Chlorothalonil antigen, antibody preparation method and residual chlorothalonil ELISA (Enzyme-Linked Lmmuno Sorbent Assay) detection method

A technology of chlorothalonil and a new method, applied in the field of pesticide residue enzyme immunoassay, can solve the problems of long detection time and low sensitivity, and achieve the effects of short reaction time, easy control of reaction conditions and high sensitivity

Inactive Publication Date: 2010-09-15
DALIAN NATIONALITIES UNIVERSITY
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  • Chlorothalonil antigen, antibody preparation method and residual chlorothalonil ELISA (Enzyme-Linked Lmmuno Sorbent Assay) detection method
  • Chlorothalonil antigen, antibody preparation method and residual chlorothalonil ELISA (Enzyme-Linked Lmmuno Sorbent Assay) detection method
  • Chlorothalonil antigen, antibody preparation method and residual chlorothalonil ELISA (Enzyme-Linked Lmmuno Sorbent Assay) detection method

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[0039]The present invention will be further clarified through the detailed description of specific embodiments below, but it is not intended to limit the present invention, but only for illustration.

[0040] 1. Molecular design and synthesis of hapten: For the synthesis route of chlorothalonil hapten and artificial complete antigen, see figure 1 . Accurately weigh 100.0mg (0.37mmol) of dried chlorothalonil and dissolve it in 20.0mL of dichloromethane, add ethylenediamine-water mixture (4.0mL-20.0mL) to it, the reaction system forms two phases, stir slowly After 24 hours, the dichloromethane layer was taken, washed with water (10 mL×3), and the solvent was removed by a rotary evaporator to obtain a yellow-green powder. Recrystallization from acetone afforded pure material with figure 2 Infrared spectra shown, such as image 3 The mass spectrum shown in Figure 4 is 1 H-NMR, such as Figure 5 shown 13 C-NMR, so it was identified as 2,4,5-trichloro-6-(2-aminoethylimino)-i...

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Abstract

The invention discloses a chlorothalonil antigen, an antibody preparation method and a residual chlorothalonil ELISA (Enzyme-Linked Lmmuno Sorbent Assay) detection method. Chlorothalonil is used as an initial reactant; an amino group is used for replacing a 4-bite chlorine atom; an artificial antigen is prepared by using a 1'1-carbonyl diimidazole method; a rabbit-resistant chlorothalonil polyclonal antibody can be obtained by an immune New Zealand white rabbit; the antiserum titer can be determined to be 1:5.12*10<4> by using the ELISA; and on the basis, the residual chlorothalonil ELISA detection method can be established; and the lowest detection concentration by using the method is 0.383ng/mL.

Description

technical field [0001] The invention relates to a novel preparation method of bactericide chlorothalonil antigen antibody and its residue ELISA detection method. The method can quickly and efficiently detect chlorothalonil pesticide residues in fruits and vegetables, and the cost is low. The invention belongs to the field of pesticide residue enzyme immunoassay. Background technique [0002] Chlorothalonil (TCPN), chemical name 2,4,5,6-tetrachloro-1,3-dicyanobenzene, is a broad-spectrum protective fungicide, mainly used in agricultural production to control fruits and vegetables Fungal diseases. Because chlorothalonil has good adhesion to plants and is widely used, chlorothalonil and its metabolites have relatively large residues in fruits and vegetables, soil and water, and can be detected even in arctic regions (Gillian L .Daly, YingD.Lei, Camilla Teixeira, et al.Pesticides in Western Canadian Mountain Air and Soil[J].Environ.Sci.Technol, 2007, 41, 6020-6025.). U.S. En...

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Application Information

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IPC IPC(8): C07K14/765C07K14/77C07K16/44G01N33/543G01N33/52
Inventor 于基成郭乃菲刘秋
Owner DALIAN NATIONALITIES UNIVERSITY
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