Imidacloprid hapten, complete antigen and its preparation method and application

A complete antigen and imidacloprid technology, applied in the biological field, can solve problems such as long pesticide residues, human and livestock threats, agricultural products, food sales and safety export impact, and achieve high sensitivity, simple preparation method, and strong specificity Effect

Active Publication Date: 2018-03-16
FUJIAN ANXIN RUIJIE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the effect of imidacloprid is remarkable and the pests can be effectively controlled after use, the long residue of the pesticide will pose a threat to humans and livestock, and affect the sales and safe export of agricultural products and food

Method used

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  • Imidacloprid hapten, complete antigen and its preparation method and application
  • Imidacloprid hapten, complete antigen and its preparation method and application
  • Imidacloprid hapten, complete antigen and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A preparation method of imidacloprid hapten, comprising the following steps:

[0029] 1) Dissolve 1 part of 6-methylnicotinic acid in 4 parts of methanol, add 0.2 parts of concentrated sulfuric acid, heat the reaction system to reflux at 65°C for 3 hours, and distill off the methanol under reduced pressure to obtain a white solid powder, which is 6-methanol methyl nicotinate;

[0030] 2) Dissolve 1 part of the above-mentioned 6-methyl nicotinic acid methyl ester in 4 parts of carbon tetrachloride, then add 1.5 parts of N-bromosuccinimide, wait for the temperature of the reaction system to rise to 80 ° C, add 0.2 parts of azo Diisobutyronitrile, the reaction system was reacted at 80°C for 3 hours, carbon tetrachloride was distilled off under reduced pressure to obtain a brown liquid, and the brown liquid was purified by column chromatography to obtain a white solid, which was 6-(bromomethyl)nicotinic acid methyl ester;

[0031] 3) Dissolve 1 part of the above 6-(bromom...

Embodiment 2

[0035] A preparation method of imidacloprid hapten, comprising the following steps:

[0036] 1) Dissolve 1 part of 6-methylnicotinic acid in 3 parts of methanol, add 0.1 part of concentrated sulfuric acid, heat the reaction system to reflux at 70°C for 3 hours, and distill off the methanol under reduced pressure to obtain a white solid powder, which is 6- Methyl nicotinate;

[0037] 2) Dissolve 1 part of the above-mentioned 6-methyl nicotinic acid methyl ester in 3 parts of carbon tetrachloride, then add 1.2 parts of N-bromosuccinimide, wait for the temperature of the reaction system to rise to 80 °C, add 0.1 parts of peroxy Benzoyl, the reaction system was reacted at 70°C for 4 hours, carbon tetrachloride was distilled off under reduced pressure to obtain a brown liquid, and the brown liquid was purified by column chromatography to obtain a white solid, which was methyl 6-(bromomethyl)nicotinate ;

[0038] 3) Dissolve 1 part of the above 6-(bromomethyl) nicotinic acid methy...

Embodiment 3

[0041] Prepare imidacloprid complete antigen with the imidacloprid hapten of embodiment 1, comprise the following steps:

[0042] 1) Dissolve 20 μmol imidacloprid hapten in 1 mL DMF, add 60 μmol DCC and 60 μmol NHS, stir at room temperature for 24 hours to obtain liquid A;

[0043] 2) Dissolve 0.4 μmol BSA in 3 mL carbonate buffer to obtain solution B;

[0044] 3) Add the above liquid A dropwise to liquid B and stir at 4°C for 12 hours to obtain liquid C;

[0045] 4) Put liquid C in a dialysis bag and dialyze it in phosphate buffer, change the liquid once every 3 hours, dialyze 6 times in total, collect the solution in the dialysis bag, which is the immunogen solution, that is, the imidacloprid complete antigen,- Store at 20°C.

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Abstract

The invention discloses an imidacloprid hapten, complete antigen and its preparation method and an application. The imidacloprid hapten is prepared by taking 6-methyl nicotinic acid, and reacting withmethanol to generate methyl 6-methylnicotinate; reacting methyl 6-methylnicotinate with N-bromosuccinimide to obtain 6-( bromomethyl) methyl nicotinate, and then reacting with N-( imidazolidine-2- subunit) nitramide to generate 6-((2-(nitroamino)imidazoline-1-radical)methyl) methyl nicotinate; finally, reacting ester to be 6-((2-(nitroamino)imidazoline-1-radical)methyl) nicotinic acid by NaOH solution. The imidacloprid hapten is coupled with carrier protein, and the imidacloprid complete antigen is prepared. The immune animal experiment indicates that the artificial antigen has good immunogenicity; the imidacloprid hapten and imidacloprid antigen can be applied to imidacloprid immunoassay analysis; the application prospect is very extensive.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an imidacloprid hapten, a complete antigen, a preparation method and an application. technical background [0002] Imidacloprid is an ultra-efficient systemic insecticide containing a pyridine ring. It is effective against Thysanoptera, Coleoptera, Homoptera, Lepidoptera and Diptera, especially for piercing-sucking mouthpart pests such as Chinese pear tree Lice, green peach aphid, small green leafhopper, apple tuberculosis, thrips, black-tailed leafhopper, pear aphid, whitefly, pear weevil, etc. have high residual effect and systemic properties, and can be widely used in wheat, vegetables, rice, Most crops such as cotton have the characteristics of high efficiency, low toxicity, and strong broad spectrum. Although imidacloprid has a remarkable effect and can effectively control insect pests after use, the long residue of the pesticide poses a threat to humans and livesto...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D401/06C07K14/765C07K1/107G01N33/535
CPCC07K14/765C07K19/00G01N33/535G01N2430/10
Inventor 王保民陈俊玉王冕谭桂玉刘伟华
Owner FUJIAN ANXIN RUIJIE BIOTECH CO LTD
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