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Preparation method and application of hapten and artificial antigen capable of being used for detecting crystal violet and malachite green together

A malachite green and artificial antigen technology, applied in the field of immunochemistry, can solve the problems of potential safety hazards of raw materials, poor stability of hapten, large difference between batches of antigens, etc., and the temperature requirements are not harsh, the product properties are stable, and the cost low effect

Inactive Publication Date: 2016-04-20
广州润坤生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The object of the present invention is to provide a kind of hapten that can be used in common detection crystal violet and malachite green, the preparation method of artificial antigen and its application, to solve the potential safety hazard of the raw material prepared in the prior art, poor stability of hapten, The problem of large difference between antigen batches and high cost, while ensuring high cross-reactivity to malachite green and crystal violet

Method used

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  • Preparation method and application of hapten and artificial antigen capable of being used for detecting crystal violet and malachite green together
  • Preparation method and application of hapten and artificial antigen capable of being used for detecting crystal violet and malachite green together
  • Preparation method and application of hapten and artificial antigen capable of being used for detecting crystal violet and malachite green together

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] (1) Get 2.0g (14.8mmol) of 4-methylaminobenzaldehyde in a 50ml round bottom flask, then add 20ml dehydrated alcohol successively, 3.7g (22.2mmol) of ethyl 2-bromoacetate and 2.0g of potassium carbonate ( 14.8 mmol). Reflux at 80-90°C for 3h. After the reaction is complete, remove the solvent under reduced pressure, add 40ml of purified water, extract twice with ethyl acetate, remove the solvent under reduced pressure, crystallize petroleum ether to obtain the desired compound a 1 2.6g.

[0057] (2) 2.6g compound a 1 Dissolve in 13ml of ethanol, add 3.6g (29.4mmol) of N,N-dimethylaniline and 4.0g (29.4mmol) of zinc chloride, and reflux at 80-90°C for 24h. After the reaction was completed, the solvent was removed under reduced pressure. Add 40ml of purified water, extract with ethyl acetate, remove the solvent under reduced pressure, and purify by column chromatography to obtain the desired compound b 1 2.3 g (ethyl acetate / petroleum ether, 1 / 10, v / v).

[0058] (3) ...

Embodiment 2

[0062] (1) Take 2.0g (14.8mmol) of 4-methylaminobenzaldehyde in a 50ml round bottom flask, then add 20ml of absolute ethanol, 4.3g (22.2mmol) of ethyl 4-bromobutyrate and 2.0g of potassium carbonate (14.8 mmol). Reflux at 80-90°C for 3h. After the reaction is complete, remove the solvent under reduced pressure, add 40ml of purified water, extract twice with ethyl acetate, remove the solvent under reduced pressure, crystallize petroleum ether to obtain the desired compound a 2 2.9g.

[0063] (2) 2.9g compound a 2 Dissolve in 13ml of ethanol, add 3.5g (29.1mmol) of N,N-dimethylaniline and 4.0g (29.1mmol) of zinc chloride, and reflux at 80-90°C for 24h. After the reaction was completed, the solvent was removed under reduced pressure. Add 40ml of purified water, extract with ethyl acetate, remove the solvent under reduced pressure, and purify by column chromatography to obtain the desired compound b 2 2.0 g (ethyl acetate / petroleum ether, 1 / 10, v / v).

[0064] (3) 2.0g compou...

Embodiment 3

[0068] (1) Take 2.0g (14.8mmol) of 4-methylaminobenzaldehyde in a 50ml round bottom flask, then add 20ml of absolute ethanol, 5.0g (22.2mmol) of ethyl 6-bromohexanoate and 2.0g of potassium carbonate (14.8 mmol). Reflux at 80-90°C for 3h. After the reaction is complete, remove the solvent under reduced pressure, add 40ml of purified water, extract twice with ethyl acetate, remove the solvent under reduced pressure, crystallize petroleum ether to obtain the desired compound a 3 2.7g.

[0069] (2) 2.7g compound a 3 Dissolve in 13ml of ethanol, add 2.9g (24.3mmol) of N,N-dimethylaniline and 3.3g (24.3mmol) of zinc chloride, and reflux at 80-90°C for 24h. After the reaction was completed, the solvent was removed under reduced pressure. Add 40ml of purified water, extract with ethyl acetate, remove the solvent under reduced pressure, and purify by column chromatography to obtain the desired compound b 3 2.6 g (ethyl acetate / petroleum ether, 1 / 10, v / v).

[0070] (3) 2.6g compo...

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Abstract

The invention discloses a preparation method and application of hapten and artificial antigen capable of being used for detecting crystal violet and malachite green together. The hapten is a compound shown in the formula (IV), and the antigen is formed by coupling the hapten with carrier protein and used for crystal violet and malachite green. The antigen and the antibody prepared through the method can be used for preparing enzyme-linked immunoassay kit. The method has the advantages that the production process is safe and simple, temperature requirements and conditions are not strict, a product is stable in quality, and cost is low; according to the hapten, the antigen and the corresponding antibody structure of the antigen, the prepared enzyme-linked immunoassay kit is high in sensitivity to malachite green and crystal violet, and the cross reaction rate reaches 95% or more.

Description

technical field [0001] The invention relates to the technical field of immunochemistry, in particular to a method for preparing a hapten and an antigen that can be jointly used to detect crystal violet and malachite green. Background technique [0002] Both crystal violet and malachite green are toxic triphenylmethane chemicals that are fungicides, parasites and carcinogens. At present, the main method for residual detection of crystal violet and malachite green is to adopt gas chromatography (GC), gas chromatography mass spectrometry (GCMS), liquid chromatography (HPLC) or liquid phase mass spectrometry (LCMS). Although the above methods can be accurately quantified, due to equipment The instrument is expensive, the detection time is long, and professional personnel are required to operate, so it is impossible to realize the real on-site detection. Immunoassay detection technology is a new rapid and accurate detection method developed in the fields of environment and food ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C229/18C07C227/18C07K14/795C07K14/77C07K14/765G01N33/577
CPCC07C227/04C07C227/16C07C227/18C07C229/18C07K14/765C07K14/77C07K14/795G01N33/577
Inventor 胡睿李斌
Owner 广州润坤生物科技有限公司
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