Fluazinam hapten, artificial antigen and antibody and preparation method and application thereof

An artificial antigen, fluazinamide technology, applied in the preparation method of peptide, chemical instrument and method, animal/human protein, etc., to achieve the effect of enhanced immunogenicity, simple reaction operation and strong specificity

Active Publication Date: 2022-07-08
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant report on the fluazinam hapten in China

Method used

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  • Fluazinam hapten, artificial antigen and antibody and preparation method and application thereof
  • Fluazinam hapten, artificial antigen and antibody and preparation method and application thereof
  • Fluazinam hapten, artificial antigen and antibody and preparation method and application thereof

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preparation example Construction

[0029] In a second aspect, the present invention provides a method for preparing the above-mentioned fluazinam hapten, comprising the following steps:

[0030] 1) 2,4-dichloro-3,5-dinitrotrifluorotoluene reacts with methyl 2-aminobutyrate-3-chloro-5-trifluoromethylpyridine to obtain intermediate 1, the intermediate Body 1 has the structural formula

[0031] 2) The intermediate 1 is subjected to a hydrolysis reaction to obtain the fluazinam hapten.

[0032] Preferably, the step 1) includes the following steps: taking 2,4-dichloro-3,5-dinitrotrifluorotoluene and adding an organic solvent to dissolve, adding anhydrous potassium carbonate and 2-aminobutyric acid methyl ester-3 -Chloro-5-trifluoromethylpyridine, heated in an oil bath for reaction, after the reaction was completed, rotary evaporated, added water, extracted with ethyl acetate, collected the organic phase, evaporated the organic phase to dryness, and then used dichloromethane-cyclohexane beating with alkane, sucti...

Embodiment 1

[0051] A kind of preparation method of fluazinam hapten, it comprises the steps:

[0052] 1) Dissolve 0.304g of 2,4-dichloro-3,5-dinitrotrifluorotoluene in 80mL of acetonitrile, add 0.32g of anhydrous potassium carbonate and 0.21g of methyl 2-aminobutyrate-3-chloro-5 -Trifluoromethylpyridine, heated in an oil bath for 12h, after the reaction was completed, the acetonitrile was removed by rotary evaporation, 60mL of water was added, extracted with 80mL of ethyl acetate, the organic phase was collected, and the organic phase was evaporated to dryness with 20mL volume ratio of 1 : 3 of dichloromethane-cyclohexane was slurried, filtered with suction, and washed with n-hexane to obtain Intermediate 1;

[0053] 2) Weigh intermediate 1, add alkaline solution, heat to reflux, and carry out hydrolysis reaction; after the reaction, adjust pH to slightly acidic with hydrochloric acid, then extract with ethyl acetate, and the organic phase is evaporated to dryness and purified by column t...

Embodiment 2

[0055] A kind of preparation method of fluazinam hapten, it comprises the steps:

[0056] 1) Dissolve 0.304g of 2,4-dichloro-3,5-dinitrotrifluorotoluene in 80mL of acetonitrile, add 0.28g of anhydrous potassium carbonate and 0.17g of methyl 2-aminobutyrate-3-chloro-5 -Trifluoromethylpyridine, heated in an oil bath for 12h, after the reaction was completed, the acetonitrile was removed by rotary evaporation, 60mL of water was added, extracted with 80mL of ethyl acetate, the organic phase was collected, and the organic phase was evaporated to dryness with 20mL volume ratio of 1 : 3 of dichloromethane-cyclohexane was slurried, filtered with suction, and washed with n-hexane to obtain Intermediate 1;

[0057] 2) Weigh intermediate 1, add alkaline solution, heat to reflux, and carry out hydrolysis reaction; after the reaction, adjust pH to slightly acidic with hydrochloric acid, then extract with ethyl acetate, and the organic phase is evaporated to dryness and purified by column t...

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Abstract

The invention discloses fluazinam hapten, artificial antigen and antibody, and preparation methods and applications thereof. The fluazinam hapten provided by the invention not only retains the characteristic structure of fluazinam, but also makes the immune The originality is obviously enhanced, and it has a carboxyl group that can be coupled with the carrier protein; the fluazinam immunization antigen obtained by coupling the fluazinam hapten with the carrier protein is used to immunize animals, which is more conducive to stimulating the animal's immune response to produce specificity A stronger and more sensitive antibody, the sensitivity of the antibody to fluazinam can reach 0.1 μg / L, and the cross-reaction rate with other pesticides is low, which provides a basis for the subsequent establishment of various immunoassay methods for fluazinam.

Description

technical field [0001] The invention belongs to the field of food safety detection. More specifically, the present invention relates to fluazinam haptens, artificial antigens and antibodies, and methods for their preparation and use. Background technique [0002] Fluazinam is a 2,6-dinitroaniline low-toxicity and broad-spectrum fungicide independently developed and produced by Ishihara Industry in Japan. It has been used in various crops such as Chinese cabbage, tomato, cucumber, pepper and potato Obtained registration, has good control effect on phytophagous mites and cruciferous plant clubroot. But its environmental toxicity is a concern, and studies have shown that fluazinam can trigger diseases such as asthma and dermatitis that damage the human immune system. [0003] At present, there are mainly analytical methods such as gas chromatography, high performance liquid chromatography and liquid chromatography mass spectrometry for the detection of fluazinam at home and a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D213/74C07K14/765C07K14/77C07K14/795C07K16/44C07K19/00C07K1/02G01N33/53
CPCC07D213/74C07K14/765C07K14/77C07K14/795C07K19/00C07K16/44G01N33/53
Inventor 崔廷婷冯才伟何方洋刘玉梅冯月君魏力杰赵正苗马玉华
Owner BEIJING KWINBON BIOTECH
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