, is forming sufficiently thin poly-dopamine film
While, on uniform fold to nitrided iron nano-chip arrays, thus for more preferably polymerizable molecular imprinted polymers in next step
Carry out place mat；Later using poly-dopamine to the strong absorption connection function for the amino being rich on molecularly imprinted polymer, then ingeniously
It uses FeN-nanoarray as blender wonderfully, immersion stirring is carried out in molecular engram precursor mixed solution, pass through control
The rate of addition and polymeric reaction temperature of mixing speed processed, initiators for polymerization, in the surface FeN-nanoarray direct in-situ
Growth can control the molecularly imprinted polymer of film thickness, print the secured supporting molecular of FeN-nanoarray
Mark polymer, to significantly improve the stability and reproducibility of prepared electrochemical sensor；On the other hand it can effectively control
Molecularly imprinted polymer processed solves in the film forming thickness of electrode surface and the in-stiu coating of electron mediator and is unable to control molecule
Blotting membrane is unable to control the technical problem so as to cause poor reproducibility in electrode surface film forming thickness；In addition, more due to the present invention
Effective control of the preparation method to film forming thickness, can sufficiently improve the sensitive of the electrochemical sensor based on molecular engram
Degree and detection limit have important scientific meaning and application value.
Specific embodiment
The preparation of 1 FeN-nanoarray of embodiment
(1) it will can disposably throw electrode and carry out ultrasonic cleaning processing using dilute hydrochloric acid, dehydrated alcohol and deionization respectively, to go
Except the oxide layer and surface impurity that can disposably throw electrode；
(2) 1 mmol, six nitric hydrate iron Fe (NO is weighed3)3·6H2O and 3 mmol urea CO (NH2)2, it is put into 50 mL
In beaker, 30 mL deionized waters are added and stir to clarify, are then transferred into 50 mL ptfe autoclaves；
(3) in the disposable solution thrown in the reaction kettle that electrode is put into step (2) for handling step (1) well, 100
It is reacted 12 hours at a temperature of DEG C, iron hydroxide nano-chip arrays presoma electrode is prepared；
(4) the iron hydroxide nano-chip arrays presoma electrode that step (3) obtains is inserted into ammonium hydroxide and is taken out after 5 seconds, in ammonia
Under environment, be heated to 340 DEG C and keep 8 hours after, continuation naturally ring to room temperature under ammonia environment, be then inserted into containing
In the phosphate buffer solution PBS of dopamine, Ammonium Persulfate 98.5 and cobalt nitrate, 20 DEG C at a temperature of reaction 4 hours after, take out
And embathed 2 times with deionized water, nitrided iron nano-chip arrays electrode FeN-nanoarray is prepared；
The electrode therein that can disposably throw is foamed iron；Dopamine concentration is 2 mg/mL, and the concentration of Ammonium Persulfate 98.5 is 3 mg/mL,
The concentration of cobalt nitrate is 0.1 mg/mL, and the concentration of phosphate buffer solution PBS is 0.1 mol/L, pH value 7.2.
The preparation of 2 FeN-nanoarray of embodiment
(1) it will can disposably throw electrode and carry out ultrasonic cleaning processing using dilute hydrochloric acid, dehydrated alcohol and deionization respectively, to go
Except the oxide layer and surface impurity that can disposably throw electrode；
(2) 2 mmol, six nitric hydrate iron Fe (NO is weighed3)3·6H2O and 6 mmol urea CO (NH2)2, it is put into 50 mL
In beaker, 30 mL deionized waters are added and stir to clarify, are then transferred into 50 mL ptfe autoclaves；
(3) in the disposable solution thrown in the reaction kettle that electrode is put into step (2) for handling step (1) well, 110
It is reacted 11 hours at a temperature of DEG C, iron hydroxide nano-chip arrays presoma electrode is prepared；
(4) the iron hydroxide nano-chip arrays presoma electrode that step (3) obtains is inserted into ammonium hydroxide and is taken out after 15 seconds, in ammonia
Under environment, be heated to 370 DEG C and keep 6 hours after, continuation naturally ring to room temperature under ammonia environment, be then inserted into containing
In the phosphate buffer solution PBS of dopamine, Ammonium Persulfate 98.5 and cobalt nitrate, 30 DEG C at a temperature of reaction 5 hours after, take out
And embathed 3 times with deionized water, nitrided iron nano-chip arrays electrode FeN-nanoarray is prepared；
The electrode therein that can disposably throw is pure copper sheet；Dopamine concentration is 3.5 mg/mL, and the concentration of Ammonium Persulfate 98.5 is 6.2
Mg/mL, the concentration of cobalt nitrate are 0.3 mg/mL, and the concentration of phosphate buffer solution PBS is 0.1 mol/L, pH value 8.0.
