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Method for preparing anti-coagulation raticide bromadiolone half antigen and complete antigen

A technology of complete antigen and bromidilone, which is applied in the preparation methods of peptides, chemical instruments and methods, serum albumin and other directions, can solve the problems of many synthesis steps, difficulty in product separation and purification, and inability to prepare antibodies, and achieves the preparation method. Simple and broad application prospects

Inactive Publication Date: 2013-01-30
INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the many synthetic steps in the hapten synthesis process of some poisonous substances, the separation and purification of products are difficult, and sometimes even satisfactory antibodies cannot be prepared.
There is no related literature report about the preparation method of bromadiolone hapten

Method used

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  • Method for preparing anti-coagulation raticide bromadiolone half antigen and complete antigen
  • Method for preparing anti-coagulation raticide bromadiolone half antigen and complete antigen

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0029] The preparation method of the anticoagulant mouse poison bromadiolone hapten of the present embodiment comprises the following steps:

[0030] (1) Add bromadiolone and anhydrous potassium carbonate to 1,4-dioxane respectively, the ratio of bromadiolone to anhydrous potassium carbonate is 1:0.5, bromadiolone is 1, The concentration in 4-dioxane is 0.7mol / L, and the reaction is carried out under the condition of stirring for 20h at a temperature of 15°C; then adding succinic anhydride, the ratio of the amount of bromadiolone to succinic anhydride is 1: 1.5, heating to reflux for 6h;

[0031] (2) Pour the reaction mixture obtained in step (1) into water at 15°C, and off-white solids precipitate out;

[0032] (3) Filtration and ethanol recrystallization to obtain bromadiolone hapten, the yield of bromadiolone hapten is 78%.

Embodiment 2

[0034] The preparation method of the anticoagulant mouse poison bromadiolone hapten of the present embodiment comprises the following steps:

[0035] (1) Add bromadiolone and anhydrous potassium carbonate to 1,4-dioxane respectively, the ratio of bromadiolone to anhydrous potassium carbonate is 1:0.5, bromadiolone is 1, The concentration in 4-dioxane is 0.7mol / L, and the reaction is carried out under the condition of stirring for 20h at a temperature of 15°C; then adding succinic anhydride, the ratio of the amount of bromadiolone to succinic anhydride is 1: 1.5, heating to reflux for 6h;

[0036] (2) Pour the reaction mixture obtained in step (1) into water at 5°C, and off-white solids precipitate out;

[0037] (3) Filtration and ethanol recrystallization to obtain bromadiolone hapten, the yield of bromadiolone hapten is 75%.

Embodiment 3

[0039] The preparation method of the anticoagulant mouse poison bromadiolone hapten of the present embodiment comprises the following steps:

[0040] (1) Add bromadiolone and anhydrous potassium carbonate to 1,4-dioxane respectively, the ratio of bromadiolone to anhydrous potassium carbonate is 1:0.5, bromadiolone is 1, The concentration in 4-dioxane is 0.7mol / L, and the reaction is carried out under the condition of stirring for 20h at a temperature of 15°C; then adding succinic anhydride, the ratio of the amount of bromadiolone to succinic anhydride is 1: 1.5, heating to reflux for 6h;

[0041] (2) Pour the mixture obtained from the reaction in step (1) into the ice-water mixture, and off-white solids precipitate out;

[0042] (3) Filtration and ethanol recrystallization to obtain bromadiolone hapten, the yield of bromadiolone hapten was 83%.

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Abstract

The invention relates to a method for preparing an anti-coagulation raticide bromadiolone half antigen and a complete antigen. The method comprises the following steps of: (1) adding bromadiolone and anhydrous potassium carbonate into 1,4-dioxane respectively, reacting while stirring, adding succinic anhydride, heating and refluxing; (2) pouring a mixture obtained by reacting in the step (1) into water, and precipitating a greyish-white solid out; (3) filtering, and recrystallizing with ethanol to obtain a bromadiolone half antigen; (4) adding the bromadiolone half antigen, dicyclohexylcarbodiimide and BSA (Bovine Serum Albumin) into dimethylsulfoxide, adding a phosphate buffer solution of which the pH is 6-7, and stirring; and (5) dialyzing, and performing freeze drying to obtain a bromadiolone complete antigen. The preparation method provided by the invention is easy, convenient and rapid, can be applied to immune animals, can be used for preparing a monoclonal antibody of bromadiolone and establishing a rapid detection method of anti-coagulation raticide bromadiolone, and is widely applied in the fields of forensic science, clinical medicine and the like.

Description

technical field [0001] The invention relates to a method for preparing a hapten and a complete antigen in the technical field of immunology, in particular to a method for preparing an anticoagulant mouse drug bromadiolone hapten and a complete antigen. Background technique [0002] Bromadiolone is a slow-acting rodenticide, which is a coumarin compound. Using this type of compound can destroy the prothrombin in the mouse body, causing large-scale bleeding in various organs and mucous membranes in the body, and finally achieve the purpose of killing the mouse. , so copper is called an anticoagulant rodenticide. In recent years, acute rodenticides such as tetramine and fluoroacetamide have been banned, and these new-generation anticoagulant rodenticides have been replaced. [0003] Although this type of rodenticide has a good rodent control effect, along with its wide application in recent years, cases and incidents caused by misuse, suicide, etc. have occurred from time to t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D311/56C07K14/765C07K1/107
Inventor 董颖栾玉静张蕾萍王瑞花杜鸿雁王炯常靖于忠山何毅王芳琳侯小平张云峰姚伊人
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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