CN104020202A - Method for detecting content of clenbuterol in solution to be detected - Google Patents
Method for detecting content of clenbuterol in solution to be detected Download PDFInfo
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- CN104020202A CN104020202A CN201410300939.XA CN201410300939A CN104020202A CN 104020202 A CN104020202 A CN 104020202A CN 201410300939 A CN201410300939 A CN 201410300939A CN 104020202 A CN104020202 A CN 104020202A
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Abstract
The invention discloses a method for detecting the content of clenbuterol in a solution to be detected. The method comprises the following steps; modifying a working electrode by using melamine molecules, synthesizing nano grains wrapped by melamine molecules, fixing the nano grains on the surface of the electrode through the hydrogen bond function of clenbuterol molecules and a plurality of melamine molecules, and obtaining specific electrochemical signals according to electrochemical properties, catalytic properties or other properties of the nano grains, thereby confirming the concentration of the melamine molecules. The method is simple and convenient, high in detection flexibility, low in detection cost and is expected to realize large-scale popularization and application.
Description
Technical field
The present invention relates to electrochemical measuring technique field, particularly a kind of electrochemical detection method that detects clenbuterol hydrochloride content in solution to be measured.
Background technology
Clenbuterol hydrochloride is a class β
2-receptor stimulating agent, can selective excitement β
2-adrenocepter, as Ractopamine, the special human relations in Crow, salbutamol etc.This quasi-molecule bronchial smooth muscle that can relax, mast cell discharges the inflammatory mediators such as histamine always.Therefore, clinically, can use it for the treatment of the diseases such as bronchial astehma.But they also have certain toxic and side effect, may cause myocardial ischemia or arrhythmia cordis, skeletal muscle chatter, the symptoms such as jump back bronchial spasm.
Due to β
2-receptor stimulating agent can promote growth of animal, cause the synthetic increase of protein, significantly improve lean meat percentage, therefore extended to various countries, as animal growth promoting agent, use, be commonly called as " clenbuterol hydrochloride ", but because it easily accumulates residual in animal body, easily cause food security accident, therefore in China, forbidden the illegal β such as the special human relations in Crow that abuse in animal farming industry
2-receptor stimulating agent.2002, China Supreme People's Court and Supreme People's Procuratorate's issue bulletin (method is released [2002] No. 26), clear and definite " clenbuterol hydrochloride " illegal production, the criterion for imposing penalty of selling and using.Then, still there are at present many illegal operators using clenbuterol hydrochloride.
For the detection method of clenbuterol hydrochloride, mainly divide at present instrument confirmation method and the large class of immune analysis method two.Instrument confirmation method comprises LC-MS-MS, gas chromatography-mass spectrography technology, high performance liquid chromatography, capillary electrophoresis etc.Immune analysis method is mainly the specific binding based on Ag-Ab, comprises immunochromatographic method, electrochemical immunosensor etc.These methods have weak point, and they depend on large-scale instrument, depend on expensive reagent, therefore still need exploitation to facilitate the sensitive new method that detects cheaply clenbuterol hydrochloride.
Summary of the invention
For above-mentioned weak point, the object of the present invention is to provide a kind of electrochemical detection method based on melamine molecule and the identification of clenbuterol hydrochloride molecular specificity easy and simple to handle, highly sensitive, with low cost.