The preparation of 3 FeN-nanoarray of embodiment
(1) it will can disposably throw electrode and carry out ultrasonic cleaning processing using dilute hydrochloric acid, dehydrated alcohol and deionization respectively, to go
Except the oxide layer and surface impurity that can disposably throw electrode；
(2) 3 mmol, six nitric hydrate iron Fe (NO is weighed3)3·6H2O and 9 mmol urea CO (NH2)2, it is put into 50 mL
In beaker, 30 mL deionized waters are added and stir to clarify, are then transferred into 50 mL ptfe autoclaves；
(3) in the disposable solution thrown in the reaction kettle that electrode is put into step (2) for handling step (1) well, 130
It is reacted 9 hours at a temperature of DEG C, iron hydroxide nano-chip arrays presoma electrode is prepared；
(4) the iron hydroxide nano-chip arrays presoma electrode that step (3) obtains is inserted into ammonium hydroxide and is taken out after 30 seconds, in ammonia
Under environment, be heated to 400 DEG C and keep 4 hours after, continuation naturally ring to room temperature under ammonia environment, be then inserted into containing
In the phosphate buffer solution PBS of dopamine, Ammonium Persulfate 98.5 and cobalt nitrate, 40 DEG C at a temperature of reaction 6 hours after, take out
And embathed 4 times with deionized water, nitrided iron nano-chip arrays electrode FeN-nanoarray is prepared；
The electrode therein that can disposably throw is conductive carbon cloth；Dopamine concentration is 5 mg/mL, and the concentration of Ammonium Persulfate 98.5 is 8 mg/
ML, the concentration of cobalt nitrate are 0.5 mg/mL, and the concentration of phosphate buffer solution PBS is 0.1 mol/L, pH value 8.5.
The preparation method of 4 macrolide antibiotics molecular engram electrochemical sensor of embodiment
(1) 0.25 mmol template molecule and 3 mmol 2- methacrylic acid MAA are weighed respectively in peace times bottle, and 8 mL second are added
Nitrile, 30 min of ultrasound to whole dissolutions；
(2) 15 mmol ethylene glycol dimethacrylate EDMA are added in the solution of step (1), 30 min of ultrasound are to mixed
It closes uniformly, obtains precursor mixed solution；
(3) FeN-nanoarray prepared in embodiment 1 is clipped on Stirring device, the presoma being inserted into step (2)
In mixed solution, in N2At a temperature of environment and 20 DEG C of water-bath, with 200 revolutions per seconds of speed Stirring, while with 1 drop/sec
Speed to 1 mmol azodiisobutyronitrile AIBN of mixed solution and dripping carry out initiation polymerization, on FeN-nanoarray
To the MIP of molecularly imprinted polymer containing template molecule of growth in situ；
(4) MIP of molecularly imprinted polymer containing template molecule of growth in situ on FeN-nanoarray for obtaining step (3)
It is immersed in eluant, eluent, template molecule is subjected to 5 min of elution at room temperature, then takes out, obtains no template molecule molecule
Imprinted polymer NIP；Continue to be embathed 2 times with deionized water, dries at room temperature, obtain macrolide antibiotics molecular engram
Electrochemical sensor；
Eluant, eluent therein is the mixed liquor of formic acid and methanol, and wherein the volume ratio of formic acid and methanol is 9:1.
