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Preparation method and application of unmarked immunosensor for rapidly detecting clenbuterol

An immune sensor and clenbuterol technology, applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve the problems of difficult operation, expensive instruments, and low false positive rate, and achieve enhanced adsorption and immobilization, high selectivity detection, The effect of improving sensitivity

Inactive Publication Date: 2013-04-24
UNIV OF JINAN
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Problems solved by technology

Although these methods have certain sensitivity, there are the following problems, which limit the promotion of these methods in practical applications:
[0008]1. High performance liquid chromatography (HPLC): The advantage is that it is suitable for detecting thermally unstable and highly polar β-agonists and their metabolites, with good specificity , strong selectivity, high detection accuracy, can be combined with mass spectrometry (MS) and other systems to realize the automation of the analysis process, the HPLC method has been used as a semi-confirmative method for the detection of clenbuterol residues in my country, the minimum detection limit range It is 1 ~ 15 ng·g-1; the disadvantage is that the sample processing time is long, the detection process is cumbersome, difficult to operate, and the instrument is expensive, which is limited in practical application
[0009]2. Gas Chromatography-Mass Spectrometry (GC-MS): The advantage is that by combining the high-efficiency and rapid separation effect of chromatography with the high-sensitivity qualitative analysis of mass spectrometry, it can Qualitative and quantitative analysis of a specific residue in the presence of multiple residues. Compared with the HPLC method, this method has higher detection sensitivity and lower false positive rate. In my country, GC-MS method is used as the detection gram The confirmatory method of Renbutro; the disadvantage is that the equipment is complicated, the operation process is cumbersome, the operation skills of the inspectors are very high, and it is not suitable for on-site and rapid detection
[0010]3. Enzyme-linked immunoassay (ELISA): The advantage is that by using the immunological antigen-antibody specific binding and the high-efficiency catalysis of the enzyme, it can be detected by visual inspection or colorimetry. Detect the content of clenbuterol in the sample, so the method has strong selectivity and good specificity; the disadvantage is that there are many steps and the detection error is large

Method used

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  • Preparation method and application of unmarked immunosensor for rapidly detecting clenbuterol
  • Preparation method and application of unmarked immunosensor for rapidly detecting clenbuterol
  • Preparation method and application of unmarked immunosensor for rapidly detecting clenbuterol

Examples

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Effect test

Embodiment 1

[0045] Example 1 A preparation method of a label-free immunosensor for rapid detection of clenbuterol:

[0046] (1) Treatment of the glassy carbon electrode: the glassy carbon electrode with a diameter of 4 mm was polished with 1.0, 0.3 and 0.05 μm aluminum oxide polishing powder in sequence, ultrasonically cleaned with ethanol, and then rinsed with ultrapure water to make the glassy carbon The electrode surface is mirror-like.

[0047] (2) Au-Ag 2 Preparation of S core-shell nanocomposites:

[0048] 1) 1 mmol·L in 250 mL -1 Add 60 mL of 10 mmol L to the chloroauric acid solution -1 Freezing point-sodium borohydride solution, after stirring for 1 hour, dilute 10 times to prepare nano-Au seeds;

[0049] 2) 1 mmol·L in 150 mL -1 Add 0.1 mol·L dropwise to the chloroauric acid solution -1 Ascorbic acid until colorless, then add 5 drops of Au seeds in step 1), and let it stand for 2 h to prepare Au nanoparticles;

[0050] 3) After centrifuging the Au nanoparticles prepare...

Embodiment 2

[0060] Example 2 A preparation method of a label-free immunosensor for rapid detection of clenbuterol:

[0061] (1) Treatment of the glassy carbon electrode: same as in Example 1.

