Furacillin metabolite SEM fast detecting reagent kit and uses thereof

A detection kit, nitrofurazone technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of being unsuitable for rapid detection of a large number of samples, high instrument requirements, and cumbersome detection.

Inactive Publication Date: 2009-01-21
深圳市绿诗源生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The most commonly used methods for the detection of nitrofurazone metabolites are LC-UV, LC-MS and LC-MS / MS, but these methods are relatively cumbersome and require high equipment requirements, and are not suitable for rapid detection of a large number of samples

Method used

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  • Furacillin metabolite SEM fast detecting reagent kit and uses thereof
  • Furacillin metabolite SEM fast detecting reagent kit and uses thereof
  • Furacillin metabolite SEM fast detecting reagent kit and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0100] Example 1 Preparation of ELISA kit components for detecting nitrofurazone metabolites

[0101] 1. Antigen Synthesis

[0102] a. Synthesis of Coating Progen

[0103] The nitrofurazone metabolite is synthesized by a derivative method to synthesize a nitrofurazone metabolite derivative hapten, and then the hapten is obtained through diazotization reaction and bovine gamma globulin carrier protein coupling by an active ester method.

[0104] b. Synthesis of Immunogen

[0105] The nitrofurazone metabolite is synthesized by a derivative method to synthesize a nitrofurazone metabolite derivative hapten, and then the hapten is obtained through diazotization reaction and keyhole limpet hemocyanin carrier protein coupling by an active ester method.

[0106] 2. Preparation of mouse monoclonal antibody against nitrofurazone metabolite derivatives

[0107] a. Animal immunity

[0108] Balb / c mice were used as immunized animals, and the conjugated hapten of nitrofurazone metabolit...

Embodiment 2

[0119] Example 2 The formation of an enzyme-linked immunosorbent assay kit for detecting nitrofurazone metabolites

[0120] An enzyme-linked immunosorbent assay kit for detecting nitrofurazone metabolites was set up to include the following components:

[0121] (1) Enzyme-linked plates coated with nitrofurazone metabolite derivative antigens;

[0122] (2) Goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0123] (3) Rat monoclonal antibody to nitrofurazone metabolite derivatives;

[0124] (4) 6 bottles of nitrofurazone metabolite standard solution, the concentrations were 0 μg / L, 0.05 μg / L, 0.15 μg / L, 0.45 μg / L, 1.35 μg / L, 4.05 μg / L;

[0125] (5) The substrate chromogenic solution A liquid is hydrogen peroxide, and the substrate chromogenic liquid B liquid is o-phenylenediamine;

[0126] (6) The stop solution is 2mol / L. sulfuric acid buffer;

[0127] (7) The concentrated washing solution is pH 7.4, containing 0.1%-0.5% Tween 80, 0.5% sodium azide phospha...

Embodiment 3

[0130] Embodiment 3 detects the formation of the ELISA kit of nitrofurazone metabolite

[0131] An enzyme-linked immunosorbent assay kit for detecting nitrofurazone metabolites was set up to include the following components:

[0132] (1) Enzyme-linked plate coated with goat anti-rabbit anti-antibody;

[0133] (2) Furacilin metabolite derivative antigen labeled with alkaline phosphatase;

[0134] (3) Rabbit polyclonal antibody against nitrofurazone metabolite derivatives;

[0135] (4) 6 bottles of nitrofurazone metabolite derivative standard solution, the concentrations are 0 μg / L, 0.05 μg / L, 0.15 μg / L, 0.45 μg / L, 1.35 μg / L, 4.05 μg / L;

[0136] (5) Substrate chromogenic solution p-nitrophosphate buffer;

[0137] (6) The stop solution is 2mol / L sodium hydroxide buffer solution;

[0138] (7) The concentrated washing liquid is pH 7.4, containing 0.8% Tween 20 and 0.1% sodium azide (NaN 3 ) phosphate buffered saline solution with preservatives;

[0139] (8) The concentrated c...

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Abstract

The invention provides an enzyme linked immunosorbent kit for detecting furacilin metabolites, which comprises an enzyme-linked plate which is coated with a coating source, an enzyme marker, a furacilin metabolite derivant specific antibody, furacilin metabolite derivant standardized product solution, substrate coloured solution, stopping solution, concentrated cleaning mixture and concentrated complex solution. Furacilin metabolite derivant which is coated with antigen is got through coupling furacilin metabolite derivant hapten and bovine gamma globulin, and the antibody is a polyclonal antibody or a monoclonal antibody. The invention also provides a method for detecting furacilin metabolites in animal derived food, which comprises the following steps: preprocessing samples, detecting by agent in a kit, and analyzing detection results. The invention aims at providing an enzyme linked immunosorbent kit and a detection method for detecting furacilin metabolites in animal derived food, which has simple operation and low cost, and is suitable for screening a great amount of sample copies.

Description

technical field [0001] The invention relates to a technique for detecting and analyzing veterinary drug residues in animal-derived food, in particular to an enzyme-linked immunosorbent reagent and a method for detecting nitrofurazone. Background technique [0002] Nitrofuran drugs have been widely used as growth-promoting additives for poultry, aquatic products and pigs because of their excellent antibacterial and pharmacokinetic properties. However, in the course of long-term experimental research, it was found that both nitrofuran drugs and metabolites (SEM) can cause cancer and gene mutations in experimental animals, which is why such drugs are prohibited from being used in treatment and feed. Disabled in 1995. [0003] Since nitrofuran drugs can be metabolized quickly in the body, and the metabolites combined in tissues can persist for a long period of time, it is often necessary to analyze the metabolized products when analyzing the residues of such drugs. The managem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/577
Inventor 聂继斌唐俊杨宏温俊梅齐欣吴青李成杨宗繁
Owner 深圳市绿诗源生物技术有限公司
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