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Production method and use for dichloro quinolinic acid artificial hapten, artificial antigen and specific antibody

A technology of quinclorac and artificial hapten, which is applied in the direction of chemical instruments and methods, specific peptides, material inspection products, etc., can solve problems such as unsuitable for large-scale sample detection and analysis, easy to explode, and human danger, etc., to achieve convenient On-site monitoring, high sensitivity and strong specificity

Inactive Publication Date: 2005-01-26
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Liquid chromatography is mainly for water samples and soil samples with less impurities, and the pretreatment is relatively simple; for those crop samples with more impurities, gas chromatography (ECD detector) or gas-mass spectrometry must be used for detection , the pretreatment is very troublesome, and diazotization treatment must be carried out. The diazomethane and ether solution used are easy to explode, dangerous to people, and not suitable for the detection and analysis of large quantities of samples

Method used

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  • Production method and use for dichloro quinolinic acid artificial hapten, artificial antigen and specific antibody
  • Production method and use for dichloro quinolinic acid artificial hapten, artificial antigen and specific antibody
  • Production method and use for dichloro quinolinic acid artificial hapten, artificial antigen and specific antibody

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preparation example Construction

[0041] 3 Preparation of antibody and establishment of quinclorac ELISA method

[0042]Three kinds of immunogen complexes (Q-BSA, QC-BSA, QB-BSA) were used to immunize three rabbits according to conventional methods. From the second time of booster immunization, blood was collected from the rabbit's ear vein on the 8th day after each immunization, and the titer of the serum was determined by indirect ELISA after appropriate dilution. By the time of the fourth immunization, the rabbits obtained high-titer antibodies, and the titers after purification and freeze-dried powder were 2000, 1.2×10 6 , 1.6×10 6 . Since the titer of anti-Q-BSA antibody was too low, no further experiments will be performed.

[0043] Using quinclorac antigen antibody immune reaction and enzymatic reaction to establish quinclorac ELISA method. It has high specificity and sensitivity in the detection and analysis of quinclorac residues in food, plants, environmental soil and water samples, the lowest de...

Embodiment 1

[0044] Example 1: Synthesis of Hapten QC

[0045] 1) Dilute 37.5ml of SOCl in excess of thionyl chloride 2 And 2.42g (10mmol) quinclorac is packed in the there-necked flask, under magnetic stirring, reflux reaction 1.5h, distillation removes excess SOCl 2 , That is, the product 3,7-dichloro-8-quinoline formyl chloride.

[0046] 2) According to the feeding ratio of 6-aminocaproic acid: 3,7-dichloro-8-quinoline formyl chloride = 1:1, put 1.32g (10mmol) of 6-aminocaproic acid into a three-necked flask, and add 5ml of 2mol / L of NaOH solution, stirred to dissolve, 2.60g (10mmol) 3,7-dichloro-8-quinoline formyl chloride was dissolved in 30ml of dioxane, added to the constant pressure dropping funnel, and the solution was added dropwise five times , and then add 2mol / L NaOH solution to keep the pH value of the reaction solution at 8-10. After the addition is completed, continue to react at 0-4°C for 2h. Rise to room temperature, adjust the pH value of the reaction solution to abo...

Embodiment 2

[0049] Example 2: Synthesis of Hapten QC

[0050] 1) Dilute 37.5ml of SOCl in excess of thionyl chloride 2 And 2.42g (10mmol) quinclorac is packed in the there-necked flask, under magnetic stirring, reflux reaction 1.5h, distillation removes excess SOCl 2 , That is, the product 3,7-dichloro-8-quinoline formyl chloride.

[0051] 2) According to the feeding ratio of 6-aminocaproic acid: 3,7-dichloro-8-quinoline formyl chloride = 1.2: 1, put 1.584g (12mmol) of 6-aminocaproic acid into a three-necked flask, add 6ml2mol / L of NaOH solution, stirred and dissolved, 2.60g (10mmol) 3,7-dichloro-8-quinoline formyl chloride was dissolved in 30ml of dioxane, added to the constant pressure dropping funnel, and added dropwise to the solution five times, Then add 2mol / L NaOH solution to keep the pH value of the reaction solution at 8-10. After the addition is complete, continue to react at 0-4°C for 3h. Rise to room temperature, adjust the pH value of the reaction solution to about 4.0 wi...

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Abstract

The invention discloses a process for preparing Quinclorac artificial semiantigen, artificial antigen, specific antibody and use thereof, wherein the preparation comprises, subjecting the dichloroquine (3,7-dichlorine-8-quinoline carboxylic acid) to sulfoxide chlorinated acylation, reacting with reanal and aminocaproic acid, thus obtaining semiantigen 4-(3,7-dichlorine-8-quinolineformyl) reanal or 6-(3,7-dichlorine-8-quinolineformyl) aminocaproic acid (QB or QC).

Description

technical field [0001] The invention relates to a quinclorac artificial hapten, artificial antigen, specific antibody preparation method and application thereof, belonging to the technical field of pesticide immunochemistry. Background technique [0002] Due to the continuous expansion of the scale of pesticide use, the environmental impact of pesticide residues and the chronic and long-term effects on human health are increasingly concerned and worried, and the restrictions on pesticide residues are therefore becoming more and more stringent. New requirements and higher standards have been put forward in various aspects such as indicators and indicators. For small molecular toxic chemicals with a molecular weight less than 1000 Daltons, such as pesticides and their metabolites, traditional residue analysis methods mainly rely on physical and chemical analysis methods such as gas chromatography (GC), liquid chromatography (HPLC) or mass spectrometry (MS). Traditional physic...

Claims

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Application Information

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IPC IPC(8): C07D215/18C07K16/00G01N33/02G01N33/563
Inventor 朱国念程敬丽魏方林施海燕吴慧明
Owner ZHEJIANG UNIV
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