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Isocarbophos semiantigen, and preparation method and application thereof

A kind of hydrophosphine and hapten technology, which is applied in the field of immunity, can solve the problems of no hydrophosphine immunochemical test method, expensive instruments, difficult to popularize and popularize, etc.

Inactive Publication Date: 2016-01-27
SHENZHEN ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods require professional and technical personnel to operate, and the instruments are expensive, not easy to carry, and difficult to promote and popularize.
At present, there are no reports on immunochemical detection methods and products for isocarbophos

Method used

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  • Isocarbophos semiantigen, and preparation method and application thereof
  • Isocarbophos semiantigen, and preparation method and application thereof
  • Isocarbophos semiantigen, and preparation method and application thereof

Examples

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preparation example Construction

[0036] Such as figure 1 The preparation method of shown above-mentioned isocarbophos hapten comprises the steps:

[0037] S110, in the presence of a catalyst and under alkaline conditions, mix and react isopropyl salicylate and O-methyl phosphorothioate dichloride, and obtain O-methyl-O-2-isopropyl salicylate after purification Thiophosphoryl chloride.

[0038] In S110, the specific reaction formula is as follows:

[0039]

[0040] Wherein, the catalyst is a quaternary ammonium salt phase inversion catalyst. The quaternary ammonium salt phase inversion catalyst can be tetrabutylammonium bromide.

[0041] Specifically, the operation of S110 is: dissolve isopropyl salicylate with dichloromethane, add an appropriate amount of quaternary ammonium salt phase transfer catalyst, cool to about 0°C, and add the same amount of O-methyl Thiophosphoryl dichloride, add and rise to room temperature, then add dropwise 20% sodium hydroxide solution of 4 to 6 times the amount of the sub...

Embodiment 1

[0073] The preparation of embodiment 1 isocarbophos hapten

[0074] Synthesis of isocarbophos hapten:

[0075] (1) Put 6g of salicylic acid into a round bottom flask, then add 10mL of concentrated sulfuric acid and 15g of isopropanol in sequence, heat the oil bath to 100°C, condense and reflux for 12h, then cool to room temperature, add water to dilute, extract with ethyl acetate, and combine The organic layer was washed successively with saturated sodium bicarbonate and distilled water, dried with anhydrous sodium sulfate, evaporated to dryness and concentrated to obtain isopropyl salicylate.

[0076] (2) Use a graduated cylinder to measure 10mL of phosphorus trichloride into a round bottom flask, slowly add 3 times the volume of anhydrous methanol at -5°C to 0°C for 3 hours, shake it with ice water for 2 to 3 times and let it stand still. The organic layer was separated and dried with anhydrous sodium sulfate to obtain a colorless and transparent liquid O-methylphosphoryl t...

Embodiment 2

[0080] The preparation of embodiment 2 isocarbophos artificial antigens

[0081] Antigen ICP-H1-BSA: Dissolve 15mg of hapten ICP-H1, 8.8mg of NHS (N-hydroxysuccinimide and 15mg of DCC) in 1mL of anhydrous DMF (N,N-dimethylformamide), and stir the reaction at room temperature After 18 hours, the reaction solution was centrifuged at 4000 rpm for 5 minutes, and the supernatant activation solution was absorbed. Then weigh 120mg BSA and dissolve it in 8mL of carbonate buffer solution (0.05mol / L, pH9.6), slowly add 400μL dropwise for activation solution, then stirred and reacted at room temperature for 3 hours, then put the yellow reaction solution into a dialysis bag, dialyzed with PBS (phosphate buffer saline 0.01mol / L, pH7.4), changed the solution once every 4 hours, and changed the solution 7 to 8 times After dialysis, centrifuge at 4000 rpm for 5 min, and store the supernatant at -20°C.

[0082] Antigen ICP-H2-OVA: Dissolve 18 mg hapten ICP-H2, 8.2 mg NHS (N-hydroxysuccinimide...

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Abstract

A disclosed isocarbophos semiantigen possesses a structural formula shown in the specification. In the formula, -R is -(CH2)5COOH or -PhCOOH. The isocarbophos semiantigen is designed aiming at isocarbophos and is applicable to prepare an isocarbophos antibody. Experiments prove that the prepared isocarbophos antibody is capable of specifically identifying isocarbophos with both relatively good accuracy and sensitivity. The invention also discloses a preparation method and application of the above isocarbophos semiantigen.

Description

technical field [0001] The invention relates to the technical field of immunization, in particular to an isocarbophos hapten and a preparation method and application thereof. Background technique [0002] Isocarbophos (ICP) is a broad-spectrum organophosphorus insecticide and acaricide with high insecticidal efficiency and easy decomposition. It once became one of the main pesticides for controlling pests and diseases in agricultural production; ICP will cause pesticide residues, and through the enrichment of the food chain, it can poison animals and humans, affect the metabolic balance of hormones in the human body, and especially have a more serious impact on the normal development of the nervous system of children. The reported ICP detection mainly includes polarography and chromatography. However, these methods require professional and technical personnel to operate, expensive instruments, not easy to carry, and difficult to promote and popularize. [0003] In recent y...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07F9/24C07K14/765C07K14/77C07K14/795C07K1/107C07K16/44G01N33/531
Inventor 何建安顾大勇杨燕秋杨星星姚旌羽朱海史蕾刘春晓赵纯中徐云庆
Owner SHENZHEN ACAD OF INSPECTION & QUARANTINE
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