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Method for inhibiting peroxidase activity by using fluorescent graphene quantum dots

A technology of graphene quantum dots and peroxidase, which is applied in the direction of material analysis by observing the influence on chemical indicators, color/spectral characteristic measurement, and analysis by making materials undergo chemical reactions, which can solve the problems of poor dispersion, Interfering with enzyme activity, easy restriction enzyme structure opening and other problems, to achieve wide application and solve performance effects

Inactive Publication Date: 2015-03-25
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved in the present invention is to provide a new enzyme regulator for the existing graphite oxide as an enzyme regulator, which has poor dispersibility and a micron size, which is easy to limit the opening of the enzyme structure and interfere with the enzyme activity. Preparation method and use for regulating enzyme activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028]Add 4g of citric acid and 1g of glycine to 5mL of water to prepare a mixed solution, heat the mixed solution until the water evaporates completely, transfer it to an autoclave, and heat it to 220°C for 3 hours. After the reaction, the product was dissolved in a certain volume of water, the pH value of the system was adjusted to neutral with 1M sodium hydroxide, a homogeneous solution was obtained after dialysis with a dialysis bag (molecular weight cut-off 1000Da), and solid graphene quantum dots were obtained after drying. The obtained graphene quantum dots are dispersed in double distilled water to form a stable graphene quantum dot solution with a concentration of 750 μg / mL. Take 200 μL of this quantum dot solution and 100 μL of 200ng / mL horseradish peroxidase to add 4 mL of pH value 4 in the acetic acid buffer solution, placed at room temperature for 30 min, added 200 μL of 5 mM chromogenic agent tetramethylbenzidine, 500 μL of 1 mM H 2 o 2 Standard solution, shake ...

Embodiment 2

[0030] Add 4g of citric acid and 2g of lysine to 5mL of water to prepare a mixed solution, heat the mixed solution until the water evaporates completely, then transfer it to an autoclave, and heat it to 200°C for 3 hours. After the reaction, the product was dissolved in a certain volume of water, the pH value of the system was adjusted to neutral with 1M sodium hydroxide, a homogeneous solution was obtained after dialysis with a dialysis bag (molecular weight cut-off 1000Da), and solid graphene quantum dots were obtained after drying. The obtained graphene quantum dots are dispersed in double-distilled water to form a stable graphene quantum dot solution with a concentration of 200 μg / mL. Take 200 μL of this quantum dot solution and 100 μL of 500ng / mL horseradish peroxidase to add 4 mL of pH value 9 in phosphate buffer solution, placed at room temperature for 2 hours, added 200 μL 0.5 mM chromogenic agent tetramethylbenzidine, 500 μL 1 mM H 2 o 2 Standard solution, shake the ...

Embodiment 3

[0032] Add 2g of citric acid and 1g of glycine to prepare a mixed solution by adding 5mL of water, heat the mixed solution until the water is completely evaporated, transfer it to an autoclave, and heat it to 250°C for 2 hours. After the reaction, the product was dissolved in a certain volume of water, the pH value of the system was adjusted to neutral with 1M sodium bicarbonate, a uniform solution was obtained after dialysis with a dialysis bag (molecular weight cut-off 1000Da), and solid graphene quantum dots were obtained after drying. The obtained graphene quantum dots are dispersed in double-distilled water to form a stable graphene quantum dot solution with a concentration of 250 μg / mL. Take 200 μL of this quantum dot solution and 100 μL of 500ng / mL horseradish peroxidase to add 4 mL of pH value 4 in the acetic acid buffer solution, placed at room temperature for 1.5h, added 200μL 5mM color reagent diaminobenzidine, 500μL 0.5mM H 2 o 2 Standard solution, shake the mixed...

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Abstract

The invention provides a method for inhibiting peroxidase activity by using fluorescent graphene quantum dots. The method comprises the following steps: (1) adding a carbon source into a certain amount of water to prepare a mixed solution, heating the solution until the moisture is completely evaporated, transferring the solution into a high pressure reactor, heating and reacting for a period of time, dissolving the product in water, regulating the pH value of the system to be neutral by using alkali liquor, purifying the product, and drying, thereby obtaining solid graphene quantum dots; and (2) dispersing the quantum dots obtained in the step (1) in secondary distilled water, taking a proper amount of quantum dot solution and enzyme solution, adding the solution into a buffer solution, adding a color developing agent and H2O2 after a period of time, uniformly shaking the mixed solution, rapidly adding the solution into a cuvette, measuring the light absorption value of the maximum absorption wavelength of the product on an ultraviolet-visible spectrophotometer, and performing blank sample control, thereby determining the relative enzyme activity. The graphene quantum dots are prepared by adopting low-price and readily available raw materials, and the method is green and environment-friendly. The structure of the enzyme can be changed by virtue of the interaction between the graphene quantum dots and zymoprotein, and regulation of the enzyme activity is realized.

Description

technical field [0001] The invention belongs to the field of chemical industry, in particular to a method for inhibiting peroxidase activity by fluorescent graphene quantum dots. Background technique [0002] Nanomaterials are materials with at least one dimension in the nanometer scale. When they are in the nanometer scale, some properties of the material will also change significantly, and at the same time, they will have some special properties different from ordinary materials, such as small size Effect, Quantum Confinement Effect, Macroscopic Quantum Tunneling Effect, Surface Effect, etc. These special effects endow nanomaterials with unique physical, chemical, electrical, mechanical and thermal properties, and have been widely used in various fields. In recent years, research on the application of nanomaterials in the medical field has also received close attention. As nanomaterials show more and more broad application prospects in the field of biomedicine, it is par...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/31G01N21/33
Inventor 李在均周晓燕
Owner JIANGNAN UNIV
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