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ELISA kit for detecting diazepam residue and detection method thereof

A test kit and stable technology, applied in the field of enzyme-linked immunosorbent assay kits, can solve the problems of unsuitable screening, high cost, cumbersome processing and measurement operations, etc.

Inactive Publication Date: 2006-05-03
贵州勤邦食品安全科学技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High performance liquid chromatography has good repeatability, but sample pretreatment and measurement operations are cumbersome and costly, and are not suitable for screening a large number of samples

Method used

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  • ELISA kit for detecting diazepam residue and detection method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1 Component preparation of an enzyme-linked immunosorbent assay kit for detecting diazepam residues

[0077] 1. Antigen Synthesis

[0078] a. Synthesis of Coating Progen

[0079] Diazepam and carboxymethylhydroxylamine are condensed into a hapten, and a spacer arm containing 2 carbons is connected to diazepam, which highlights all the characteristic structures of diazepam. The hapten and the human serum albumin carrier protein are coupled by a water-soluble carbodiimide method to obtain the coating original.

[0080] b. Synthesis of Immunogen

[0081] Diazepam and carboxymethylhydroxylamine are condensed into a hapten, and a spacer arm containing 2 carbons is connected to diazepam, which highlights all the characteristic structures of diazepam. The hapten and the thyroid protein carrier protein are coupled by the water-soluble carbodiimide method to obtain the immunogen.

[0082] 2. Preparation of stable mouse monoclonal antibody

[0083] a. Animal immunity...

Embodiment 2

[0093] Embodiment 2 detects the formation of stable ELISA kit

[0094] Set up the enzyme-linked immunosorbent assay kit for detecting stability, so that it includes the following components:

[0095] (1) Enzyme-linked plate coated with diazepam antigen;

[0096] (2) Goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0097] (3) stable mouse monoclonal antibody;

[0098] (4) 6 bottles of diazepam standard solution, the concentrations are 0μg / L, 0.1μg / L, 0.2μg / L, 0.4μg / L, 0.8μg / L, 1.6μg / L;

[0099] (5) Chromogenic solution A liquid is hydrogen peroxide, and chromogenic liquid B liquid is o-phenylenediamine;

[0100] (6) The stop solution is 1mol / L sulfuric acid buffer;

[0101] (7) The concentrated washing solution is pH 7.4, containing 0.8% to 1.2% Tween 20 and 1‰ sodium azide (NaN 3 ) phosphate buffered saline solution with preservatives;

[0102] (8) The concentrated complex solution is a phosphate buffer containing 5‰N,N'-dimethylformamide (DMF).

Embodiment 3

[0103] Embodiment 3 detects the formation of stable ELISA kit

[0104] Set up the enzyme-linked immunosorbent assay kit for detecting stability, so that it includes the following components:

[0105] (1) Enzyme-linked plate coated with goat anti-rabbit anti-antibody;

[0106] (2) Diazepam antigen labeled with alkaline phosphatase;

[0107] (3) Diazepam rabbit polyclonal antibody;

[0108](4) 6 bottles of diazepam standard solution, the concentrations are 0μg / L, 0.1μg / L, 0.2μg / L, 0.4μg / L, 0.8μg / L, 1.6μg / L;

[0109] (5) The substrate solution is p-nitrophosphate buffer;

[0110] (6) The stop solution is sodium hydroxide of 2mol / L;

[0111] (7) The concentrated washing solution is pH 7.4, containing 0.8% to 1.2% Tween 20 and 1‰ sodium azide (NaN 3 ) phosphate buffered saline solution with preservatives;

[0112] (8) The concentrated complex solution is a phosphate buffer containing 5‰N,N'-dimethylformamide (DMF).

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Abstract

The invention provides an enzyme immune agent box for detecting atilen residual quantity of animal foodstuff which comprises: enzyme mark plate which coats atilen antigen or antibody, enzyme mark material, atilen peculiar antibody, atilen standard solution, color developing 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 an ELISA kit and method for detecting diazepam residues in animal-derived foods. Background technique [0002] Diazepam, as a sedative, is widely used in livestock and poultry, which can reduce the amount of exercise of livestock and poultry animals, increase the weight of animals and ensure that livestock and poultry will not produce stress reactions during long-distance transportation. However, a large amount of diazepam taken by animals will lead to drug residues in food and be absorbed by the human body. Large doses can cause central nervous system depression, respiratory depression and other toxic manifestations. Long-term use of this type of drug can lead to dependence or addiction, and sudden withdrawal of the drug can cause withdrawal reactions, manifested as depression, agitation, and insomnia. [0003] At present, the method used for the detection of diazepam residues is mainly high performance liquid chromatography....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/52G01N33/535G01N33/543
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 贵州勤邦食品安全科学技术有限公司
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