The invention provides a kind of method for detection of clenbuterol hydrochloride content in solution to be measured, comprise the following steps:
Step 1) melamine molecule is modified in a plurality of working electrode surfaces respectively;
Step 2) synthesis of nano particle, and melamine molecule is modified in nano grain surface, subsequently nano particle is made into the solution for later use that concentration is 1～50nM;
Step 3) working electrode that respectively each is modified with to melamine molecule immerses in the clenbuterol hydrochloride titer of three or three above concentration known, a plurality of hydrogen bonds of the intermolecular formation of melamine molecule and clenbuterol hydrochloride, thus by clenbuterol hydrochloride molecule trapping to working electrode surface;
Step 4) by step 3) each of gained working electrode of catching clenbuterol hydrochloride molecule immerses respectively step 2) in the nanoparticles solution of gained, the vacant group of the melamine molecule of nano grain surface and the clenbuterol hydrochloride molecular moiety on working electrode forms hydrogen bond, thereby nano particle is fixed on to working electrode surface;
Step 5) detect the electrochemical response that each is fixed with the working electrode of nano material, determine the concentration of clenbuterol hydrochloride titer and the relation of electrochemical signals, draw the canonical plotting of clenbuterol hydrochloride concentration and electrochemical signals;
Step 6) by step 1) working electrode that is modified with melamine molecule that obtains immerses solution to be measured and step 2 successively) in the nanoparticles solution of gained, the electrochemical response of testing electrode, thus and know the content of clenbuterol hydrochloride in solution to be measured with canonical plotting comparison.
Preferably, the described method for detection of clenbuterol hydrochloride content in solution to be measured, described working electrode is gold electrode or carbon electrode.
Preferably, the described method for detection of clenbuterol hydrochloride content in solution to be measured, described step 2) in nano particle be semiconductor-quantum-point or metal nanoparticle.
Preferably, the described method for detection of clenbuterol hydrochloride content in solution to be measured, described metal nanoparticle is gold nano grain or silver nano-grain or Pt nanoparticle.
Preferably, the described method for detection of clenbuterol hydrochloride content in solution to be measured, described step 2) in the synthetic method of nano particle be hydro-thermal method or microwave method.
Preferably, the described method for detection of clenbuterol hydrochloride content in solution to be measured, described step 2) in nano particle after synthesizing, before being modified with melamine molecule, carry out purification process, purification process is centrifugal or dialysis.
Preferably, the described method for detection of clenbuterol hydrochloride content in solution to be measured, described electrochemical response records by cyclic voltammetry or differential pulse voltammetry or AC impedence method or linear sweep voltammetry.
The invention has the beneficial effects as follows: melamine-clenbuterol hydrochloride molecular recognition system and electrochemical techniques are organically combined, quantitative detection for clenbuterol hydrochloride molecule, not only detection sensitivity is high, easy to operate for it, also overcome the dependence to large-scale expensive instrument, avoided the use of the expensive reagent such as antigen-antibody, greatly reduce testing cost, contributed to the early diagnosis of relevant disease.
Accompanying drawing explanation
Fig. 1 is the process flow diagram for detection of the electrochemical detection method of clenbuterol hydrochloride content in solution to be measured of the present invention, and wherein, in figure, rectangular of below represents electrode.
Fig. 2 is that the present invention adopts cyclic voltammetry gained testing result figure, wherein, (a) melamine molecule modified electrode directly soaks nanoparticles solution, (b) melamine molecule modified electrode and first soaks and soak the cyclic voltammogram that nanoparticles solution, (c) only have melamine molecule modified electrode after the special human relations solution in 1nM Crow.
Fig. 3 is that the present invention adopts linear sweep voltammetry to detect the linear sweep voltammetry figure of the special human relations in Crow of variable concentrations.
Fig. 4 is the concentration curve of the special human relations of the measured electrochemical signals of the present invention and Crow.
Fig. 5 is the electrochemical response value of the different material surveyed of the present invention, and it has illustrated the antijamming capability of this electrochemical detection method.
Embodiment
Below in conjunction with accompanying drawing, the present invention is described in further detail, to make those skilled in the art can implement according to this with reference to instructions word.
Refer to Fig. 1, it has represented main-process stream of the present invention, the invention provides a kind of method for detection of clenbuterol hydrochloride content in solution to be measured, comprises the following steps:
Step 1) respectively melamine molecule is modified in a plurality of working electrode surfaces stand-byly, working electrode is preferably gold electrode or carbon electrode; Melamine molecule can with golden self assembly, can there is covalent reaction with the metal surface such as gold, silver in the amino of melamine; When using carbon electrode as working electrode, first to carbon electrodes, modify, carry out activation processing, as can be first soaked in concentrated acid, surface is carboxylated, subsequently with melamine in amino condensation; Or adopt the method for carboxylated Graphene physisorption to carry out modified graphite electrode.
