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Flutriafol hapten, flutriafol antigen, flutriafol antibody, flutriafol detection device and preparation and application thereof

A technology of triadol and hapten, which is applied in the field of immunological detection of food safety, can solve the problems of high residue, complex process and tediousness, etc., and achieve the effect of simple and effective synthesis steps, simple operation process and high reaction yield

Active Publication Date: 2022-07-01
SHENZHEN BIOEASY BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although triadol is a low-toxicity insecticide, improper use will also cause excessive residues in crops, which will cause harm to human health
Studies have confirmed that triadol can cause many symptoms such as convulsions, tremors and uncoordinated movements when taken orally in a single high dose
[0003] Residue analysis and safety evaluation of triadol on crops such as rice, cabbage, cowpea and cotton have been reported at home and abroad. Most of the pretreatments are extracted with reagents such as acetone, methanol, and petroleum ether. Liquid-liquid extraction or solid-phase extraction column Purification, relatively speaking, the pretreatment is cumbersome, and it is mainly detected by gas chromatography (ECD detector) or liquid chromatography (PDA detector), but the process is complicated and the cost is high, and it is difficult to achieve the purpose of rapid detection. It is not suitable for on-site rapid detection, and cannot meet the needs of on-site supervision and law enforcement by regulatory authorities and testing institutions

Method used

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  • Flutriafol hapten, flutriafol antigen, flutriafol antibody, flutriafol detection device and preparation and application thereof
  • Flutriafol hapten, flutriafol antigen, flutriafol antibody, flutriafol detection device and preparation and application thereof
  • Flutriafol hapten, flutriafol antigen, flutriafol antibody, flutriafol detection device and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] In this example, dimethyl sulfide is selected from 99% dimethyl sulfide supplied by Sinopharm Chemical Reagent Co., Ltd., concentrated sulfuric acid is selected from 95%~98% sulfuric acid supplied by Sinopharm Chemical Reagent Co., Ltd., and methanol is selected from Sinopharm Chemical Co., Ltd. Analytical pure methanol supplied by Reagent Co., Ltd., tert-butanol selected from Sinopharm Chemical Reagent Co., Ltd., analytically pure tert-butanol, potassium hydroxide, analytically pure potassium hydroxide supplied by Sinopharm Chemical Reagent Co., Ltd., and anhydrous sodium sulfate selected 99% anhydrous sodium sulfate supplied by Sinopharm Chemical Reagent Co., Ltd., 60% sodium hydride supplied by Sinopharm Chemical Reagent Co., Ltd., 60% sodium hydride supplied by Sinopharm Chemical Reagent Co., Ltd., and analytical pure sodium hydroxide supplied by Sinopharm Chemical Reagent Co., Ltd. Sodium hydroxide, the following examples 2-10 all use the reagents in this example 1....

Embodiment 2

[0035] Example 2 Synthesis and identification of fenconazole hapten

[0036] The preparation method of fenconazole hapten, comprises the following steps:

[0037] S1. Add dimethyl sulfide to the reaction vessel, and slowly add concentrated sulfuric acid and methanol dropwise at 5°C. After 4.5 hours of reaction at room temperature, a first reaction solution is obtained, in which the mixture of dimethyl sulfide, concentrated sulfuric acid and methanol is obtained. The molar ratio is 1:(0.5-1.5):(0.2-1);

[0038] S2. in the first reaction solution, add 2,4-difluorobenzophenone, then add tert-butanol to help dissolve, add potassium hydroxide particles in batches, and the mol ratio of 2,4-difluorobenzophenone and potassium hydroxide is 1: (5-10), react overnight at 45°C to obtain the second reaction solution, pour the second reaction solution into purified water, add ethyl acetate for extraction 3 times, wash with saturated brine 2 times, anhydrous After drying with sodium sulfat...

Embodiment 3

[0044] Example 3 Preparation and identification of antigens for immunization and coating of fenconazole

[0045] 3.1 Preparation of Antigen for Fenconazole immunization: Weigh 30mg of Fenconazole hapten prepared in Example 2, dissolve it in 2ml of N,N-dimethylformamide (DMF), add 18mg of N-hydroxybutanediol imide (NHS), 26 mg 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC.HCl), react at room temperature for 6 h to prepare an activation solution; 60 mg bovine serum albumin ( BSA) was dissolved in 2ml of 0.05M boric acid buffer at pH 9.0, added 1ml of DMF, 0.5ml of the above activation solution, reacted at room temperature for 4h, and then dialyzed with PBS (0.02 mol / L, pH=7.4 phosphate buffer), every 4 hours h Change the medium once, change the medium 7-8 times, centrifuge at 4000 rpm for 5 min after dialysis, and take the supernatant to obtain the falconazole hapten-BSA conjugate, which is the antigen for falconol immunization, which is then packaged in aliquo...

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Abstract

The invention discloses a flutriafol hapten, a flutriafol antigen, a flutriafol antibody, a flutriafol detection device as well as preparation and application of the flutriafol hapten, the flutriafol antigen, the flutriafol antibody and the flutriafol colloidal gold chromatography detection device as well as preparation and application of the flutriafol colloidal gold chromatography detection device in flutriafol detection. An immunological detection method is applied to exclusively detect flutriafol remaining in a sample, and the method has the advantages of being good in specificity, high in sensitivity, easy and convenient to operate, capable of achieving on-site rapid detection and the like, and can better meet the on-site supervision and law enforcement requirements of supervision departments and detection organizations; and a technical support is provided for related law-enforcing departments to rapidly detect whether flutriafol is left in vegetables and fruits.

Description

technical field [0001] The present invention relates to the technical field of immunological detection of food safety, and more particularly, the present invention relates to a triconazole hapten, an antigen, an antibody, a detection device and preparation and application thereof. The detection device is especially suitable for triconazole in vegetables and fruits. Residual rapid detection. Background technique [0002] As a low-toxicity and high-efficiency insecticide, fenconazole is widely used and can effectively control various pests on crops such as grain, cotton, fruit and vegetables. Although triconazole is a low-toxicity insecticide, improper use can also cause excessive residues in crops, thereby causing harm to human health. Studies have confirmed that a single high-dose oral dose of fenconazole can cause many symptoms such as spasms, tremors, and incoordination in the human body. [0003] Residue analysis and safety evaluation research of fentriazole on rice, ca...

Claims

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Application Information

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IPC IPC(8): C07D249/10C07K14/765C07K14/77C07K16/44G01N33/577G01N33/558G01N33/58G01N33/53
CPCC07D249/10C07K14/765C07K14/77C07K16/44G01N33/577G01N33/558G01N33/587G01N33/5308G01N2430/10Y02P20/55
Inventor 王炳志杨星星付辉严义勇邓启福杨中程令张鑫王水树肖昭理
Owner SHENZHEN BIOEASY BIOTECHNOLOGY CO LTD
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