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Quick test method fro detecting clenbuterol hydrochloride in animal derived food

A technology of clenbuterol hydrochloride and animal food, which is applied in the field of detection, can solve the problems of insensitivity, insufficient specificity, and prolonged time, and achieve the effects of simple and time-saving operation, improved sensitivity, and broad application prospects

Inactive Publication Date: 2006-05-03
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

But the defect of this method is: when the antibody on the gold label pad is combined with the clenbuterol-protein conjugate on the nitrocellulose membrane, it is difficult for the clenbuterol in the sample to make the bound gold in a short time. The clenbuterol antibody is lost from the test line, so the test is not sensitive and takes a long time
However, this technology still uses the traditional colloidal gold immunochromatography technology, the detection time is long, up to more than 20 minutes, the detection limit cannot reach 0.3ppb (μg / L), and the specificity of detection is not strong enough
In the patents currently applied for, the detection limit of clenbuterol hydrochloride with colloidal gold immunoassay strips is generally above 0.09ppb (ng / ml), so it cannot meet the needs of rapid and ultra-trace detection

Method used

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  • Quick test method fro detecting clenbuterol hydrochloride in animal derived food
  • Quick test method fro detecting clenbuterol hydrochloride in animal derived food

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Embodiment

[0033] ①Clenbuterol-glucose oxidase conjugate was synthesized, and the enzyme activity and binding ratio of the conjugate were identified. When identifying enzyme activity, dissolve 3.5mg horseradish peroxidase and 3.5mg 4-aminoantipyridine in 20ml 0.2mol / L pH7.0 phosphate buffer and mix evenly, add 1ml 3% phenol solution to prepare into solution A, and solution B is 6.5% dextrose aqueous solution. During the measurement, keep the mixture of 1.5ml solution A and 1.5ml solution B at 25°C, add the synthesized and purified clenbuterol-glucose oxidase conjugate, and measure A 500 Changes, and calculate the activity of the enzyme. When identifying the binding ratio, prepare clenbuterol and clenbuterol-glucose oxidase conjugates to 200μg / ml and 0.4mg / ml respectively, and then perform UV scanning, according to OD 280 The respective molar absorptivity ε was calculated, and the binding ratio between clenbuterol and glucose oxygenase was determined, and the binding ratio=[ε280(conjuga...

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Abstract

The invention relates to a fast test method for detecting the clenobuterol hydrochloride of animal foodstuff. Firstly syntheses marks the clenobuterol coupled material; then it fixes the horse radish perxidase and the anti-clenobuterol monoclonal antibody on the cellulose nitrate film; then dispositions the cellulose film coated the antibody and the enzyme inside the one time polyvinyl fluoride or the reaction hole which is made by glass material after sealed and dried by the skimmed milk powder and uses the polytef film to seal it; then it prepares the color development bottom material solution and assemblies the clenobuterol fast reacting plate; it mixes the tested sample and the enzyme mark clenobuterol coupled material and adds them on the antibody film of the reaction hole of the reaction plate; it washes it after reacting at indoor temperature and adds bottom material color development solution on the reacting hole to observe the result and then quotes the clenobuterol content by the color development degree.

Description

technical field [0001] The invention relates to a method in the technical field of detection, in particular to a rapid test method for detecting clenbuterol hydrochloride in animal food. Background technique [0002] At home and abroad, the methods involved in the detection of clenbuterol hydrochloride mainly include chromatography, capillary electrophoresis, immunoassay (radioimmunoassay, enzyme immunoassay) and electrochemical detection technology. The most common technique used for rapid detection or rapid semi-quantitative detection is the immunocolloidal gold method. The clenbuterol in the sample and the clenbuterol-protein conjugate immobilized on the nitrocellulose membrane compete to bind the gold-labeled antibody diffused from the sample pad, and the analyte is determined by detecting the color depth of the line content. But the defect of this method is: when the antibody on the gold label pad is combined with the clenbuterol-protein conjugate on the nitrocellulos...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/78G01N33/531G01N33/543
Inventor 柴春彦刘国艳
Owner SHANGHAI JIAO TONG UNIV
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