[0062] (2) Au-Ag 2 Preparation of S core-shell nanocomposites:

[0063] 1) In 250 mL 1 mmol·L -1 Add 60 mL of 10 mmol L to the chloroauric acid solution -1 Freezing point-sodium borohydride solution, after stirring for 1 hour, dilute 10 times to prepare nano-Au seeds;

[0064]2) 1 mmol·L in 150 mL -1 Add 0.1 mol·L dropwise to the chloroauric acid solution -1 Ascorbic acid until colorless, then add 5 drops of Au seeds in step 1), and let it stand for 2 h to prepare Au nanoparticles;

[0065] 3) After centrifuging the Au nanoparticles prepared in step 2), redisperse them in 300 mL of 0.1 mol L -1 To the CTAB, add 30 mL 0.1 mol·L successively -1 Ascorbic acid, 5 mL 0.01 mol L -1 AgNO 3 , 30 mL 0.1 mol·L -1 NaOH aqueous solution, shaking for 1 min, and standing for 1 h, the Au-Ag nanop...

Embodiment 3

[0072] Example 3 A preparation method for rapid detection of clenbuterol label-free immunosensor, such as figure 1 shown

[0073] (1) Treatment of the glassy carbon electrode: same as in Example 1.

[0074] (2) Au-Ag 2 Preparation of S core-shell nanocomposites:

[0075] 1) 1 mmol·L in 250 mL -1 Add 60 mL of 10 mmol L to the chloroauric acid solution -1 Freezing point-sodium borohydride solution, after stirring for 1 hour, dilute 10 times to prepare nano-Au seeds;

[0076] 2) 1 mmol·L in 150 mL -1 Add 0.1 mol·L dropwise to the chloroauric acid solution -1 Ascorbic acid until colorless, then add 5 drops of Au seeds in step 1), and let it stand for 2 h to prepare Au nanoparticles;

[0077] 3) After centrifuging the Au nanoparticles prepared in step 2), redisperse them in 300 mL of 0.1 mol L -1 To the CTAB, add 30 mL 0.1 mol·L successively -1 Ascorbic acid, 7 mL 0.01 mol L -1 AgNO 3 , 30 mL 0.1 mol·L -1 NaOH aqueous solution, shaking for 1 min, and standing for...

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Abstract

The invention relates to a preparation method and application of an unmarked immunosensor for rapidly detecting clenbuterol. Clenbuterol means a medicine of a beta-stimulant type instead of a specific medicine, and has the functions of prompting lean meat growth and suppressing fat meat growth as well as extremely serious side effects, and when a large amount of clenbuterol is taken, the cardiovascular system of people can be damaged and severe neurological symptoms can be caused. According to the invention, a gold-silver-sulfide nano composite material modified electrode with a core-shell structure which combines the excellent characteristics of nano gold and nano silver sulfide is adopted for preparing the unmarked immunosensor, and a clenbuterol antibody is well immobilized, so that the unmarked immunosensor is low in cost, high in sensitivity, good in specificity, rapid in detection and simple to prepare.

Description

technical field [0001] The invention relates to a preparation method and application of a label-free immunosensor for rapid detection of clenbuterol. Specifically based on gold-silver sulfide (Au-Ag 2 S) A label-free electrochemical immunosensor for rapid detection of clenbuterol constructed from core-shell nanocomposites, which belongs to the technical field of new nano functional materials and biosensors. Background technique [0002] Electrochemical immunosensors are widely used in clinical diagnosis, environmental analysis, drug analysis, food safety detection and other fields due to their high sensitivity, good specificity, and easy operation. The most critical technology for the preparation of electrochemical immunosensors with superior performance is the effective immobilization of immune molecules and the improvement of sensitivity, reproducibility, and persistence. [0003] Gold (Au) nanoparticles have been widely used in the research of immunosensors due to their...

Claims

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Application Information

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IPC IPC(8): G01N33/551G01N27/327
Inventor 张勇魏琴吴丹李贺杜斌王志玲马洪敏庞雪辉李慧芝罗川南范大伟其他发明人请求不公开姓名
Owner UNIV OF JINAN
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