Method for preparing electrochemical biosensor based on horse radish peroxidase-porous zirconium phytate nanoparticle modified glassy carbon electrode

A technology of horseradish peroxidase and biosensor, which is applied in the field of preparation of electrochemical biosensors, can solve the problems of hydrogen peroxide biosensors that have not yet been detected, and achieves low cost, good bio-affinity, and simple preparation method Effect

Inactive Publication Date: 2011-02-02
SHANGHAI NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] So far, there is no biosensor for detecting hydrogen peroxide prepared by porous nano-zirconium phytate at home and abroad.

Method used

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  • Method for preparing electrochemical biosensor based on horse radish peroxidase-porous zirconium phytate nanoparticle modified glassy carbon electrode
  • Method for preparing electrochemical biosensor based on horse radish peroxidase-porous zirconium phytate nanoparticle modified glassy carbon electrode
  • Method for preparing electrochemical biosensor based on horse radish peroxidase-porous zirconium phytate nanoparticle modified glassy carbon electrode

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Preparation method of horseradish peroxidase (HRP) biosensor based on porous zirconium phytate nanoparticles: 0.001mol / L ZrOCl 2 Acidic solution, slowly added dropwise to 0.001mol / L sodium phytate solution heated to 90°C under constant stirring (sodium phytate and ZrOCl 2 The volume ratio of the solution is 1:3), and reacted for 1 h. After cooling, centrifuge (7000rpm), wash with ultrapure water (>18MΩ*cm), and then disperse to obtain nanoporous zirconium phytate colloid; take 3 μL of zirconium phytate micellar dispersion and drop it on the inverted clean glassy carbon electrode surface, put it in the refrigerator (4°C) to dry; then drop 3 μL of 5mg / mL horseradish peroxidase (HRP) solution (pH=7.4PBS) onto the glassy carbon electrode modified by nano-zirconium phytate, and put it in the refrigerator Dry it inside (4°C); finally, drop 3 μL of 1% perfluorosulfonic acid (Nafion) solution on the surface of the modified electrode, and put it in the refrigerator (4°C) to dry...

Embodiment 2

[0035]Preparation method of horseradish peroxidase (HRP) biosensor based on porous zirconium phytate nanoparticles: 0.001mol / L ZrOCl 2 Acidic solution, slowly added dropwise to 0.001mol / L sodium phytate solution heated to 80°C under constant stirring (sodium phytate and ZrOCl 2 The volume ratio of the solution is 1:3), and reacted for 1 h. After cooling, centrifuge (6000rpm), wash with ultrapure water (>18MΩ*cm), and then disperse to obtain nanoporous zirconium phytate colloid; take 3 μL of zirconium phytate micellar dispersion and drop it on the inverted clean glassy carbon electrode surface, put it in the refrigerator (4°C) to dry; then drop 3 μL of 5mg / mL horseradish peroxidase (HRP) solution (pH=7.4PBS) onto the glassy carbon electrode modified by nano-zirconium phytate, and put it in the refrigerator Dry it inside (4°C); finally, drop 3 μL of 1% perfluorosulfonic acid (Nafion) solution on the surface of the modified electrode, and put it in the refrigerator (4°C) to dry ...

Embodiment 3

[0037] Preparation method of horseradish peroxidase (HRP) biosensor based on porous zirconium phytate nanoparticles: 0.001mol / L ZrOCl 2 Acidic solution, slowly added dropwise to 0.001mol / L sodium phytate solution heated to 100°C under constant stirring (sodium phytate and ZrOCl 2 The volume ratio of the solution is 1:3), and reacted for 1 h. After cooling, centrifuge (5000rpm), wash with ultrapure water (>18MΩ.cm), and then disperse to obtain nanoporous zirconium phytate colloid; take 3 μL of zirconium phytate micellar dispersion and drop it on the inverted clean glassy carbon electrode surface, put it in the refrigerator (4°C) to dry; then drop 3 μL of 5mg / mL horseradish peroxidase (HRP) solution (pH=7.4PBS) onto the glassy carbon electrode modified by nano-zirconium phytate, and put it in the refrigerator Dry it inside (4°C); finally, drop 3 μL of 1% perfluorosulfonic acid (Nafion) solution on the surface of the modified electrode, and put it in the refrigerator (4°C) to dr...

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Abstract

The invention discloses a method for preparing an electrochemical biosensor based on a horse radish peroxidase-porous zirconium phytate nanoparticle modified glassy carbon electrode. The method comprises the following steps of: dripping a ZrOCl2 acidic solution into a sodium phytate solution and reacting for 1h; cooling and then centrifuging, and dispersing by using ultrapure water to prepare nano porous zirconium phytate colloid; dripping an appropriate amount of zirconium phytate micelle dispersing solution on the surface of a polished glassy carbon electrode and airing; dripping a horse radish peroxidase solution on a nano zirconium phytate modified glassy carbon electrode, and airing; and dripping an appropriate amount of perfluorinated sulfonic acid solution, and airing to obtain theglassy carbon electrode. The invention has the advantages that: the zirconium phytate colloid dispersing solution with the porous structure is prepared and to be used for fixing biological enzymes, keeps the bioactivity of the enzymes and realizes the direct electron transfer between the enzymes and the electrode; the preparation method is simple; and the response time to the hydrogen peroxide detection is shorter, the detection limit is low, and the repeatability and the stability are good.

Description

technical field [0001] The invention relates to an electrochemical biosensor, in particular to a preparation method of an electrochemical biosensor based on horseradish peroxidase-porous phytic acid zirconium nanoparticles modified glassy carbon electrodes. Background technique [0002] The immobilization of biologically active enzymes is a necessary step in the preparation of biosensors. Choosing a suitable immobilization material is one of the key factors to improve the performance of biosensors and plays a decisive role in the performance of the sensors. Therefore, in the past period of time, people have done a lot of work to find suitable materials to immobilize biological enzymes and proteins. These materials must be able to ensure that the protein is not deactivated, aggregated, and most importantly, the structure and properties of the protein are not damaged during the immobilization process. In recent years, the application of nanomaterials in the field of biosensor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48G01N27/30
Inventor 杨海峰陈希尧王娜
Owner SHANGHAI NORMAL UNIVERSITY
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