The preparation method of 5 macrolide antibiotics molecular engram electrochemical sensor of embodiment
(1) 0.35mmol template molecule and 4 mmol 2- methacrylic acid MAA are weighed respectively in peace times bottle, and 12 mL second are added
Nitrile, 30 min of ultrasound to whole dissolutions；
(2) 18 mmol ethylene glycol dimethacrylate EDMA are added in the solution of step (1), 30 min of ultrasound are to mixed
It closes uniformly, obtains precursor mixed solution；
(3) FeN-nanoarray prepared in technical solution 2 is clipped on Stirring device, the forerunner being inserted into step (2)
In body mixed solution, in N2At a temperature of environment and 30 DEG C of water-bath, with 60 revolutions per seconds of speed Stirring, while with 10 drops/sec
Speed to 1 mmol azodiisobutyronitrile AIBN of mixed solution and dripping carry out initiation polymerization, on FeN-nanoarray
To the MIP of molecularly imprinted polymer containing template molecule of growth in situ；
(4) MIP of molecularly imprinted polymer containing template molecule of growth in situ on FeN-nanoarray for obtaining step (3)
It is immersed in eluant, eluent, template molecule is subjected to 10 min of elution at room temperature, then takes out, obtains no template molecule molecule
Imprinted polymer NIP；Continue to be embathed 3 times with deionized water, dries at room temperature, obtain macrolide antibiotics molecular engram
Electrochemical sensor；
Eluant, eluent therein is the mixed liquor of formic acid and methanol, and wherein the volume ratio of formic acid and methanol is 9:3.
The preparation method of 6 macrolide antibiotics molecular engram electrochemical sensor of embodiment
(1) 0.45mmol template molecule and 5 mmol 2- methacrylic acid MAA are weighed respectively in peace times bottle, and 15 mL second are added
Nitrile, 30 min of ultrasound to whole dissolutions；
(2) 25 mmol ethylene glycol dimethacrylate EDMA are added in the solution of step (1), 30 min of ultrasound are to mixed
It closes uniformly, obtains precursor mixed solution；
(3) FeN-nanoarray prepared in technical solution 2 is clipped on Stirring device, the forerunner being inserted into step (2)
In body mixed solution, in N2At a temperature of environment and 40 DEG C of water-bath, with 5 revolutions per seconds of speed Stirring, while with 20 drops/sec
Speed to 1 mmol azodiisobutyronitrile AIBN of mixed solution and dripping carry out initiation polymerization, on FeN-nanoarray
To the MIP of molecularly imprinted polymer containing template molecule of growth in situ；
(4) MIP of molecularly imprinted polymer containing template molecule of growth in situ on FeN-nanoarray for obtaining step (3)
It is immersed in eluant, eluent, template molecule is subjected to 20 min of elution at room temperature, then takes out, obtains no template molecule molecule
Imprinted polymer NIP；Continue to be embathed 4 times with deionized water, dries at room temperature, obtain macrolide antibiotics molecular engram
Electrochemical sensor；
Eluant, eluent therein is the mixed liquor of formic acid and methanol, and wherein the volume ratio of formic acid and methanol is 9:5.
The macrolide antibiotics molecular engram electrochemical sensor of 7 embodiment 1 ~ 6 of embodiment preparation is applied to big
The detection of cyclic lactone class antibiotic molecule, steps are as follows:
(1) standard solution is prepared: preparing the macrolide antibiotics molecule of one group of various concentration including blank standard specimen
Standard solution；
(2) working electrode is modified: being working electrode, inserting step by macrolide antibiotics molecular engram electrochemical sensor
(1) the macrolide antibiotics standard solution for the various concentration prepared in takes out after hatching 10 min, is embathed with deionized water
3 times；
(3) working curve is drawn: using saturated calomel electrode electrode as reference electrode, platinum electrode is used as to electrode, with step
(2) the working electrode composition three-electrode system modified, connects electrochemical workstation, 15 mL is successively added in a cell
PBS；Pass through the current-responsive of the working electrode of Differential Pulse Voltammetry DPV detection assembling；The response current intensity of blank standard specimen
It is denoted asI 0, the response current intensity of the macrolide antibiotics molecular criteria solution containing various concentration is denoted asI i, response current
The difference of strength reduction is ΔI = I 0-I i, ΔIWith the mass concentration of macrolide antibiotics standard solutionCBetween it is linear
Relationship draws ΔI?CWorking curve；The PBS is the phosphate buffer solution of 10 mmol/L, the phosphate-buffered
The pH value of solution is 7.4；Parameter setting when the described DPV detection are as follows: range and direction are 0 ~ 1 V, and stride is 0.05 V, arteries and veins
Rushing the time is 0.05 s, and the sampling time is 0.016 s, and the pulse period is 0.5 s；
(4) in sample to be tested macrolide antibiotics detection: replace the macrolides in step (1) anti-with sample to be tested
Raw element standard solution, is detected, the difference DELTA that current strength reduces according to response according to the method in step (2) and (3)IWith
Working curve obtains the content of macrolide antibiotics in sample to be tested.
