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Rapid detection kit for simultaneously detecting sulfonamide, florfenicol, tilmicosin and trimethoprim in pork

A florfenicol and trimethoprim technology, applied in the field of veterinary drug residue analysis and detection, can solve the problems of long detection time, inability to meet veterinary drug residue monitoring, and large sample volume requirements, and achieve high sensitivity and low detection limit , the effect of simple operation

Pending Publication Date: 2021-09-03
NEW HOPE LIUHE +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these traditional methods have disadvantages such as large sample volume requirements and long detection time, so they cannot meet the requirements of monitoring veterinary drug residues before pigs enter the factory

Method used

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  • Rapid detection kit for simultaneously detecting sulfonamide, florfenicol, tilmicosin and trimethoprim in pork
  • Rapid detection kit for simultaneously detecting sulfonamide, florfenicol, tilmicosin and trimethoprim in pork
  • Rapid detection kit for simultaneously detecting sulfonamide, florfenicol, tilmicosin and trimethoprim in pork

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] The synthesis of embodiment 1 florfenicol artificial antigen

[0083] 1. Structural modification of florfenicol hapten

[0084] (1) Rinse the 50ml round bottom flask, dry it with ethanol, fix it on the stirrer, and add a stirring bar;

[0085] (2) Weigh 500mg of florfenicol original drug, add 6ml of pyridine, stir until completely dissolved, reserve the sample, add 153mg of HS, heat to 60 degrees, when the reaction is found to be unchanged by TLC, process, spin dry pyridine, and dissolve with methanol , add silica gel to mix the sample, spin dry the sample and carry out column chromatography, collect the required solution, spin dry, and get 450.2mg of Florfenicol hapten;

[0086] The structural formula of the florfenicol hapten is:

[0087]

[0088] 2. Preparation of Florfenicol Immunogen

[0089] (1) Dissolve 17.08mg of florfenicol hapten in 1.5ml of DMF, add 25.75mg of EDC and 15.46mg of NHS to dissolve, stir at room temperature for 2-3h, and obtain reaction sol...

Embodiment 2

[0096] The synthesis of embodiment 2 sulfa artificial antigens

[0097] 1. Structural modification of sulfonamide hapten

[0098] (1) Weigh 500 mg of sodium hydroxide, dissolve it in an ice bath with 6 ml of water, weigh 600 mg of aminobenzoic acid, add it, stir, add 700 mg of acetanilide sulfonyl chloride in batches, and spot the plate to monitor the reaction;

[0099] (2) After the reaction, the pH was adjusted to about 2, and viscous matter was precipitated, dissolved in ethanol, recrystallized with 70% ethanol, cooled and crystallized overnight, and filtered to obtain 580 mg of the intermediate;

[0100] (3) Dissolve 580mg of intermediate with 8ml of methanol, add 5ml of hydrochloric acid, spot the plate to monitor the reaction, after the reaction is complete, add sodium hydroxide solution to adjust the pH to 7, extract, and spin the collected solution to obtain 430mg of sulfa hapten;

[0101] The structural formula of the sulfonamide hapten is:

[0102]

[0103] 2. P...

Embodiment 3

[0111] Synthesis of embodiment 3 trimethoprim artificial antigen

[0112] 1. Trimethoprim immunogen preparation

[0113] (1) Dissolve 5.19 mg of trimethoprim raw material in 1.5 ml of DMF, stir at 200 rpm for 10 min, add 10.3 mg of EDC, 6.18 mg of NHS to dissolve, stir at room temperature (500 rpm) and activate for 2-3 h to obtain reaction solution 1;

[0114] (2) Weigh 20mg of KLH and dissolve it in 3.5ml of 0.1M sodium bicarbonate solution, stir at 200rpm for 10min to fully dissolve, cool down in an ice bath to 0-4°C, stir at 1000rpm, add the reaction solution 1 dropwise, stir at 500rpm Reaction 24h;

[0115] (3) Put the reaction product into the distilled water and rinse the dialysis bag (10cm), 2L 0.01M PBS, dialysis at 4°C (100rpm) for 3 days, and change the liquid three times a day (one in the morning, one in the middle and one in the evening), for a total of nine times in total. The dialyzed product was centrifuged at 5000rpm for 6min to obtain the trimethoprim immuno...

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Abstract

The invention provides a rapid detection kit for simultaneously detecting sulfanilamide, florfenicol, tilmicosin and trimethoprim in pork, which belongs to the technical field of veterinary drug residue analysis and detection. The kit provided by the invention can efficiently and accurately detect whether the pork contains sulfonamide, florfenicol, tilmicosin and trimethoprim at the same time, and effectively solves the problems of long ELISA reaction time and low colloidal gold sensitivity.

Description

technical field [0001] The invention belongs to the technical field of veterinary drug residue analysis and detection, in particular to a rapid detection kit for simultaneously detecting sulfonamides, florfenicol, tilmicosin and trimethoprim in pork. Background technique [0002] With the development of our country's social economy and the continuous increase of the income of urban and rural residents, people's living standards have been greatly improved. At present, the per capita consumption of pork in our country has reached the international average level. However, while the output of pork products has increased substantially, food safety issues such as veterinary drugs, feed additives, illegal additives and other harmful substances have increasingly become the focus and hot spot of widespread social concern. [0003] At present, the domestic existing detection methods for veterinary drug residues of sulfonamides, florfenicol, tilmicosin and trimethoprim are mainly inst...

Claims

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Application Information

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IPC IPC(8): G01N33/577C07K14/765C07K14/77C07K1/107C07K16/44C07C315/00C07C317/32C07C303/38C07C311/37
CPCG01N33/577G01N33/9446C07K14/765C07K14/77C07K16/44C07C315/00C07C303/38C07K2317/33C07C317/32C07C311/37
Inventor 荣佳亚本勤邢维维刘龙海富新文王雅靖刘薇黄强力金兴李鑫张桂亮刘婷王晨霞
Owner NEW HOPE LIUHE
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