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Polyclone antibody of H-9201, and preparation method and application thereof

A technology of H-9201 and metolaphos, which is applied in chemical instruments and methods, compounds of Group 5/15 elements of the periodic table, and material testing products, etc., to achieve the effect of a simple preparation method

Inactive Publication Date: 2012-07-18
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, research reports on the detection of bimethafos (H-9201) residues have only been found in gas chromatography (see Zhang Cunzheng, Zhang Zhiyong, Liu Yuan, Wang Donglan, Liu Xianjin. The new herbicide bimethafos (H-9201) has Research on Residue Dynamics in Soil. Journal of Analytical Science, 2007, 23(3): 337-339.), the preparation of antibody and immunoassay for Dimethafos (H-9201) and its application in residue detection , there is no literature report at home and abroad

Method used

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  • Polyclone antibody of H-9201, and preparation method and application thereof
  • Polyclone antibody of H-9201, and preparation method and application thereof
  • Polyclone antibody of H-9201, and preparation method and application thereof

Examples

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Embodiment 1

[0037] Example 1. Preparation of Amisofos Transition State Similar Compounds

[0038] Phosphor trichloride and methanol are used as raw materials, the temperature of the solution is controlled below -5°C for reaction, the solution is washed with ice water and the lower layer is retained to obtain methoxyphosphoryl thiodichloride.

[0039] Under the temperature-controlled condition of acetone solution at 5-10°C, triethylamine is used as a catalyst, methoxyphosphoryl thiol) and 2,4-dimethyl-6-nitrophenol (DMNT), sodium hydroxide (NaOH ) reaction to obtain an acetone solution containing the compound of molecular structural formula (5).

[0040] The compound solution containing the molecular structural formula (5) is pre-cooled to 5-10°C, and ammonia water is added dropwise thereto. After the ammonia water is added dropwise, the pH is about 8, add 40% aqueous sodium hydroxide solution dropwise until the pH is 10, remove the acetone, add 30 mL of diethyl ether to extract twice, wa...

Embodiment 2

[0042] Embodiment 2. Preparation of the compound of molecular structural formula (3)

[0043] Mix 10ml of 0.5mM dimethazone transition state analogue compound with 0.8mM succinic anhydride acetonitrile solution, add dropwise 0.5mM DMAP acetonitrile solution, and stir for 2 hours at 50°C; after the reaction is terminated, add 50ml of distilled water, Extraction with methyl chloride, dehydration of the organic phase with anhydrous magnesium sulfate, and removal of the organic solvent to obtain the compound of molecular structure formula (3). Its liquid chromatography / mass spectrometry identification chart (LC / MS) see Figure 4 .

Embodiment 3

[0044] Example 3. Synthesis of antigens

[0045] Add 0.23mM NHS, 0.33mM DCC, 0.033mM DMAP to the dichloromethane solution containing 0.3mM compound of molecular formula (3), stir and react overnight at 0°C; centrifuge at 10000g for 10min the next day, discard the precipitate, and remove Solvent, dissolve the obtained activated ester with DMF, slowly add the activated ester solution to 5 mL of pre-cooled phosphate buffer containing 40 mg bovine serum albumin BSA, the pH of the phosphate buffer is 9.0, and react overnight at 4°C;

[0046] After the reaction, put the reaction mixture into a dialysis bag, dialyze in deionized water at 4°C for 3 days, change the water every 8 hours, obtain the immune antigen with molecular structure formula (1) after the dialysis, subpackage and store in - Store in the refrigerator at 20°C for later use. Such as Figure 5 Figure 6 The UV scanning spectrum shown shows that the maximum absorption peaks of hapten (hapten) and carrier protein (BSA ...

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Abstract

The invention relates to a polyclone antibody of H-9201, and a preparation method and application thereof. The preparation method comprises the following steps: coupling hapten and bovine serum albumin (BSA) to obtain an immune antigen, and immunizing rabbits in New Zealand to obtain the polyclone antibody. The invention is characterized in that the hapten is prepared from introducing carboxyl group into H-9201 transition state analog compound which has a molecular structural formula (4); the hapten and carrier protein are coupled by using an active ester method to obtain a complete antigen; the polyclone antibody is obtained by animal immunization; and an ELISA detection method is established in the invention. The invention can be used for detecting residues of pesticide H-9201 in the water body sample. The polyclone antibody of H-9201 can be used for quickly detecting residues of massive pesticide H-9201 sample in a water body.

Description

Technical field: [0001] The invention relates to a preparation method and application of bimethafos (H-9201) polyclonal antibody. Background technique: [0002] Dimethazone (H-9201) is a new type of agricultural herbicide independently developed by my country with independent intellectual property rights. Compound, its chemical name is: O-methyl-O-(2,4-dimethyl-6-nitrophenoxy)-N-isopropylthiophosphoramidate, molecular weight is 318.5, is a kind of Phosphoramidosulfurized ester compounds, the trade name is Dimethofos. The compound is a systemic conduction type selective soil treatment herbicide for both water and drought, which can be used for soybean, rice, wheat, corn, vegetables and other crops to prevent and control a variety of annual monocotyledonous and dicotyledonous weeds, and has a wide herbicidal spectrum. And it can be compounded with a variety of herbicide varieties. Field efficacy tests have not found any impact on the growth of subsequent crops. It is a new her...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/44C07K16/06G01N33/531G01N33/53C07F9/24
Inventor 张存政刘贤进杨春龙陈敏俞杰
Owner JIANGSU ACAD OF AGRI SCI
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