The macrolide antibiotics molecular engram electrochemical sensor of 8 embodiment 1 ~ 6 of embodiment preparation, according to implementation
The detecting step of example 7 is applied to the detection of different macrolide antibiotics, and the range of linearity and detection limit are shown in Table 1:
The detection technique index of 1 macrolide antibiotics of table
The detection of macrolide antibiotics in 9 pig urine samples of embodiment
Pig urine samples are accurately pipetted, the macrolide antibiotics standard solution of certain mass concentration are added, big ring not to be added
The pig urine samples of lactone antibiotic are blank, carry out recovery testu, the macrolides antibiosis prepared with embodiment 1 ~ 6
Plain molecular imprinting electrochemical sensor is detected according to the step of embodiment 7, measures macrolides antibiosis in pig urine samples
The rate of recovery of element, testing result are shown in Table 2:
The testing result of macrolide antibiotics in 2 pig urine samples of table
For 2 testing result of table it is found that the relative standard deviation (RSD) of result is less than 3.2 %, average recovery rate is 98.2 ~ 101%,
Show the present invention can be used for pig urine in a variety of macrolide antibiotics detection, the high sensitivity of method, high specificity, as a result
Accurately and reliably.
The detection of macrolide antibiotics in 10 sheep urine samples of embodiment
Certain sheep urine samples are accurately pipetted, the macrolide antibiotics standard solution of certain mass concentration are added, not to be added
The sheep urine samples of macrolide antibiotics are blank, carry out recovery testu, the macrolides prepared with embodiment 1 ~ 6
Antibiotics molecular engram electrochemical sensor is detected according to the step of embodiment 7, measures macrolides in sheep urine samples
The rate of recovery of antibiotic, testing result are shown in Table 3:
The testing result of macrolide antibiotics in 3 sheep urine samples of table
3 testing result of table it is found that the relative standard deviation (RSD) of result less than 3.5 %, average recovery rate is 98.0 ~
101.4%, show that the present invention can be used for the detection of a variety of macrolide antibiotics in sheep urine, high sensitivity, the specificity of method
By force, as a result accurately and reliably.
Claims (7)
1. a kind of preparation method of macrolide antibiotics molecular engram electrochemical sensor, which is characterized in that described is big
Cyclic lactone class antibiotics molecular engram electrochemical sensor is by situ raw on nitrided iron nano-chip arrays electrode FeN-nanoarray
What length was obtained without template molecule molecularly imprinted polymer NIP；Described has been free from without template molecule molecularly imprinted polymer NIP
The molecularly imprinted polymer of template molecule；The molecularly imprinted polymer without containing template molecule is by containing template molecule point
What sub- imprinted polymer MIP was obtained by eluted template molecule；The MIP of molecularly imprinted polymer containing template molecule be containing
The molecularly imprinted polymer of template molecule；The template molecule is macrolide antibiotics molecule.
