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Method of purifying alpha corn gibberellol and its metabolic substance and immune affinity chromatographic column

A technology for zearalenol and zearalenol, which is applied in the field of purifying alpha-zearalenol and its metabolites, can solve the problems of excessive waste of organic solvents, cumbersome treatment process, poor purification effect, etc. The effect of simplification, good purification, and improved analysis quality

Inactive Publication Date: 2006-09-13
CHINA AGRI UNIV
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  • Application Information

AI Technical Summary

Problems solved by technology

The pretreatment of these methods mostly utilizes liquid-liquid distribution, and conventional SPE column purification and separation have disadvantages such as cumbersome treatment process, poor purification effect, waste of organic solvents, and long time required to varying degrees.
Immunoaffinity technology is a new technology applied in the field of analysis in the 1990s, but the use of immunoaffinity columns to simultaneously purify α-zearalanol (α-Zearalanol, ZER), β-zearalanol (TAL) in the matrix , zearalenone (ZEN) and α-zearalenol (α-ZOL) have not been reported, and there is no commercial IAC column for sale

Method used

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  • Method of purifying alpha corn gibberellol and its metabolic substance and immune affinity chromatographic column
  • Method of purifying alpha corn gibberellol and its metabolic substance and immune affinity chromatographic column
  • Method of purifying alpha corn gibberellol and its metabolic substance and immune affinity chromatographic column

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Embodiment 1

[0026] Embodiment 1, the preparation of the immunochromatographic column of purifying α-zearalenol and / or β-zearalenol and / or zearalenol and / or α-zearalenol

[0027] 1. Preparation of α-zearalanol mouse monoclonal antibody

[0028] Synthesis of hapten: react α-zearalenol with succinic anhydride to obtain ZER-7-hemisuccinate, which is α-zearalenol hapten. The specific steps are as follows:

[0029] 1) Take 520 mg of α-zearalanol and 480 mg of succinic anhydride, dissolve them in 20 ml of pyridine, and heat to reflux for 4 hours under a nitrogen atmosphere.

[0030] 2) The reaction product of step 1) was evaporated under reduced pressure, the residue was dissolved in ethyl acetate, then washed 3 times with 0.5 mol / L sulfuric acid, and the aqueous phase was discarded to obtain an organic phase.

[0031] 3) The organic phase obtained in step 2) was extracted 3 times with a saturated sodium carbonate solution, and the aqueous phase was retained, and washed 3 times with ethyl acet...

Embodiment 2

[0058] Example 2, Preparation of the kit of immunochromatographic column coupled with mouse monoclonal antibody and its purification effect on α-zearalanol

[0059] 1. Preparation of a kit for purifying α-zearalenol and / or β-zearalenol and / or zearalenol and / or α-zearalenol

[0060] It is mainly composed of a box body, an immunochromatographic column (IAC column), a standard α-zearalanol reagent, a washing solution, an eluent, a preservation solution, and a sponge bracket. The sponge bracket is provided with holes and grooves. In the groove of the sponge bracket, there are reagent bottles containing standard α-zearalanol reagent, washing solution, eluent, and preservation solution, and the hole of the sponge bracket is equipped with an IAC column. Wherein the immunochromatographic column prepared in Example 1 is coupled with monoclonal hybridoma cells with cross-reactivity to α-zearalenol, β-zearalenol, zearalenol and α-zearalenol Immunochromatographic column of monoclonal ant...

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Abstract

A method for purifying alpha-zearalol and / or beta-zearalol and / or zearalone and / or alpha-zearalenol by the special immunoaffinity chromatographic column carrying immunoaffinity adsorbent is disclosed. Said adsorbent is composed of solid carrier and its coupled alpha-zearalol monoclonal antibody. It features that the chromatography is used to measure the contents of alpha-zearalol and its metabolites, having high correctness.

Description

technical field [0001] The invention relates to a method for purifying α-zearalenol and its metabolites and its special immunoaffinity chromatographic column, in particular to a method for purifying α-zearalenol and / or β-zearalenol and / or zearalenol Method for mycomycin and / or α-zearalenol and its dedicated immunoaffinity column. Background technique [0002] With the development of life sciences, people have become more and more interested in the substances and their changes in living organisms, and the analysis of biological samples has become a necessary means to explore and discover the mysteries of life. Due to the complex composition of biological samples, the concentration of analytes is low, and most of the sample volumes are small, which puts forward higher requirements for the selectivity and sensitivity of the analytical method. Immunoaffinity chromatography (IAC, immunoaffinity chromatography) is applied to the analysis of veterinary drug residues, and its high ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/281
Inventor 沈建忠史为民张伟何方洋王鹤佳
Owner CHINA AGRI UNIV
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