Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Sulfadimidine molecular imprinting bionic recognition kit, and preparation method and application thereof

A technology for sulfamethazine and sulfamethazine enzyme labeling substances, applied in the field of analytical chemistry, can solve the problems of cumbersome preparation process of specific antibodies, long cycle, poor tolerance to environmental conditions, etc., and achieves easy promotion and popularization, High accuracy and low cost effect

Inactive Publication Date: 2013-09-11
HUAZHONG AGRI UNIV
View PDF6 Cites 14 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional ELISA method is based on biological antibodies as recognition elements, and the preparation process of specific antibodies is cumbersome, the cycle is long and the tolerance to external environmental conditions is poor.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Sulfadimidine molecular imprinting bionic recognition kit, and preparation method and application thereof
  • Sulfadimidine molecular imprinting bionic recognition kit, and preparation method and application thereof
  • Sulfadimidine molecular imprinting bionic recognition kit, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1. Preparation of Sulfamethazine Imprinted Microspheres

[0053] Weigh 0.5mmol of the template molecule (sulfamethazine) and dissolve it in 30mL of acetonitrile, add 3mmol of the functional monomer (MAA), and place it in a refrigerator at 4°C for 24h, so that the monomer and the template molecule can fully interact to complete the prepolymerization process. Then add 15mmol of cross-linking agent EDGMA and 0.3mmol of initiator AIBN in turn, transfer the mixed solution into a 18mm×180mm quartz test tube, pass nitrogen gas for 5min, seal the test tube under nitrogen atmosphere and use 365nm, 6w ultraviolet light in an ice-water bath at 0°C The lamp initiates polymerization, the distance between the lamp tube and the test tube is 3cm, and the polymerization time is 48h. The obtained polymer is centrifuged and eluted with a mixed solvent of methanol-hydrochloric acid (7:3, V / V) until there is no template molecule in the eluent during ultraviolet scanning. UV absorpti...

Embodiment 2

[0059] Embodiment 2. Synthesis of sulfamethazine enzyme marker

[0060] The enzyme label was synthesized by glutaraldehyde coupling (for the synthetic route, see Figure 5 ), the specific method is to accurately weigh 45 mg of HRP, and add 13 mL of 0.05 mol / L pH9.6 carbonate buffer at room temperature (20-25 °C, the same below) to fully dissolve it. Then accurately weigh 10 mg of sulfamethazine and dissolve it in 1 mL of 1 mmol / L sodium hydroxide solution. Take 50 μL of sulfamethazine solution and mix it with HRP solution. Under magnetic stirring, add 15 μL of glutaraldehyde solution with a volume ratio of 25% dropwise for 30 minutes, adjust the pH value to about 9.6 with 1 mmol / L HCL, and continue the reaction for 1 hour. Finally, the ligation product was put into a dialysis bag and dialyzed with 0.01mol / L pH7.4PBS for 5 days. Carry out ultraviolet scanning to the obtained conjugate, carry out qualitative analysis by comparing the change of HRP before and after cross-linkin...

Embodiment 3

[0061] Embodiment 3. Sample pretreatment method

[0062] Extraction of sulfamethazine from pig muscle and liver: Weigh 5g homogenized sample into a 50mL centrifuge tube, add sulfamethazine standard substance to make the concentrations reach 50μg / kg, 100μg / kg and 200μg / kg respectively, and ultrasonically After mixing for 5 minutes, add 2g of anhydrous sodium sulfate, then add 20mL of dichloromethane, vortex extract for 10min, then centrifuge at 3000r / min for 5min, filter the supernatant into a 100mL concentration bottle through quantitative filter paper, and wash the residue in a 15mL dichloromethane water bath Ultrasonic extraction was performed once (15 min), combined with dichloromethane, and blown dry with nitrogen in a water bath at 35°C.

[0063] The above residue was dissolved in 1 mL of methanol water with a volume ratio of 25%, vortexed to mix, transferred to a 5 mL centrifuge tube, added 3 mL of n-hexane, mixed, left to stand for layers, and the upper n-hexane layer w...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Decomposition temperatureaaaaaaaaaa
Login to View More

Abstract

The invention discloses a sulfadimidine molecular imprinting bionic recognition kit, and a preparation method and an application thereof. The kit contains a sulfadimidine molecular imprinting polymer microsphere, a sulfadimidine enzyme-labeled object and a sulfadimidine standard substance solution, wherein the sulfadimidine molecular imprinting polymer microsphere is a bionic antibody which is synthesized by a precipitation polymerization method and has a recognition function to the sulfadimidine; and the sulfadimidine enzyme-labeled object is a compound obtained by coupling sulfadimidine and horse radish peroxidase by means of glutaraldehyde. The preparation method comprises the following steps of: A. preparing the sulfadimidine molecular imprinting polymer microsphere; B. synthesizing the sulfadimidine enzyme-labeled object; C. spreading the sulfadimidine molecular imprinting polymer microsphere on a 96-well polrvinyl benzene ELISA plate by using the adhesive with certain concentration; D. preparing a sulfadimidine molecular imprinting SPE (Solid Phase Extraction) column. The kit is used for analyzing and determining sulfadimidine residuals in animal derived food, is low in cost, convenient to operate, high in accuracy and strong in adaptability.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and specifically relates to a sulfamethazine molecular imprinting bionic recognition kit, and also relates to a preparation method of a sulfamethazine molecular imprinting bionic recognition kit, and also relates to a sulfamethazine molecular imprinting bionic recognition kit Application of identification kits in the analysis of animal food residues. Background technique [0002] Sulfamethazine (Sulfamethazine, SM 2 ) is a chemical antibacterial drug used to prevent bacterial and protozoan infections. This type of drug is a slow antibacterial agent, which can produce adverse reactions such as obvious nephrotoxicity, bone marrow suppression, and rash. Due to its low price and wide antibacterial spectrum, it is widely used in animal husbandry at present, but its abuse and misuse are gradually increasing, resulting in more and more drug residues in animal foods. It's getting serious. In order...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N33/545
Inventor 袁宗辉李兆周彭大鹏王玉莲陶燕飞陈冬梅黄玲利戴梦红刘振利
Owner HUAZHONG AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com