Beta analeptic clenbuterol immune affinity column, its preparing method and use

An immunoaffinity and affinity technology, applied in the field of food safety, detection and analysis of prohibited agricultural and veterinary drugs

Inactive Publication Date: 2007-06-13
北京中一茶文化发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Immunoaffinity column is a technology that was applied in the field of analysis in the 1990s, but the extraction of clenbuterol in samples by immunoaffinity column has not been reported yet

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Preparation of Anti-Clenbuterol Polyclonal Antibody

[0036] 1.1 Preparation of anti-clenbuterol polyclonal antibody

[0037] Refer to the conjugate synthesis method in the prior art (Itaru Yamamoto, Kohji Iwata Enzyme immunoassay forclenbuterol, an β2-adrenergic stimulant Journal of Immunoassay, 3(2), 155-171 (1982)) to synthesize clenbuterol-carrier protein Conjugate. The specific operation is as follows. Weigh 31.3 mg of clenbuterol hydrochloride (about 0.1 mmol), dissolve it in 4ml of deionized water, adjust the pH to 1.5-2 with 1M hydrochloric acid, and add 0.1M NaNO dropwise under dark conditions at 4°C. 2 , Keep stirring and react for 30 minutes, test with KI starch test paper, stop the reaction until the test paper turns grayish blue, and remove excess NaNO with ammonium sulfamate 2 , Get diazotized Clenbuterol.

[0038] Take an appropriate amount of diazotized clenbuterol hydrochloride solution and add it dropwise to 10ml, 0.1M pH 7.5 ice-cold carrier pro...

Embodiment 2

[0052] Example 2 Preparation of Clenbuterol immunoaffinity column using Sepherose 4B as matrix

[0053] This example is the preparation of Clenbuterol immunoaffinity column using Sepherose 4B as matrix

[0054] Take Seperose 4B (100-120 mesh) wet weight 4g (6ml), use 0.1M pH9.5 Na 2 CO 3 The solution is soaked for 4 hours. Weigh 0.8g cyanogen bromide in a fume hood and dissolve it in 1.2ml Na 2 CO 3 (2g solution 3ml buffer solution), electromagnetic stirring for 10 minutes. Under electromagnetic stirring in an ice bath, add the cyanogen bromide solution to Seperose 4B within 2 minutes. Add 2N NaOH dropwise to keep the pH at 11 and react in an ice bath for 10 minutes.

[0055] Pour the reacted Sepherose 4B into the Buchner funnel, use pre-cooled Na 2 CO 3 The solution is washed at 4-10°C within 2-3 minutes, and 25 times the volume of the liquid is pumped to prepare the cross-linked protein.

[0056] At 4°C, the antibody (polyclonal antibody or monoclonal antibody) solution prepared...

Embodiment 3

[0061] Example 3 Preparation of Clenbuterol immunoaffinity column using dextran as matrix

[0062] This example is the preparation of Clenbuterol immunoaffinity column using dextran as the matrix

[0063] Take dextran (100-120 mesh) wet weight 4g (6ml), use 0.1M pH9.5 Na 2 CO 3 The solution is soaked for 4 hours. Weigh 0.8g sodium periodate and dissolve in 1.2ml Na in a fume hood 2 CO 3 (2g solution 3ml buffer solution), electromagnetic stirring for 10 minutes. Under ice-bath electromagnetic stirring, add sodium periodate solution to dextran, complete within 2 minutes, add 2N NaOH dropwise to keep the pH at 11, and react in ice-bath for 10 minutes.

[0064] Pour the reacted dextran into the Buchner funnel and use pre-cooled Na 2 CO 3 The solution is washed at 4-10°C within 2-3 minutes, and 25 times the volume of the liquid is pumped to prepare the cross-linked protein.

[0065] At 4°C, the antibody (polyclonal antibody or monoclonal antibody) solution prepared in Example 1 was mix...

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PUM

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Abstract

The invention discloses a method to distill beta2-excitant immune avidity column that could be used to distill beta2-exitant in forage and cattle. It includes beta2-excitant specificity antibody, coupling with beta2-exitant specificity antibody Sepharose4B, and plastic column, buffering liquid, eluate and eluent. The invention is convenient to use, has high specificity, and could rapidly distil beta2-exitant in forage and cattle.

Description

Technical field [0001] The invention belongs to the technical field of detection and analysis of illegal agricultural and veterinary drugs in the field of food safety. Specifically, the present invention relates to a β-agonist-clenbuterol immunoaffinity chromatography column and its preparation method and its use. Background of the invention [0002] Clenbuterol, commonly known as Clenbuterol, is a β2-receptor stimulant. It was once used as a drug to treat asthma. It is currently illegally used in livestock production. It is not easily metabolized in animals and has accumulated residual toxicity. There have been many poisoning incidents at home and abroad. [0003] When a person consumes animal products with "clenbuterol" residues, a serious poisoning reaction occurs. The symptoms are increased skeletal muscle contraction, destroying the fusion between fast-twitch muscle fibers and slow-twitch muscle fibers, causing muscle tremor, especially in the limbs and muscles. Other sympto...

Claims

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Application Information

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IPC IPC(8): G01N30/56G01N33/53G01N33/558
Inventor 熊勇华张波王迪杨晓慧杨曙明于洪侠
Owner 北京中一茶文化发展有限公司
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