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Method for purifying alpha-zeranol and special immunity affinity chromatographic column therefor

A technology of zearalanol and immunophile, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of excessive waste of organic solvents, cumbersome treatment process, poor purification effect, etc., and achieve improved analysis quality, good purification effect, Simple operation effect

Inactive Publication Date: 2006-07-26
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The pretreatment of these methods utilizes liquid-liquid distribution and conventional SPE column purification and separation, all of which have disadvantages such as cumbersome treatment process, poor purification effect, waste of organic solvents, and long time required to varying degrees.

Method used

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  • Method for purifying alpha-zeranol and special immunity affinity chromatographic column therefor
  • Method for purifying alpha-zeranol and special immunity affinity chromatographic column therefor
  • Method for purifying alpha-zeranol and special immunity affinity chromatographic column therefor

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Embodiment 1, the preparation of the immunochromatographic column of purifying α-zearalenol

[0029] 1. Preparation of α-zearalanol mouse monoclonal antibody

[0030] 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:

[0031] 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.

[0032] 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.

[0033] 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 acetate.

[0034] 4) The pH of the aqueous phase was adjusted...

Embodiment 2

[0060] Example 2, Preparation of the kit of immunochromatographic column coupled with mouse monoclonal antibody and its purification effect on α-zearalanol

[0061] 1. Preparation of a kit for purifying α-zearalanol

[0062] 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 standard α-zearalanol reagents, washing solution, eluent, and reagent bottles of preservation solution, and the IAC column is housed in the hole of the sponge bracket 7 . The immunochromatographic column is the immunochromatographic column of the monoclonal antibody secreted by the monoclonal hybridoma cell line A-1-2 CGMCC No.1604 prepared in Example 1 coupled with α-zearalanol.

[0063] The washing solution is phosphate buffer (0.01M, pH7.4), and the preparation ...

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Abstract

The invention discloses a method for purifyingalpha-gibberella zeae alcohol and the private immunity affinity chromatography thereof. The filling material of the chromatography are alpha-gibberella zeae alcohol and immunity affinity adsorbent which comprises solid carrier and the coupled alpha-gibberella zeae alcohol monoclonal antibody; thealpha-gibberella zeae alcohol monoclonal antibody is made by the coupling object of the alpha-gibberella zeae alcohol and carrier protein as immunogen; The invention has the advantages of having more information and accurate definite quantity.

Description

technical field [0001] The invention relates to a method for purifying α-zearalanol and a special immunoaffinity adsorption chromatographic column thereof. 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. Applying immunoaffinity chromatography (IAC, immunoaffinity chromatography) to residue analysis, its high selectivity and high affinity will undoubtedly simplify the purification process. Combined with GC and HPLC, immunological technology and physical and chemical technology can be complemented in terms of sel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/547
Inventor 沈建忠史为民张伟何方洋王鹤佳
Owner CHINA AGRI UNIV
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