Hapten and antigen of nitrofurans and preparation method and application thereof

A technology of nitrofuran and substances, which is applied in the field of haptens and antigens of nitrofuran substances and their preparation and application. It can solve the problems of complex pretreatment, high technical requirements for operation, and long operation time, and shorten the detection time. , Improve detection efficiency and reduce detection cost

Inactive Publication Date: 2010-02-10
北京维德维康生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, LC-UV, LC-MS and LC-MS / MS are currently the most commonly used methods for detecting metabolites of nitrofurans, but in actual operation, LC-UV, LC-MS and LC-MS are required. / MS and other high-end instruments, and the pre-treatment is complex and cumbersome, the operation time is long, and the operation technology requirements are high

Method used

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  • Hapten and antigen of nitrofurans and preparation method and application thereof
  • Hapten and antigen of nitrofurans and preparation method and application thereof
  • Hapten and antigen of nitrofurans and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Synthesis and identification of embodiment 1, formula I structure compound

[0041] 1, the synthesis of formula I structure compound

[0042] 1) Dissolve 139mg (1mmol) of 5-nitrofuran-2-carbaldehyde in an aqueous solution of ethanol consisting of 1mL of pure water and 1mL of absolute ethanol, and magnetically stir to completely dissolve it;

[0043] 2) Dissolve 179mg (1mmol) p-aminophenylbutyric acid in 1mL absolute ethanol;

[0044] 3) Mix the solution of the above step 1) and step 2), the color of the mixture gradually turns yellow, after stirring for 1 minute, a yellow precipitate appears, continue to stir at room temperature, overnight, and centrifuge to obtain a yellow precipitate; dry the precipitate at 60°C , 223mg of product was obtained by weighing, the yield was 71%.

[0045] 2. Identification of the compound of formula I

[0046] The molecular structure of the compound obtained in the above step 1 was determined by ESI. The result is as figure 2 shown. ...

Embodiment 2

[0050]Embodiment 2, the synthesis and identification of nitrofuran compound

[0051] 1. Synthesis of nitrofuran compounds

[0052] 1) Dissolve 15 mg of the compound of the formula I prepared in Example 1 above in 1 mL of N,N-dimethylformamide, and magnetically stir to completely dissolve it;

[0053] 2) Add 15 mg DCC and 15 mg NHS to the solution of the above step 1) respectively, react at room temperature overnight, and centrifuge to remove the precipitate;

[0054] 3) 50 mg of bovine serum albumin (BSA) was dissolved in 5 mL of 0.1 mol / L sodium carbonate solution of pH 9.6;

[0055] 4) Add the solution of the above step 3) dropwise to the solution of the above step 2), and stir and react for 4 hours;

[0056] 5) Put the reaction solution of the above step 4) into a dialysis bag, dialyze with a normal saline solution for 72 hours at 4°C, and change the dialysate 6 times during this period; pour out the dialysate, and put the solution in the dialysis bag under sterile condit...

Embodiment 3

[0059] Embodiment 3, the preparation of antibody

[0060] 1. Preparation of antiserum from immunized animals

[0061] Five New Zealand white rabbits were used as immunized animals, and the nitrofuran compound prepared in the above-mentioned Example 2 was used as the immunogen, and the immunizing dose was 1 mg / kg. The immunization method is as follows: For the first immunization, fully emulsify the immunogen with an equal volume of Freund’s complete adjuvant, inject it subcutaneously at multiple points on the back of the neck, and mix the immunogen with an equal volume of Freund’s incomplete adjuvant to prepare into an emulsifying agent, booster immunization once, a total of 8 times of immunization (including a total of 8 times of booster immunization), and no adjuvant was added for the last time.

[0062] The rabbits were killed 7-10 days after the last immunization, and blood was collected by heart blood collection. Each rabbit can collect about 80 mL of blood. Place the col...

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Abstract

The invention discloses hapten and antigen of nitrofurans and a preparation method and application thereof. The antigen is a nitrofuran compound which is conjugate obtained by conjugating the compoundof formula I structure and carrier protein. As the antigens of the invention have 5-nitrofuran ring structure in common, the antibodies of the nitrofurans can be obtained by taking the antigens as immunogen to immunize animals. The antibodies can simultaneously identify four kinds of nitrofurans (nitrofurazone, furazolidone, furaltadone and nitrofurantion). Therefore, the antigen prepared by theinvention can be used for immunoassay and screening of the nitrofurans, improve the detection efficiency, shorten the detection time and lower the detection cost.

Description

technical field [0001] The invention relates to a hapten and antigen of nitrofuran substances, a preparation method and application thereof. Background technique [0002] Nitrofurans are a class of synthetic broad-spectrum antibacterial drugs with a basic structure of 5-nitrofuran ring, mainly including Nitrofurazone, Nitrofurantion, Furazolidone and Furazidone ( Furaltadone). Nitrofuran substances have a killing effect on most Gram-positive bacteria, Gram-negative bacteria, fungi, protozoa and other pathogens, and are widely used in livestock and poultry farming and aquaculture. However, nitrofurans have certain toxicity to livestock and poultry, large doses or long-term continuous administration of nitrofurans can easily cause toxic reactions, manifested as anorexia, diarrhea, gastrointestinal bleeding, peripheral neuritis, excitement, convulsions or Paralysis, etc., can even cause animal death when the poisoning is severe. At the same time, nitrofurans are also a class...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D307/71C07K16/44C07K14/765C07K14/77C07K1/113G01N33/566G01N33/531
Inventor 吴小平江海洋王战辉阮永磊王进王亚辉徐飞
Owner 北京维德维康生物技术有限公司
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