Label-free impedance type immunosensor for furaltadone residues, preparation method and application thereof

An immunosensor and furatanone technology, applied in material impedance, instruments, scientific instruments, etc., can solve the problems of heavy detection workload, inability to achieve rapid detection, and long time required, and achieve low pretreatment requirements and detection. The effect of high sensitivity and simple analysis process

Inactive Publication Date: 2011-01-26
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing rapid detection methods for the above residues (ELISA and LC-MS-MS methods) all need to derivatize the residues, s...

Method used

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  • Label-free impedance type immunosensor for furaltadone residues, preparation method and application thereof
  • Label-free impedance type immunosensor for furaltadone residues, preparation method and application thereof
  • Label-free impedance type immunosensor for furaltadone residues, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1 The preparation of sensor

[0024] (1) Preparation of gold colloid

[0025] The preparation of gold colloid can be carried out according to prior art. 1:3 (v / v) HNO is required for all glassware used to prepare nano-gold colloids 3 - Soak in HCl solution for a period of time, rinse with double distilled water before use. The preparation method of gold nanoparticles in this example is as follows: 100 mL of 0.01% HAuCl 4 The solution was heated to boiling, and 2.50 mL of 1.0% sodium citrate solution was added under vigorous stirring. The color of the solution first changed from colorless to black, then dark green, and finally wine red and transparent. The particle size of the prepared gold nanoparticles was about 25 nm according to transmission electron microscope observation. In addition, according to the ultraviolet-visible absorption spectrum curve of the nano-gold colloid solution, the absorption peak of the nano-gold colloid appeared at around 520 ...

Embodiment 2

[0035] The establishment of embodiment two detection method

[0036] (1) Optimization of antibody immobilization conditions

[0037] ①Influence of antibody concentration

[0038] The response of the sensor to the AMOZ antigen is related to the amount of AMOZ antibody immobilized on the electrode surface. The more the amount of antibody immobilized per unit area on the electrode surface, the more sites that can bind to the antigen, which is more conducive to the combination with the antigen molecule. In the preparation of the sensor, when the amount of AMOZ antibody was changed, the performance of the obtained sensor showed obvious differences. Depend on figure 1 It can be seen that when the antibody concentration is less than 0.025 mg / mL, the impedance increases with the increase of antibody concentration; when the antibody concentration is greater than 0.025 mg / mL, the impedance response is basically unchanged. Therefore, the antibody concentration was chosen t...

Embodiment 3

[0050] Example 3 AMOZ Detection

[0051] Prepare different concentrations of 3-amino-5-morpholino-2-oxazolone (AMOZ) standard solutions, and conduct the modified electrode containing the immobilized antibody with different concentrations of AMOZ standard solutions under the optimal conditions obtained from the above optimization. immune response. Then the three-electrode system of modified electrode, saturated calomel electrode and platinum electrode was immersed in 1.0 mM Fe(CN) 6 3- / 4- 0.1M KNO 3 In the impedance test solution of phosphate buffer solution (pH 7.4), use Autolab electrochemical workstation under the condition of open circuit potential and sine wave potential amplitude of 10 mV, at 10 -1 Hz to 10 6 Measure its AC impedance spectrum in the Hz frequency range, and then obtain Fe(CN) through equivalent circuit fitting 6 3- / 4 The impedance value of electron transfer when redox reaction occurs on the surface of the modified electrode, and the data is proc...

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Abstract

The invention relates to a label-free impedance type immunosensor for furaltadone residues, a preparation method and application thereof. The label-free impedance type immunosensor is prepared by 5 steps of preparing gold glue, preprocessing gold electrodes, embellishing the gold electrodes by utilizing 1,4-benzodiazepines mercaptan, self-assembling single-layer nanogold glue and fixing an antibody. The label-free impedance type immunosensor is characterized in that nanogold is fixed on the surface of the gold electrodes through a self-assembling method and used for fixing a monoclonal antibody of the furaltadone residues; the relation of the relative change rate %delta Rct and the concentration of AMOZ of impedance is firstly established when the label-free impedance type immunosensor isused; and the concentration of the AMOZ can be determined according to the relative change rate %delta Rct of the impedance of a sample to be detected. The label-free impedance type immunosensor andthe detection method have the characteristic rapid, simple and convenient operation and solve the defect of time waste in the process of detecting traditional furaltadone residues.

Description

technical field [0001] The invention belongs to the field of detection of drug residues; in particular, it relates to a label-free impedance type immunosensor for furaltadone residues and a preparation method and application thereof. Background technique [0002] Nitrofuran drugs are synthetic broad-spectrum antibiotics, which have very good antibacterial effect and pharmacokinetic properties. In addition to being used as an antibiotic in veterinary medicine, it has also been used as a growth-promoting agent for pigs, poultry and aquatic products. additives are widely used. However, in the course of long-term laboratory research, it was found that nitrofuran drugs and their metabolites can cause cancer and gene mutations in experimental animals. (EU Directive 1442 / 95) legislation to prohibit the use of furaltadone in the production and processing of food of animal origin ( Furaltadone ), nitrofurazone ( Nitrofurazone ), furanidine ( Nitrofurantoin ) and furazolidone ( ...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N27/02G01N27/30
Inventor 金文杰杨功俊秦爱建吴丽萍王倩倩邵红霞钱琨余兵李东明蔡宝亮
Owner YANGZHOU UNIV
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