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
View PDF14 Cites 2 Cited by
  • 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 disadvantage is its low specificity.
Immunoaffinity SPME technology was first proposed in 2001, and the purification of CAP with Immunoaffinity stir bar has not been reported yet, and there is no commercialized Immunoaffinity stir bar for sale

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • 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
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Preparation of chloramphenicol mouse monoclonal antibody

[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 (N,N-Dimethylformamide, DMF) and 1 mL of 1,4 - Dioxane (1,4-dioxane) mixed solution, then add 5 mL of isobutyl chloroformate (Isobutyl chloroformate), 4 ℃ low-speed stirring for 30 min, dropwise added to the BSA solution (50 mg of BSA dissolved In 1 mL of 0.02 mol / L PBS), stirred at 4 °C for 4 h at low speed, and the PBS solution (0.01 mol / L) was stirred and dialyzed at 4 °C for 5 days.

[0044] The coated antigen CAP-OVA was synthesized by the diazonium method. Take 80 mg of CAP, 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 water bath at 70 °C for 50 min. Add NaNO dropwise at 4°C and pH 1-2 2 (0.5 mol / L), and stir in the dark until the starch pot...

Embodiment 2

[0059] Comparative Test

[0060]

Embodiment 3

[0062] Immunoaffinity stir bar was used for the extraction of chloramphenicol in water. The blank water sample was passed through a 0.45 μm water filter membrane to remove impurities, and 5 mL of the filtered water sample was accurately weighed, placed in a 10 mL centrifuge tube, an appropriate amount of chloramphenicol standard solution was added, vortexed, and the immunophile Put it and the glass rod into the water sample after removing the mixing, and shake it on the shaker for 30 min. Take the oscillated glass rod out of the centrifuge tube, wash it with ultrapure water, put it into a 2 mL centrifuge tube filled with 700 μL of methanol, shake it on a shaker for 20 min to elute, dry it with nitrogen, and use the mobile phase Reconstitution, filtration with 0.22 μm microporous membrane, separation and detection by high performance liquid chromatography-mass spectrometry. The results showed that chloramphenicol and succinic chloramphenicol could be detected, indicating that...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
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
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap