Enzyme biosensor for detecting cholesterol as well as preparation method and application of biosensor

A biosensor, cholesterol oxidase technology, applied in the field of electrochemical biosensors, to achieve good electron transfer, good embedding effect, and good selectivity

Inactive Publication Date: 2014-07-30
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, graphene, a carbon nanomaterial with excellent physical and chemical properties, has a large specific surface area, excellent electrical conductivity and carrier properties. Thionine is a widely used electron mediator with the ability to quickly transfer electrons. Both have been applied to the preparation of glucose and hydrogen peroxide biosensors, but the application of cholesterol biosensing has not yet been reported

Method used

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  • Enzyme biosensor for detecting cholesterol as well as preparation method and application of biosensor
  • Enzyme biosensor for detecting cholesterol as well as preparation method and application of biosensor
  • Enzyme biosensor for detecting cholesterol as well as preparation method and application of biosensor

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Experimental program
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Effect test

Embodiment 1

[0040] An enzyme biosensor for detecting cholesterol, the schematic diagram of the overall structure is shown in figure 1 As shown, it consists of a reference electrode 2, a counter electrode 3 and a modified working electrode 1; the cross-sectional schematic diagram of the modified working electrode 1 is shown in figure 2 As shown, the modified working electrode 1 is composed of a working electrode 1-1 and a material recognition enzyme film 1-2 solidified on the surface of the working electrode, wherein the material recognition enzyme film 1-2 is mainly composed of graphene, sulfur Corydalis, cholesterol oxidase, horseradish peroxidase and chitosan are mixed and prepared;

[0041] The substance recognition enzyme membrane 1-2 is obtained by taking 1-5 mg / mL graphene-chitosan solution, 5-10 mmol / L thionine aqueous solution, 20-30 mg / mL cholesterol oxidase aqueous solution and 10-20 mg / L mL of horseradish peroxidase aqueous solution was mixed and then dried;

[0042] Prefera...

Embodiment 2

[0046] A kind of enzyme biosensor for detecting cholesterol, its preparation method is as follows:

[0047] (1) Take 5g flake graphite and 2g NaNO 3 For mixing, add 120mL concentrated sulfuric acid, place in 4°C ice-water bath and stir for 30min; then add 20gKMnO 4 , magnetically stirred and reacted for 60 minutes, then moved into a warm water bath at 40°C and continued to react for 30 minutes; then added 230mL of distilled water, stirred at a temperature of 95°C for 5 minutes, then added hydrogen peroxide with a mass fraction of 5%, until no bubbles were generated, filtered while hot, And use deionized water and 5% hydrochloric acid for multiple washings; then dialyze for one week, then dry in a vacuum oven at 60°C to obtain graphite oxide; weigh 0.1g of the graphite oxide and dissolve it in 50g of distilled water, ultrasonically 60min to obtain a stable graphite dispersion; heat the graphite dispersion to 80°C and add 2mL of hydrazine hydrate dropwise, filter after reacting...

Embodiment 3

[0059] A kind of enzyme biosensor for detecting cholesterol, its preparation method is as follows:

[0060] (1) Take 5g flake graphite and 2g NaNO 3 For mixing, add 120mL concentrated sulfuric acid, place in 4°C ice-water bath and stir for 30min; then add 20gKMnO 4 , magnetically stirred and reacted for 60 minutes, then moved into a warm water bath at 40°C and continued to react for 30 minutes; then added 230mL of distilled water, stirred at a temperature of 95°C for 5 minutes, then added hydrogen peroxide with a mass fraction of 5%, until no bubbles were generated, filtered while hot, And use deionized water and 5% hydrochloric acid for multiple washings; then dialyze for one week, then dry in a vacuum oven at 60°C to obtain graphite oxide; weigh 0.1g of the graphite oxide and dissolve it in 50g of distilled water, ultrasonically 60min to obtain a stable graphite dispersion; heat the graphite dispersion to 80°C and add 2mL of hydrazine hydrate dropwise, filter after reacting...

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Abstract

The invention belongs to the technical field of electrochemical biosensors and particularly relates to an enzyme biosensor for detecting cholesterol as well as a preparation method and an application of the biosensor. The enzyme biosensor adopts a classical three-electrode system, wherein a specific matter recognizing enzyme membrane is solidified on a working electrode and mainly formed by mixing graphene, thionine, cholesterol oxidase, horse radish peroxidase and chitosan. The preparation method of the enzyme biosensor comprises the following steps: firstly uniformly mixing a thionine solution, a chitosan solution of the graphene, a horse radish peroxidase solution and a cholesterol oxidase solution, dripping onto the processed working electrode, forming the three-electrode system together with a reference electrode and a counter electrode to obtain the enzyme biosensor. The enzyme biosensor disclosed by the invention has the advantages of economical efficiency, simplicity, rapidness, sensitivity and the like, is simple in preparation and can be used for the quantitative determination of the cholesterol.

Description

technical field [0001] The invention belongs to the technical field of electrochemical biosensors, and in particular relates to an enzyme biosensor for detecting cholesterol, a preparation method and application thereof. Background technique [0002] Cholesterol is an indispensable and important substance in animal tissue cells. It not only participates in the formation of cell membranes, but also is a raw material for the synthesis of bile acids, vitamin D and steroid hormones. However, as the level of development increases, the incidence of abnormal cholesterol increases. Modern studies have found that heart disease, diabetes, atherosclerosis, venous thrombosis, and cholelithiasis are closely related to hypercholesterolemia. Therefore, rapid determination of cholesterol is of great significance. At present, enzymatic colorimetry based on spectrophotometers is widely used in clinical testing. However, this method uses a large amount of enzymes, complex instruments, long ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/327
Inventor 朴金花张秀花
Owner SOUTH CHINA UNIV OF TECH
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