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Nitenpyram hapten, artificial antigen and antibody, their preparation methods and application thereof

A technology of nitenpyram and hapten, which is applied in the field of immunochemistry and residue analysis of pesticide small molecule compounds, and achieves high synthesis efficiency, simple synthesis method and complete retention

Inactive Publication Date: 2014-08-13
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nitenpyram artificial antigens and specific antibodies and the establishment of immunoassay methods based on them have not been reported yet

Method used

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  • Nitenpyram hapten, artificial antigen and antibody, their preparation methods and application thereof
  • Nitenpyram hapten, artificial antigen and antibody, their preparation methods and application thereof
  • Nitenpyram hapten, artificial antigen and antibody, their preparation methods and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0062] (1) Preparation of Nitenpyram hapten

[0063] Dissolve 5.400g of nitenpyram in 40ml of methanol and put it into a three-necked flask; add 2.200g of KOH and stir it with a magnetic force to dissolve it; weigh 2.100g of 3-mercaptopropionic acid, dissolve it in 10ml of methanol and transfer it to a 50ml constant pressure dropping funnel Slowly add it dropwise into a three-necked flask, and slowly raise the temperature of the oil bath to 102°C. After the reaction is complete, remove the oil bath. After the reaction solution is naturally cooled to room temperature, evaporate the solvent to dryness with a rotary evaporator to obtain the target product.

[0064] (2) Preparation of Nitenpyram Immunogen

[0065] First, pipette 1mL of anhydrous dimethylformamide into a 4mL small brown bottle; weigh 5.100mg of nitenpyram hapten and dissolve it in the above-mentioned dimethylformamide solution, and ultrasonically dissolve it for 5 minutes to fully dissolve it. 15.700mg of N,N'dicy...

Embodiment 2

[0071] (1) Preparation of Nitenpyram hapten

[0072] Dissolve 5.500g of nitenpyram in 40ml of methanol and put it into a three-necked flask; add 2.200g of KOH and stir it with a magnetic force to dissolve it; weigh 2.100g of 3-mercaptopropionic acid, dissolve it in 10ml of methanol and transfer it to a 50ml constant pressure dropping funnel Slowly add it dropwise into a three-necked flask, and slowly raise the temperature of the oil bath to 100°C. After the reaction is complete, remove the oil bath. After the reaction solution is naturally cooled to room temperature, evaporate the solvent to dryness with a rotary evaporator to obtain the target product.

[0073] (2) Preparation of Nitenpyram Immunogen

[0074] First, pipette 1mL of anhydrous dimethylformamide into a 4mL small brown bottle; weigh 5.200mg of nitenpyram hapten and dissolve it in the above-mentioned dimethylformamide solution, and ultrasonically dissolve it for 5 minutes to fully dissolve it. 15.660mg of N,N'dicy...

Embodiment 3

[0080] (1) Preparation of Nitenpyram hapten

[0081] Dissolve 5.470g of nitenpyram in 40ml of methanol and put it into a three-necked flask; add 2.259g of KOH and stir it with a magnetic force to dissolve it; weigh 2.150g of 3-mercaptopropionic acid, dissolve it in 10ml of methanol and transfer it to a 50ml constant pressure dropping funnel Slowly add it dropwise into a three-necked flask, and slowly raise the temperature of the oil bath to 105°C. After the reaction is complete, remove the oil bath. After the reaction solution is naturally cooled to room temperature, evaporate the solvent to dryness with a rotary evaporator to obtain the target product.

[0082] (2) Preparation of Nitenpyram Immunogen

[0083] First, pipette 1mL of anhydrous dimethylformamide into a 4mL small brown bottle; weigh 5.180mg of nitenpyram hapten and dissolve it in the above-mentioned dimethylformamide solution, and ultrasonically dissolve it fully for 5 minutes. 15.600mg N,N'dicyclohexylcarbodiimi...

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Abstract

The invention discloses a pesticide nitenpyram hapten, artificial antigen and antibody, their preparation methods and application thereof, relating to preparation of artificial hapten, immunogen, coating antigen and antibody of nicotine pesticide nitenpyram small molecule compound having 6-carboxyl ethyl mercapto molecular structure and establishment of immunoassay method. According to the invention, the problems of tedious and complex property, high cost, and slow analysis speed in traditional physical and chemical analysis method, and the invention provides a simple, rapid, sensitive and accurate immunoassay technology, characterized by taking (E)-N-(6-carboxyl ethyl mercapto)-3-pyridine methyl)-N-ethyl-N' -methyl-2-nitro ethylidene diamine as the hapten, and respectively connecting with bovine serum albumin and ovalbumin to synthesize the immunogen and the coating antigen; and carrying out animal immunization on the artificial antigen, drawing blood, separating antiserum out, and purifying to obtain the antibody. The nitenpyram immunodetection method established by using the antibody has good specificity and sensitivity, the detection limit can stably reach 0.20+ / -0.02ng L <-1>, and the synthetic method of the hapten is simple and has good application prospect.

Description

technical field [0001] The invention belongs to the technical field of immunochemistry and residue analysis of small-molecule pesticide compounds (molecular weight less than 1000 Daltons); relates to organic synthesis, immunochemistry, biochemistry and physicochemical testing techniques, etc.; particularly relates to pesticides with 6-carboxyethylmercapto molecular structure Design and synthesis of nitenpyram small-molecule compound hapten and artificial antigen, preparation of specific antibody for immunized animals and establishment of immunoassay method. Background technique [0002] Nitenpyram, also known as imidachlor, is the second-generation neonicotinoid insecticide after imidacloprid and one of the latest neonicotinoid insecticides. Its insecticidal mechanism is similar to that of traditional insecticides. different. Nitenpyram acts on the posterior membrane of the insect's nerve junction, and by combining with the acetylcholine receptor, the insect is extremely ex...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D213/70C07K14/765C07K14/77C07K1/107C07K16/44G01N33/543
Inventor 王俊平生威张昕哲王硕张燕
Owner TIANJIN UNIV OF SCI & TECH
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