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Enzyme electrode for detecting sterigmatocystin and preparation and application thereof

A variegated aspergillus and enzyme electrode technology, which is applied in the field of biosensors, can solve the problems that the detection sensitivity needs to be improved, the stability and reproducibility need to be improved, etc., and achieves rapid and convenient detection, good alkaline environment tolerance, Use simple effects

Inactive Publication Date: 2011-09-07
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection sensitivity of this type of enzyme biosensor needs to be improved, the stability and reproducibility need to be improved, and the detection potential can be further optimized to improve the selectivity and anti-interference ability of the enzyme electrode.

Method used

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  • Enzyme electrode for detecting sterigmatocystin and preparation and application thereof
  • Enzyme electrode for detecting sterigmatocystin and preparation and application thereof
  • Enzyme electrode for detecting sterigmatocystin and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Preparation of an enzyme biosensor for detecting ST

[0031] (1) Instruments and reagents

[0032] CHI660C electrochemical workstation (Shanghai Chenhua Instrument Co., Ltd.), the experiment uses a three-electrode system: a gold electrode (inner diameter of 2 mm) or a modified electrode is used as the working electrode; an Ag / AgCl (saturated KCl) electrode is used as the reference electrode; platinum wire The electrodes are auxiliary electrodes.

[0033] Chitosan (CS, 85-90% degree of deacetylation, average molecular weight 1 x 10 6 g / mol, Zhejiang Aoxing Biotechnology Co., Ltd.); single-walled carbon nanotubes (SWCNTs, diameterm, purity>95%, Shenzhen Nanoport Company), L-cysteine (L-Cys, >99%, Beijing Qihuasheng Biotechnology Development Center), Potassium Ferricyanide (Guangzhou Chemical Reagent Factory), Ferric Chloride (Guangzhou Chemical Reagent Factory), Dipotassium Hydrogen Phosphate (Guangzhou Chemical Reagent Factory) ), potassium dihydrogen phos...

Embodiment 2

[0048] Example 2: Study on the alkaline environment stability of the hybrid Prussian blue modified electrode

[0049] Using CHI660C electrochemical workstation (Shanghai Chenhua Instrument Co., Ltd.), using a three-electrode system (reference electrode: Ag / AgCl (saturated KCl) electrode; counter electrode: platinum wire electrode; working electrode: Prussian blue modified electrode), select the cycle voltage In the scanning mode, set the electrochemical parameters as follows:

[0050] Working potential: -0.05V~+0.35V

[0051] Scan rate: 50mV / s

[0052] Rest time: 2s

[0053] After setting the parameters, in the pH8.0 supporting electrolyte PBS buffer (0.05 mol / L KH 2 PO 4 -K 2 HPO 4 + 0.1 mol / L KCl), the current response signal of the hybrid Prussian blue modified electrode (PB-CS / Cys / Au) was detected by cyclic voltammetry. For comparison, a pure Prussian blue modified electrode (PB / Au) was assembled at the same time, and the assembly process was similar to the hybrid ...

Embodiment 3

[0054] Example 3: Electrochemical response of CS-AFO-SWCNTs / PB-CS / Cys / Au modified electrode to ST

[0055] (1) Cyclic voltammetry detection of ST:

[0056]Using CHI660C electrochemical workstation (Shanghai Chenhua Instrument Co., Ltd.), using a three-electrode system (reference electrode: Ag / AgCl (saturated KCl) electrode, counter electrode: platinum wire electrode, working electrode: CS-AFO-SWCNTs / PB-CS / Cys / Au modified electrode), select the cyclic voltammetry scan mode, and set the electrochemical parameters as follows:

[0057] Working potential: -0.05V~+0.35V

[0058] Scan rate: 50mV / s

[0059] Rest time: 2s

[0060] After setting the parameters, the pH6.5 supporting electrolyte PBS buffer (0.05 mol / L KH 2 PO 4 -K 2 HPO 4 + 0.1 mol / L KCl) were added dropwise with different concentrations of ST, mixed and detected by cyclic voltammetry. Such as figure 2 As shown, in PBS without ST, the CS-AFO-SWCNTs / PB-CS / Cys / Au modified electrode exhibited a pair of redox peak...

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Abstract

The invention relates to an enzyme electrode for detecting sterigmatocystin and a preparation method thereof, and a biosensor which uses the enzyme electrode as a substrate electrode for fixing and assembling aflatoxin oxidase and is used for detecting sterigmatocystin. An L-cysteine functional film is self-assembled on the surface of a gold electrode, and then the gold electrode is immersed into heterozygous Prussian-blue electrode-position working solution to prepare the Prussian-blue heterozygous electrode by a potentiostatic method. CNT (carbon nano tubes) used as electron carriers are evenly dispersed in chitosan solution and are used as carriers for embedding aflatoxin oxidase. Then the chitosan- aflatoxin oxidase-CNT mixed film is assembled on the surface of the Prussian-blue heterozygous modified electrode, and finally, the biosensor used for detecting sterigmatocystin is obtained. The enzyme electrode and the biosensor provided by the invention have better stability and repeatability, quick response to sterigmatocystin, low voltage detection, strong antijamming capacity and excellent selectivity.

Description

technical field [0001] The invention belongs to the field of biosensors, and relates to an enzyme electrode, a biosensor assembled from the enzyme electrode, and applications of the enzyme electrode and the biosensor. Background technique [0002] Prussian blue (Prussian blue, PB) is an effective redox mediator. The PB film modified electrode has excellent electrochemical reversibility, high stability and electrocatalytic performance. 2 o 2 With highly sensitive electrocatalysis, it is called "artificial peroxidase". PB modified electrode can effectively reduce the detection of H 2 o 2 The overpotential of the solution avoids the interference of the coexisting substances in the solution on the determination. Amperometric oxidase biosensors usually detect enzymatic reaction product H 2 o 2 To achieve the purpose of detecting the substrate, therefore, the oxidase biosensor based on PB modified membrane assembly has developed rapidly in clinical, environmental and food an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/403
Inventor 刘大岭陈俊华姚冬生谢春芳
Owner JINAN UNIVERSITY
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