2. nitrided iron nano-chip arrays electrode FeN-nanoarray as described in claim 1 is it is characterized in that, the FeN-
The preparation method of nanoarray includes following preparation step:
(1) it will can disposably throw electrode and carry out ultrasonic cleaning processing using dilute hydrochloric acid, dehydrated alcohol and deionization respectively, to go
Except the oxide layer and surface impurity that can disposably throw electrode；
(2) 1 ~ 3 mmol, six nitric hydrate iron Fe (NO is weighed3)3·6H2O and 3 ~ 9 mmol urea CO (NH2)2, it is put into 50
In mL beaker, 30 mL deionized waters are added and stir to clarify, are then transferred into 50 mL ptfe autoclaves；
(3) in the disposable solution thrown in the reaction kettle that electrode is put into step (2) for handling step (1) well, 100 ~
It is reacted 9 ~ 12 hours at a temperature of 130 DEG C, iron hydroxide nano-chip arrays presoma electrode is prepared；
(4) the iron hydroxide nano-chip arrays presoma electrode that step (3) obtains is inserted into ammonium hydroxide and is taken out after 5 ~ 30 seconds, in ammonia
Under compression ring border, after being heated to 340 ~ 400 DEG C and being kept for 4 ~ 8 hours, continuation naturally rings to room temperature under ammonia environment, then by it
Be inserted into the phosphate buffer solution PBS containing dopamine, Ammonium Persulfate 98.5 and cobalt nitrate in, 20 ~ 40 DEG C at a temperature of reaction 4 ~
After 6 hours, takes out and embathed 2 ~ 4 times with deionized water, nitrided iron nano-chip arrays electrode FeN-nanoarray is prepared；
The described disposable electrode of throwing is selected from one of cycle unit: nickel foam, foam copper, pure nickel piece, pure copper sheet, pure iron piece,
Pure silicon piece, conductive carbon cloth；
In the phosphate buffer solution PBS containing dopamine, Ammonium Persulfate 98.5 and cobalt nitrate: dopamine concentration is 2 ~ 5
Mg/mL, the concentration of Ammonium Persulfate 98.5 are 3 ~ 8 mg/mL, and the concentration of cobalt nitrate is 0.1 ~ 0.5 mg/mL, phosphate buffer solution PBS
Concentration be 0.1 mol/L, pH value be 7.2 ~ 8.5.
3. the MIP of molecularly imprinted polymer containing template molecule as described in claim 1, which is characterized in that described containing template point
Sub- molecularly imprinted polymer MIP is that direct in-situ is grown on FeN-nanoarray, and preparation method includes following preparation step
It is rapid:
(1) 0.25 ~ 0.45mmol template molecule and 3 ~ 5 mmol 2- methacrylic acid MAA are weighed respectively in peace times bottle, are added
Enter 8 ~ 15 mL acetonitriles, 30 min of ultrasound to whole dissolutions；
(2) 15 ~ 25 mmol ethylene glycol dimethacrylate EDMA are added in the solution of step (1), 30 min of ultrasound
To being uniformly mixed, precursor mixed solution is obtained；
(3) FeN-nanoarray is clipped on Stirring device, is inserted into the precursor mixed solution in step (2), in N2
At a temperature of environment and 20 ~ 40 DEG C of water-bath, with 5 ~ 200 revolutions per seconds of speed Stirring, while with 1 ~ 20 drop/sec of speed to
1 mmol azodiisobutyronitrile AIBN of mixed solution and dripping carries out initiation polymerization, and life in situ is obtained on FeN-nanoarray
The long MIP of molecularly imprinted polymer containing template molecule.
4. as described in claim 1 without template molecule molecularly imprinted polymer NIP, it is characterised in that the no template point
The preparation step of sub- molecularly imprinted polymer NIP are as follows: by obtained in claim 3 on FeN-nanoarray growth in situ
The MIP of molecularly imprinted polymer containing template molecule be immersed in eluant, eluent, template molecule is subjected to elution 5 ~ 20 at room temperature
Min then takes out, and obtains no template molecule molecularly imprinted polymer NIP；The eluant, eluent is the mixing of formic acid and methanol
Liquid, wherein the volume ratio of formic acid and methanol is 9:(1 ~ 5).
5. the preparation step of macrolide antibiotics molecular engram electrochemical sensor as described in claim 1 are as follows: will weigh
Benefit require in 2 ~ 4 the growth in situ obtained on FeN-nanoarray without template molecule molecularly imprinted polymer NIP, spend
Ionized water embathes 2 ~ 4 times, dries at room temperature, obtains macrolide antibiotics molecular engram electrochemical sensor.