Step 2) synthesis of nano particle, and melamine molecule is modified in nano grain surface, subsequently that nano particle wiring solution-forming is stand-by; Nano particle is preferably semiconductor-quantum-point or metal nanoparticle, and metal nanoparticle is preferably gold nano grain or silver nano-grain or Pt nanoparticle.The synthetic method of nano particle is preferably hydro-thermal method or microwave method.Nano particle preferably carries out purification process after synthesizing, before being modified with melamine molecule, and purification process is centrifugal or dialysis, to guarantee the purity of nano particle, improves accuracy of detection.It is same as above that melamine is modified the principle of nano grain surface, melamine can with nano particle self assembly, the amino of melamine can with semiconductor-quantum-point or metal nanoparticle on metal generation covalent reaction.
Step 3) respectively by step 1) each working electrode that is modified with melamine molecule of obtaining immerses in the clenbuterol hydrochloride titer of three or three above concentration known, melamine molecule can with a plurality of hydrogen bonds of the intermolecular formation of clenbuterol hydrochloride, thereby by clenbuterol hydrochloride molecule trapping to working electrode surface.
Step 4) by step 3) each of gained working electrode of catching clenbuterol hydrochloride molecule immerses respectively step 2) in the nanoparticles solution of gained, the vacant group of the melamine molecule of nano grain surface and the clenbuterol hydrochloride molecular moiety on working electrode forms hydrogen bond, thereby nano particle is fixed on to working electrode surface.
Step 5) detect the electrochemical response that each is fixed with the working electrode of nano material, determine the concentration of clenbuterol hydrochloride titer and the relation of electrochemical signals, draw the canonical plotting of clenbuterol hydrochloride concentration and electrochemical signals; When nano particle is fixed on after working electrode surface, the electrochemical environment of electrode surface changes, thereby the electrochemical response performance of electrode is changed, and the content of this change and nano particle, further say with the content of clenbuterol hydrochloride and be proportionate, this case utilizes this point to propose the new method that clenbuterol hydrochloride detects just.Electrochemical response can preferably record by cyclic voltammetry, differential pulse voltammetry, AC impedence method or linear sweep voltammetry.Electrochemical signals its essence is the process of reaction substrate receiving and losing electrons, and nano particle can serve as the effect of catalyzer in the process of reaction substrate receiving and losing electrons, for example, when using Pt nanoparticle, Pt nanoparticle is fixed on electrode surface, working electrode inserts in hydrogen peroxide solution, Pt nanoparticle just can be brought into play the function of catalyzer, is water and oxygen, in this process decomposing hydrogen dioxide solution, the process of electron exchange can be captured by working electrode, thereby produces electrochemical response.Same, when using silver nano-grain, electrochemical signals comes from the solid-state Ag/AgCl course of reaction of silver nano-grain.
Step 6) by step 1) working electrode that is modified with melamine molecule that obtains immerses solution to be measured and step 2 successively) in the nanoparticles solution of gained, the electrochemical response of testing electrode, thus and know the content of clenbuterol hydrochloride in solution to be measured with canonical plotting comparison.
Embodiment
The electrode using in the present embodiment is gold electrode, and nano particle is silver nano-grain, and clenbuterol hydrochloride is the special human relations in Crow.
1.1, melamine molecule is modified the preparation of working electrode
A, polishing gold electrode, step comprises that sand paper grinds, and the alundum (Al2O3) slurry of variable grain size grinds, and ethanol/water is ultrasonic, 50% nitric acid dousing, 0.5M sulfuric acid electrochemical cleaning etc.
B, gold electrode are soaked in 1mM melamine solution, react 6 hours, then washed with de-ionized water electrode.
1.2, silver nano-grain preparation
Silver nitrate, citric acid three sodium solution that preparation 100mL concentration is 0.25mM, and be that 10mM sodium borohydride solution mixes with 3mL concentration, vigorous stirring 30 minutes, then standing over night, centrifugal 30 minutes of 12000g, remove impurity, obtain the silver nano-grain that surface coverage has the trisodium citrate of peptizaiton.Then mix with melamine, its concentration is 0.1mM, reacts 24 hours, and centrifugal 30 minutes of last 12000g, removes impurity.
