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Immunoaffinity stir bar for adsorbing diethylstilbestrol drug, and preparation method and application thereof

A technology of diethylstilbestrol and immunoaffinity, which is applied in the field of adsorption of diethylstilbestrol-like drugs immunoaffinity stirring rods and its preparation, can solve the problems of poor specificity, environmental pollution, and difficult degradation, and achieve strong specificity, save organic solvents, and analyze The effect of quality improvement

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 disadvantage is its low specificity.
Immunoaffinity SPME technology was first proposed in 2001. Purification of stilbene compounds with Immunoaffinity stir bar has not been reported yet, and there is no commercialized Immunoaffinity stir bar for sale.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Preparation of monoclonal antibody adsorbed to diethylstilbestrol

[0043] Diethylstilbestrol-monocarboxypropyl ether (DES-CP) synthesis: Weigh 55 mg (Diethylstilbestrol, DES) and dissolve it in 2 mL dimethyl sulfoxide (Dimethyl sulfoxide, DMSO), add appropriate amount of anhydrous potassium carbonate and 4-bromobutyl Acetate ethyl ester (Ethyl 4-bromobutyrate), 70 ℃ stirring reaction, thin layer chromatography to monitor the reaction progress. After the reaction was complete, the product was added to pure water, extracted with ethyl acetate, washed with pure water, anhydrous Na 2 SO 4 Dry, blow dry, dissolve in methanol, hydrolyze with NaOH solution (3 mol / L), adjust pH to about 3 with HCl solution (6 mol / L), extract with ethyl acetate, wash with pure water, dry with anhydrous Na2SO4, and blow dry.

[0044] Preparation of diethylstilbestrol-monocarboxypropyl ether-BSA (DES-CP-BSA) immunogen: 40 mg DES-CP was dissolved in 2 mL DMSO, and 14 mg N-hydroxysuccinimide ...

Embodiment 2

[0060] Comparative Test

[0061]

Embodiment 3

[0063] Immunoaffinity stir bars were used for the extraction of stilbenes in water. The blank water sample was passed through a 0.45 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 stilbene compound standard solution was added, vortexed and mixed, and the immunoaffinity Put the glass rod into the water sample after removing the mixing, and vibrate 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 diethylstilbestrol, hexestrol, dienestrol and butestrol could be detected, indic...

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PUM

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Abstract

The invention discloses an immunoaffinity stir bar for adsorbing diethylstilbestrol drug, and a preparation method and an application thereof. The immunoaffinity stir bar for adsorbing diethylstilbestrol drug is composed of solid phase carrier and diethylstilbestrol murine monoclonal antibody coupled with the same, wherein the ratio of the coupling length of the solid phase carrier and coupled diethylstilbestrol DES-6B4 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 toluylene compound 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 for adsorbing diethylstilbestrol drugs, 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 estrogen residues, and relevant food safety departments are also committed to the research of detection methods. Due to the complex composition of biological samples, the easy combination of estrogen and sample components, and the low residual concentration, it has brought great difficulties to the extraction and purification of drugs in the sample. Therefore, the selectivity and sensitivity of the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/543
Inventor 李存姚凯张伟杨琳燕彭涛赵思俊丁巧玲何建丽
Owner TIANJIN AGRICULTURE COLLEGE
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