6. using macrolide antibiotics molecular engram electrochemical sensing prepared by preparation method described in claim 1 ~ 5
Device, the detection applied to macrolide antibiotics, which is characterized in that the detecting step is as follows:
(1) standard solution is prepared: preparing the macrolide antibiotics standard of one group of various concentration including blank standard specimen
Solution；
(2) working electrode is modified: being working electrode, inserting step by macrolide antibiotics molecular engram electrochemical sensor
(1) the macrolide antibiotics standard solution for the various concentration prepared in takes out after hatching 10 min, is embathed with deionized water
3 times；
(3) working curve is drawn: using saturated calomel electrode electrode as reference electrode, platinum electrode is used as to electrode, with step
(2) the working electrode composition three-electrode system modified, connects electrochemical workstation, 15 mL is successively added in a cell
Phosphate buffer solution PBS；Pass through the current-responsive of the working electrode of Differential Pulse Voltammetry DPV detection assembling；Blank standard specimen
Response current intensity be denoted asI 0, the response current intensity of the macrolide antibiotics standard solution containing various concentration is denoted asI i, the difference of response current strength reduction is ΔI = I 0-I i, ΔIWith the mass concentration of macrolide antibiotics standard solutionCBetween it is linear, draw ΔI?CWorking curve；The phosphate buffer solution PBS concentration is 10 mmol/L,
PH value is 7.4；Parameter setting when the described DPV detection are as follows: range and direction are 0 ~ 1 V, and stride is 0.05 V, burst length
For 0.05 s, the sampling time is 0.016 s, and the pulse period is 0.5 s；
(4) in sample to be tested macrolide antibiotics detection: replace the macrolides in step (1) anti-with sample to be tested
Raw element standard solution, is detected, the difference DELTA that current strength reduces according to response according to the method in step (2) and (3)IWith
Working curve obtains the content of macrolide antibiotics in sample to be tested.
7. the macrolide antibiotics as described in claim 1 ~ 6 is one of following macrolide antibiotics: erythromycin, wheat
Enlightening mycin, azithromycin, clarithromycin, roxithromycin, tylosin, ivermectin.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109254044A (en) * | 2018-11-05 | 2019-01-22 | 济南大学 | A kind of preparation method and application of the macrolide antibiotics sensor based on nitrided iron |
| CN110204735A (en) * | 2019-05-31 | 2019-09-06 | 中国药科大学 | A kind of preparation method and application of the hollow porous type molecularly imprinted polymer satellite assembly of the magnetic core-of macrolide antibiotics |
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| CN101507916A (en) * | 2009-02-16 | 2009-08-19 | 西北工业大学 | Preparation method of macrolide antibiotics molecular engram polymer microsphere |
| CN101961662A (en) * | 2010-07-29 | 2011-02-02 | 江苏大学 | Method for preparing ion imprinting supported composite photocatalyst |
| WO2012151563A3 (en) * | 2011-05-04 | 2013-03-21 | Dxupclose | Device and method for identifying microbes and counting microbes and determining antimicrobial sensitivity |
| SE1330067A1 (en) * | 2013-06-09 | 2014-12-10 | Carriers for the binding and release of molecules and their method of preparation | |
| CN109254044A (en) * | 2018-11-05 | 2019-01-22 | 济南大学 | A kind of preparation method and application of the macrolide antibiotics sensor based on nitrided iron |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101507916A (en) * | 2009-02-16 | 2009-08-19 | 西北工业大学 | Preparation method of macrolide antibiotics molecular engram polymer microsphere |
| CN101961662A (en) * | 2010-07-29 | 2011-02-02 | 江苏大学 | Method for preparing ion imprinting supported composite photocatalyst |
| WO2012151563A3 (en) * | 2011-05-04 | 2013-03-21 | Dxupclose | Device and method for identifying microbes and counting microbes and determining antimicrobial sensitivity |
| SE1330067A1 (en) * | 2013-06-09 | 2014-12-10 | Carriers for the binding and release of molecules and their method of preparation | |
| CN109254044A (en) * | 2018-11-05 | 2019-01-22 | 济南大学 | A kind of preparation method and application of the macrolide antibiotics sensor based on nitrided iron |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109254044A (en) * | 2018-11-05 | 2019-01-22 | 济南大学 | A kind of preparation method and application of the macrolide antibiotics sensor based on nitrided iron |
| CN110204735A (en) * | 2019-05-31 | 2019-09-06 | 中国药科大学 | A kind of preparation method and application of the hollow porous type molecularly imprinted polymer satellite assembly of the magnetic core-of macrolide antibiotics |
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