Immunoaffinity stir bar for adsorbing chloramphenicol, and preparation method and application thereof

A technology of chloramphenicol and immunophile, which is applied in the field of food safety testing, can solve the problems of low specificity, non-commercialization, environmental pollution, etc., and achieve the effects of strong specificity, lower analysis cost and saving organic solvents

Inactive Publication Date: 2015-06-10
TIANJIN AGRICULTURE COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The LLE process needs to use a large amount of organic solvents, and some organic solvents are stable, not easy to degrade, pollute the environment, and the operation is cumbersome
SPE is easy to operate and saves time, but its main disad

Method used

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  • Immunoaffinity stir bar for adsorbing chloramphenicol, and preparation method and application thereof
  • Immunoaffinity stir bar for adsorbing chloramphenicol, and preparation method and application thereof

Examples

Experimental program
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Example Embodiment

[0041] Example 1

[0042] 1. Preparation of mouse monoclonal antibody to chloramphenicol

[0043] The CAP-BSA immune antigen was synthesized by the mixed acid anhydride method, and 20 mg of chloramphenicol (Chloramphenicol, CAP) was dissolved in 1 mL of N,N-Dimethylformamide (DMF) and 1 mL of 1,4 -Dioxane (1,4-dioxane) mixed solution, then add 5 mL of Isobutyl chloroformate, stir at 4 ℃ for 30 min at low speed, and add dropwise to the BSA solution (50 mg of BSA is dissolved In 1 mL of 0.02 mol / L PBS, stir at 4 ℃ for 4 h at low speed, and PBS solution (0.01 mol / L) was stirred and dialyzed at 4 ℃ for 5 d.

[0044] To synthesize coated antigen CAP-OVA by diazonium method, take 80 mg of CAP and dissolve it in 1.5 mL of absolute ethanol, add 12 mL of hydrochloric acid (1 mol / L), add 65 mg of zinc powder, and heat in a 70 ℃ water bath for 50 min. Add NaNO dropwise at 4℃ and pH 1-2 2 (0.5 mol / L), avoid light and stir until the starch potassium iodide test paper is grayish blue. While stir...

Example Embodiment

[0058] Example 2

[0059] Comparative Test

[0060]

Example Embodiment

[0061] Example 3

[0062] The immunoaffinity stirring rod is used for the extraction of chloramphenicol in water. The blank water sample is passed through a 0.45μm water filter membrane to remove impurities. Weigh accurately 5 mL of the filtered water sample and place it in a 10 mL centrifuge tube. Add an appropriate amount of chloramphenicol standard solution, vortex to mix, Put the glass rod into the water sample after removing and mixing, and oscillate on a shaker for 30 min. Take out the shaken glass rod from the centrifuge tube, wash it with ultrapure water, put it into a 2 mL centrifuge tube containing 700 μL of methanol, shake on a shaker for 20 minutes for elution, blow dry with nitrogen, and move the mobile phase Reconstituted, filtered with 0.22 μm microporous membrane, and separated and detected by high performance liquid chromatography-mass spectrometry. The results show that chloramphenicol and succinic chloramphenicol can be detected, indicating that two chloramp...

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Abstract

The invention discloses an immunoaffinity stir bar for adsorbing chloramphenicol, and a preparation method and an application thereof. The immunoaffinity stir bar for adsorbing chloramphenicol is composed of solid phase carrier and chloramphenicol murine monoclonal antibody coupled with the same, wherein the ratio of the coupling length of the solid phase carrier and coupled chloramphenicol murine monoclonal antibody to the weight thereof is 1 cm: 2 mg. Through combining with a chromatography, an extracting method can effectively separate and condense the ingredients to be detected in a sample, the chloramphenicol content detection is facilitated, the disadvantages of less information amount, low quantification precision, low physico-chemical method selectivity and the like due to using the simplex immunoassay to directly detect the sample are overcome, and the analysis mechanism complementarity between the immunological technique and conventional physical and chemical technology is reflected.

Description

technical field [0001] The invention belongs to the technical field of food safety detection, and relates to an immunoaffinity stirring rod and a preparation method thereof, in particular to an immunoaffinity stirring rod adsorbing chloramphenicol, a preparation method and application thereof. Background technique [0002] With the development of my country's economy and the improvement of people's living standards, the demand for various animal-derived foods is gradually increasing, and food safety issues have naturally become the focus of everyone's attention. In recent years, there have been many reports about the harm to human health caused by veterinary drug residues, and relevant food safety departments are also committed to the research of detection methods. Due to the complex components of biological samples, the easy combination of drugs and sample components, and the low concentration of residual drugs, it has brought great difficulties to the extraction and purifi...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/54353G01N33/531G01N33/552
Inventor 李存姚凯彭涛张伟杨琳燕赵思俊何建丽王翠翠
Owner TIANJIN AGRICULTURE COLLEGE
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