1.3, typical curve processed
Compound concentration is respectively 0,0.01,0.0464,0.215,1.0,4.64,21.5, the clenbuterol hydrochloride of 100nM (the special human relations in Crow) titer, melamine molecule modified electrode is soaked in clenbuterol hydrochloride titer, hatch 30 minutes, electrode clean is clean, subsequently electrode is soaked in the silver nano-grain after modification, reacts 30 minutes and clean up.Above-mentioned electrode is as working electrode, uses platinum electrode as to electrode, and saturated calomel electrode, as contrast electrode, builds three-electrode system.Utilize linear sweep voltammetry, record signal, multi-group data is made to electric signal-concentration standard curve figure.
1.4, detect the special human relations in Crow
Melamine molecule modified electrode is soaked in solution to be detected, hatches 30 minutes, clean up, be soaked into subsequently in the silver nano-grain after modification, react 30 minutes and clean up.Above-mentioned electrode is as working electrode, uses platinum electrode as to electrode, and saturated calomel electrode, as contrast electrode, builds three-electrode system.Utilize linear sweep voltammetry, record signal, after contrasting with canonical plotting, draw the concentration of the special human relations in Crow.
The explanation of collection of illustrative plates characterization result:
Fig. 2 is that the present invention adopts cyclic voltammetry gained testing result figure, wherein, (a) melamine molecule modified electrode directly soaks nanoparticles solution, (b) melamine molecule modified electrode and first soaks and soak the cyclic voltammogram that nanoparticles solution, (c) only have melamine molecule modified electrode after the special human relations solution in 1nM Crow.As seen from the figure, only in Crow, special human relations exist in situation, after silver nano-grain is fixed on to electrode surface, just have obvious current peak (Ag/AgCl course of reaction) in linear sweep voltammetry figure.
Fig. 3 is that the present invention adopts linear sweep voltammetry to detect the linear sweep voltammetry figure of the special human relations in Crow of variable concentrations, in figure curve represent successively from top to bottom 0,0.01,0.0464,0.215,1.0,4.64,21.5, the special human relations in Crow of 100nM.As seen from the figure, the special human relations concentration of clo is larger, and the silver nano-grain that is fixed on electrode surface is more, thereby the peak point current obtaining is also larger.
Fig. 4 is the concentration curve of the special human relations of the measured electrochemical signals of the present invention and Crow.As seen from the figure, in certain concentration range, the logarithm value of the special human relations concentration in peak point current and Crow is linear dependence.
Fig. 5 is the electrochemical response value of the different material surveyed of the present invention, and it has illustrated the antijamming capability of this electrochemical detection method.As seen from the figure, the material such as carbohydrate, salt, protein, amino acid does not cause obvious interference for the detection of clenbuterol hydrochloride molecule.
Although embodiment of the present invention are open as above, but it is not restricted to listed utilization in instructions and embodiment, it can be applied to various applicable the field of the invention completely, for those skilled in the art, can easily realize other modification, therefore do not deviating under the universal that claim and equivalency range limit, the present invention is not limited to specific details and illustrates here and the legend of describing.
