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Synthesizing porcess for artificial antigen of cyanobromide chrysanthemum ester and assaying process thereof

A technology of artificial antigen and synthesis method, which is applied in the direction of biological testing, material inspection products, etc., can solve the problems that there is no rapid detection method yet, and achieve the effects of convenient and fast processing methods, reducing pollution, and reducing agricultural costs

Inactive Publication Date: 2012-07-04
BEIJING UNIV OF AGRI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Deltamethrin is the most widely used pyrethroid pesticide in China. The immunoassay of deltamethrin in domestic research is in the preliminary stage, and there is no rapid detection method for it yet.

Method used

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  • Synthesizing porcess for artificial antigen of cyanobromide chrysanthemum ester and assaying process thereof
  • Synthesizing porcess for artificial antigen of cyanobromide chrysanthemum ester and assaying process thereof
  • Synthesizing porcess for artificial antigen of cyanobromide chrysanthemum ester and assaying process thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Total synthesis of deltamethrin artificial hapten

[0029] Carry out the total synthesis of deltamethrin artificial hapten according to the following chemical reaction formula:

[0030] 1. Total synthesis of deltamethrin artificial hapten

[0031]

[0032] In the formula:

[0033] 1: Trihydroxybenzaldehyde

[0034] 2: p-Nitrochlorobenzene

[0035] 3: 3-(4-nitrophenoxy)benzaldehyde

[0036]4: [3-(4-nitrophenoxy)phenyl]cyanomethanol

[0037] 5: (1R,3R)-3-(2,2-dibromovinyl)-2,2-dimethylcyclopropylcarboxylate [3-(4-nitrophenoxy)phenyl]cyanomethyl ester

[0038] 6: (1R,3R)-3-(2,2-dibromovinyl)-2,2-dimethylcyclopropylcarboxylate [3-(4-aminophenoxy)phenyl]cyanomethyl ester . 6 is an artificial hapten, which is linked to the protein to obtain the antigen.

[0039] The specific steps are:

[0040] 1), the synthesis of (1R, 3R)-3-(2,2-dibromoethenyl)-2,2-dimethylcyclopropylformyl chloride: add 30g (1R, 3R)- 30ml of chloroform solution of 3-(2,2-dibromovi...

Embodiment 2

[0045] Example 2 Synthesis of deltamethrin artificial antigen

[0046] Carry out according to the following chemical reaction formula:

[0047]

[0048] The specific steps are:

[0049] Weigh 0.0261g of the hapten (DM, 0.05mmol) into a 50ml small beaker, add four drops of absolute ethanol in an ice bath, and stir to dissolve. Then add 0.6ml of 1M hydrochloric acid and 0.4ml of 0.2M sodium nitrite to the solution, and stir well. Finally, 0.4 ml of DMF was added dropwise to the above homogeneous reaction solution.

[0050] Weigh 45mg of bovine serum albumin (BSA) or chicken egg serum albumin (OVA) into a 50ml small beaker, add a mixture of 5ml boric acid (0.2M) buffer solution (ph=8.7) and 1.2ml DMF to dissolve. Under ice-bath conditions, the active hapten solution was added dropwise to the stirring protein, and the dropwise addition was completed within 50 minutes. The reaction mixture was stirred in an ice bath to continue the reaction for 45 minutes, and was put into a...

Embodiment 3

[0051] Example 3 Determination of Antigen

[0052] Weighed 21.7 mg of the synthesized deltamethrin hapten (DM), diluted it with absolute ethanol, prepared into series concentrations of 0.0283, 0.0567, 0.1131, 0.2263, and 0.4525 mg / ml, and carried out UV spectrum scanning respectively; Read the absorbance Amax at the maximum absorption peak of the hapten on the spectrogram, and use it as the DM concentration (C DM ) versus absorbance standard absorbance curve.

[0053] Weigh 1.28 mg of BSA, dilute it in phosphate buffer solution (PH=7.4), and make it into series concentrations of 0.015, 0.027, 0.053, 0.107, 0.214 mg / ml, and scan the UV spectrum respectively; read out from the UV spectrum of BSA Absorbance Amax at the maximum absorption peak of bovine serum albumin was used as BSA concentration (C BSA ) versus absorbance standard absorbance curve.

[0054] The dialyzed hapten and bovine serum albumin conjugate (DM-BSA) were tested by the Lowry method [10] The protein concent...

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PUM

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Abstract

The invention discloses Dehamethrin artificial antigen synthetic method. It includes the synthesis for (1R, 3R)-3-(2, 2-dibrom vinyl)-2, 2-dimethyl cyclopropyl formyl chloride, 3-(4-nitro phenoxy) benzaldehyde, [3-(4- nitro phenoxy) phenyl] cyan methanol, (1R, 3R)-3-(2, 2-dibrom vinyl)-2, 2-dimethyl cyclopropyl formic acid[3-(4-nitro phenoxy)phenyl] cyan methyl ester, artificial half antigen (1R,3R)-3-(2, 2-dibrom vinyl)-2, 2-dimethyl cyclopropyl formic acid[3-(4-aminophenoxy) phenyl] cyan methyl ester, artificial antigen synthetic steps and its measuring.

Description

technical field [0001] The invention relates to a method for synthesizing and measuring an artificial antigen of deltamethrin. Background technique [0002] Pyrethroids are an important class of insecticides, which are widely used in the control of pests in tea gardens, orchards, and farmland due to their high efficiency, low toxicity, fast knockdown, and less residue. They are currently found to be polluting surface water sources, affecting the environment and causing ecological damage. [0003] Since my country's accession to the WTO, the green barriers have been strict, and the problem of pyrethroid pesticide residues has become the main obstacle hindering the export trade of agricultural and sideline products, especially tea and fruits. Therefore, it is very necessary to establish a sensitive, selective and rapid immunoassay method for detecting pyrethroid pesticide residues. [0004] In order to establish an immunoassay method, it is first necessary to prepare antibod...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53
Inventor 赵建庄魏朝俊王春娜王静
Owner BEIJING UNIV OF AGRI
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