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ELISA kit for detecting beta-stimulants and detection method thereof

An enzyme-linked immunosorbent reagent and stimulant technology, which is applied in the field of detection and analysis of veterinary drug residues, can solve problems such as complex instruments and equipment, cumbersome processes, unsuitability for on-site monitoring and screening of a large number of samples, and achieve simple pre-treatment process and pre-treatment requirements. low effect

Active Publication Date: 2006-05-03
BEIJING WANGER BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The chemical methods for detection of β-stimulant residues mainly include thin layer chromatography (TLC), gas chromatography (GC), high performance liquid chromatography (HPLC), gas-mass spectrometry (GC / MS), liquid-mass spectrometry (HPLC / MS), etc., due to complex equipment and cumbersome process, it is not suitable for on-site monitoring and screening of a large number of samples

Method used

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  • ELISA kit for detecting beta-stimulants and detection method thereof
  • ELISA kit for detecting beta-stimulants and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] The preparation of embodiment 1 kit components

[0051] 1. Antigen preparation

[0052] a. Synthesis of hapten Using clenbuterol as an intermediate, acylation reaction with succinic anhydride, acylation of the alcohol hydroxyl group on the molecular structure of clenbuterol into a carboxyl spacer arm containing 4 carbons to prepare β-excited agent drug hapten.

[0053] b. Immunogen The β-agonist drug hapten and thyroid protein (BCG) were coupled by the mixed anhydride method (isobutyl chloroformate) to obtain the immunogen.

[0054] The preparation process of the immunogen: Take β-stimulant drug hapten 2g and dissolve it in 20mL, 0.5M sodium hydroxide solution, then take 1.5g carbodiimide and dissolve it in 5mL pure water and add it to the β-stimulant drug Stir and react in the drug hapten solution at room temperature for 2 hours, take 20 g of carrier protein and dissolve it in 75 mL of pH9.6 carbonate buffer solution, then add thyroid protein (BCG) dropwise to the β-...

Embodiment 2

[0072] Embodiment 2 detects the formation of the ELISA kit of β-agonists

[0073] Construct the ELISA kit for detecting β-agonists so that it contains the following components:

[0074] (1) A microtiter plate coated with a conjugate of clenbuterol and human serum albumin (HSA);

[0075] (2) horseradish peroxidase-labeled goat anti-rabbit anti-antibody;

[0076] (3) Clenbuterol standard solution, the concentrations are 0μg / L, 0.1μg / L, 0.3μg / L, 0.9μg / L, 2.7μg / L, 8.1μg / L;

[0077] (4) Substrate chromogenic solution A liquid is hydrogen peroxide, and substrate chromogenic solution B liquid is o-phenylenediamine;

[0078] (5) The concentration of the β-agonist drug rabbit polyclonal antibody working solution is 0.5 μg / L;

[0079] (6) Concentrated washing liquid 0.8% Tween 20 phosphate buffer (0.01M, PH7.4);

[0080] (7) The stop solution is 1mol / L hydrochloric acid;

[0081] (8) The concentrated complex solution is 0.01M phosphate buffer containing 1% casein.

Embodiment 3

[0082] Embodiment 3 detects the formation of the ELISA kit of β-agonist

[0083] Construct the ELISA kit for detecting β-agonists so that it contains the following components:

[0084] (1) A microtiter plate coated with a conjugate of β-agonist drug hapten and human serum albumin (HSA);

[0085] (2) horseradish peroxidase-labeled goat anti-rabbit anti-antibody;

[0086] (3) Clenbuterol standard solution, the concentrations are 0μg / L, 0.1μg / L, 0.3μg / L, 0.9μg / L, 2.7μg / L, 8.1μg / L;

[0087] (4) Substrate chromogenic solution A liquid is carbamide peroxide, and substrate chromogenic solution B liquid is tetramethylbenzidine;

[0088] (5) The concentration of the β-agonist drug rabbit polyclonal antibody working solution is 5.0 μg / L;

[0089] (6) The concentrated washing solution is a phosphate buffered saline solution of 1.2% Tween 20;

[0090] (7) The stop solution is 2mol / L sulfuric acid;

[0091] (8) The concentrated complex solution is 0.05M phosphate buffer containing 1% ...

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Abstract

The invention relates to an enzyme immune agent box for detecting ª‰-excitant drugs, which comprises: enzyme mark plate which coats ª‰-excitant drugs, enzyme mark antibody, clenobuterol standard solution, base material color developing solution, ª‰-excitant drugs antibody working solution, compression cleaning liquid, ending solution and compression twin solution. The invention also discloses a method for applying the detecting method, which comprises: first doing sample front process, then using the agent box to detect, at last analyzing the detected result.

Description

technical field [0001] The invention relates to the technical field of detection and analysis of veterinary drug residues, in particular to an ELISA kit and method for detecting β-stimulants in urine, animal tissues (muscle, liver) and feed. Background technique [0002] Salbutamol (Salbutamol) and Clenbuterol (Clenbuterol) are both β-receptor agonists, which are mainly used clinically to treat bronchial asthma; these drugs can also be used as growth-promoting agents for cattle, sheep, pigs, poultry, etc. It is widely used in animal and poultry production. However, because β-stimulants tend to accumulate residues in animal livers, and can enter the human body through the food chain, seriously endangering human health, so the European Community countries have classified such drugs as banned. Therefore, it is necessary to strengthen the detection of β-agonist residues in animal food. [0003] The chemical methods for detection of β-stimulant residues mainly include thin laye...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/52G01N33/535
Inventor 沈建忠何方洋冯才伟万宇平吴小平冯才茂汪善良李军赵正苗张照亮史为民张素霞丁双阳罗晓琴孙倩
Owner BEIJING WANGER BIOTECH
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