Preparation method of anticoagulant raticide brodifacoum hapten and holoantigen

A complete antigen and hapten technology, applied in the preparation methods of peptides, chemical instruments and methods, serum albumin, etc., can solve the problems of many synthesis steps, difficult separation and purification of products, inability to prepare antibodies, etc., and achieve a simple preparation method. , the effect of broad application prospects

Inactive Publication Date: 2014-12-17
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 relevant literature report on the preparation method of Dalong hapten and complete antigen

Method used

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  • Preparation method of anticoagulant raticide brodifacoum hapten and holoantigen
  • Preparation method of anticoagulant raticide brodifacoum hapten and holoantigen
  • Preparation method of anticoagulant raticide brodifacoum hapten and holoantigen

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The preparation method of the anticoagulant rat poison Dalong hapten in this embodiment comprises the following steps:

[0030] (1) Add Dalong and succinic anhydride to tetrahydrofuran respectively, the ratio of the amount of Dalong to succinic anhydride is 1:1, the concentration of Dalong in THF is 0.1mol / L, at a temperature of 30 Stir for 5h under the condition of ℃ (the reaction is shown in formula I, and the embodiment 2-6 is the same);

[0031]

[0032] (2) Pour the reaction mixture obtained in step (1) into water at 10°C, and off-white solids precipitate out;

[0033] (3) Filtration and ethanol recrystallization to obtain the Dalong hapten with a yield of 85.6%.

Embodiment 2

[0035] The preparation method of the anticoagulant rat poison Dalong hapten in this embodiment comprises the following steps:

[0036] (1) Add Dalong and succinic anhydride to tetrahydrofuran respectively, the ratio of the amount of Dalong to succinic anhydride is 1:1, the concentration of Dalong in THF is 0.1mol / L, at a temperature of 30 Stirring at ℃ for 5h;

[0037] (2) Pour the reaction mixture obtained in step (1) into water at 5°C, and off-white solids precipitate out;

[0038] (3) Filtration and ethanol recrystallization to obtain the Dalong hapten with a yield of 82.4%.

Embodiment 3

[0040] The preparation method of the anticoagulant rat poison Dalong hapten in this embodiment comprises the following steps:

[0041] (1) Add Dalong and succinic anhydride to tetrahydrofuran respectively, the ratio of the amount of Dalong to succinic anhydride is 1:1, the concentration of Dalong in THF is 0.1mol / L, at a temperature of 30 Stirring at ℃ for 5h;

[0042] (2) Pour the mixture obtained from the reaction in step (1) into the ice-water mixture, and off-white solids precipitate out;

[0043] (3) Filtration and ethanol recrystallization to obtain Dalong hapten with a yield of 91.2%.

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Abstract

The invention relates to a preparation method of anticoagulant raticide brodifacoum hapten and holoantigen, which comprises the following steps: (1) respectively adding brodifacoum and succinic anhydride into tetrahydrofuran to carry out reaction while stirring; (2) pouring the mixture obtained by the reaction in the step (1) into water to precipitate an offwhite solid; (3) filtering, and recrystallizing with ethanol to obtain the brodifacoum hapten; (4) respectively adding the brodifacoum hapten, dicyclohexylcarbodiimide and BSA (bovine serum albumin) into dimethyl sulfoxide, adding a phosphoric acid buffer solution with the pH value of 6-7, and stirring; and (5) dialyzing, and carrying out freeze-drying to obtain the brodifacoum holoantigen. The preparation method provided by the invention is simple and quick, can be used for immune animals, and can be used for preparing monoclonal antibodies of brodifacoum, thereby establishing a quick detection method of anticoagulant raticide brodifacoum. The invention is widely applicable to 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 rat drug Dalong hapten and a complete antigen. Background technique [0002] Dalong 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. Therefore, copper is called 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 time. This ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D311/56C07K14/765C07K1/107
Inventor 董颖杜鸿雁栾玉静王炯王瑞花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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