Claims (7)
1. for detection of a method for clenbuterol hydrochloride content in solution to be measured, it is characterized in that, comprise the following steps:
Step 1) melamine molecule is modified in a plurality of working electrode surfaces respectively;
Step 2) synthesis of nano particle, and melamine molecule is modified in nano grain surface, subsequently nano particle is made into the solution for later use that concentration is 1～50nM;
Step 3) working electrode that respectively each is modified with to melamine molecule immerses in the clenbuterol hydrochloride titer of three or three above concentration known, a plurality of hydrogen bonds of the intermolecular formation of melamine molecule and clenbuterol hydrochloride, thus by clenbuterol hydrochloride molecule trapping to working electrode surface;
Step 4) by step 3) each of gained working electrode of catching clenbuterol hydrochloride molecule immerses respectively step 2) in the nanoparticles solution of gained, the vacant group of the melamine molecule of nano grain surface and the clenbuterol hydrochloride molecular moiety on working electrode forms hydrogen bond, thereby nano particle is fixed on to working electrode surface;
Step 5) detect the electrochemical response that each is fixed with the working electrode of nano material, determine the concentration of clenbuterol hydrochloride titer and the relation of electrochemical signals, draw the canonical plotting of clenbuterol hydrochloride concentration and electrochemical signals;
Step 6) by step 1) working electrode that is modified with melamine molecule that obtains immerses solution to be measured and step 2 successively) in the nanoparticles solution of gained, the electrochemical response of testing electrode, thus and know the content of clenbuterol hydrochloride in solution to be measured with canonical plotting comparison.
2. the method for detection of clenbuterol hydrochloride content in solution to be measured according to claim 1, is characterized in that, described working electrode is gold electrode or carbon electrode.
3. the method for detection of clenbuterol hydrochloride content in solution to be measured according to claim 1, is characterized in that described step 2) in nano particle be semiconductor-quantum-point or metal nanoparticle.
4. the method for detection of clenbuterol hydrochloride content in solution to be measured according to claim 3, is characterized in that, described metal nanoparticle is gold nano grain or silver nano-grain or Pt nanoparticle.
5. the method for detection of clenbuterol hydrochloride content in solution to be measured according to claim 1, is characterized in that described step 2) in the synthetic method of nano particle be hydro-thermal method or microwave method.
6. the method for detection of clenbuterol hydrochloride content in solution to be measured according to claim 1, it is characterized in that, described step 2) nano particle in carries out purification process after synthesizing, before being modified with melamine molecule, and purification process is centrifugal or dialysis.
7. the method for detection of clenbuterol hydrochloride content in solution to be measured according to claim 1, is characterized in that, described electrochemical response records by cyclic voltammetry or differential pulse voltammetry or AC impedence method or linear sweep voltammetry.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104914096A (en) * | 2015-05-07 | 2015-09-16 | 西北农林科技大学 | Clenbuterol detection operating fluid and detection method thereof |
| CN106124716A (en) * | 2016-07-21 | 2016-11-16 | 武汉市农业科学技术研究院农业环境安全检测研究所 | A kind of Clenbuterol method for quick |
| CN106290518A (en) * | 2016-08-23 | 2017-01-04 | 衡阳师范学院 | A kind of molecular imprinting electrochemical sensor for albuterol detection by quantitative and preparation method thereof |
| CN107764880A (en) * | 2017-10-11 | 2018-03-06 | 浙江海洋大学 | A kind of biosensor thin layer electrode of quick detection miocardial infarction protein marker |
-
2014
- 2014-06-27 CN CN201410300939.XA patent/CN104020202A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| PENG MIAO 等: "《ACS Applied Materials & Interfaces》", 《MELAMINE FUNCTIONALIZED SILVER NANOPARTICLES AS THE PROBE FOR ELECTROCHEMICAL SENSING OF CLENBUTEROL》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104914096A (en) * | 2015-05-07 | 2015-09-16 | 西北农林科技大学 | Clenbuterol detection operating fluid and detection method thereof |
| CN104914096B (en) * | 2015-05-07 | 2018-02-16 | 西北农林科技大学 | A kind of Clenbuterol detection working solution and detection method |
| CN106124716A (en) * | 2016-07-21 | 2016-11-16 | 武汉市农业科学技术研究院农业环境安全检测研究所 | A kind of Clenbuterol method for quick |
| CN106290518A (en) * | 2016-08-23 | 2017-01-04 | 衡阳师范学院 | A kind of molecular imprinting electrochemical sensor for albuterol detection by quantitative and preparation method thereof |
| CN107764880A (en) * | 2017-10-11 | 2018-03-06 | 浙江海洋大学 | A kind of biosensor thin layer electrode of quick detection miocardial infarction protein marker |
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Application publication date: 20140903 |