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Graphite alkyne and click chemistry-based copper ion detection method, kit and application

A detection method and click chemistry technology, applied in the field of detection, can solve the problems of reducing the repeatability of the reaction, increasing the complexity of the reaction, etc., and achieve the effects of low detection limit, high specificity, and simplified operation time and steps.

Active Publication Date: 2020-03-24
SHENZHEN PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, these conventional two-dimensional materials for copper ion detection methods need to modify the two-dimensional materials with additional alkyne ligands, which increases the complexity of the reaction, and the heterogeneity of the two-dimensional material modification reduces the reaction efficiency. Repeatability

Method used

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  • Graphite alkyne and click chemistry-based copper ion detection method, kit and application
  • Graphite alkyne and click chemistry-based copper ion detection method, kit and application
  • Graphite alkyne and click chemistry-based copper ion detection method, kit and application

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Embodiment 1

[0058] The present embodiment provides a kind of detection method based on graphdiyne and the copper ion of click chemistry, comprises the steps:

[0059] (1) Add DNA1 with a concentration of 10 μM and a volume of 10 μL and DNA2 with a concentration of 10 μM and a volume of 10 μL into PBS buffer, mix at 95°C for 5 minutes to obtain group-modified double-stranded DNA, and store at 4°C;

[0060] (2) The group-modified double-stranded DNA obtained in step (1) and 2 μg graphyne were vortexed for 20 s in 0.2 mL of PBS buffer to obtain a mixed solution of graphyne and group-modified double-stranded DNA;

[0061] (3) adding a volume of 20 μL to the mixed solution of graphyne and group-modified double-stranded DNA obtained in step (2) is the copper ion standard solution of 100 nM, 500 nM, 1 μM, 5 μM, 10 μM, 100 μM and 1 mM respectively A volume of 20 μL of sodium ascorbate solution with a concentration of 5 mM was mixed to obtain a mixture (the concentration of graphyne in the mixture...

Embodiment 2

[0079] The present embodiment provides a kind of detection method based on graphdiyne and the copper ion of click chemistry, comprises the steps:

[0080] (1) Add DNA1 with a concentration of 5 μM and a volume of 10 μL and DNA2 with a concentration of 5 μM and a volume of 10 μL into PBS buffer, mix at 80°C for 8 minutes to obtain group-modified double-stranded DNA, and store at 4°C;

[0081] (2) The group-modified double-stranded DNA obtained in step (1) and 2.5 μg graphyne were vortexed and mixed for 30 seconds in 0.5 mL of PBS buffer to obtain a mixture of graphyne and group-modified double-stranded DNA ;

[0082] (3) Adding a volume of 50 μL to the mixed solution of graphyne and group-modified double-stranded DNA obtained in step (2) is the copper ion standard solution of 100 nM, 500 nM, 1 μM, 5 μM, 10 μM, 100 μM and 1 mM respectively and A volume of 50 μL of sodium ascorbate solution with a concentration of 1 mM was mixed to obtain a mixture (the concentration of graphyne...

Embodiment 3

[0092] The present embodiment provides a kind of detection method based on graphdiyne and the copper ion of click chemistry, comprises the steps:

[0093] (1) Add DNA1 with a concentration of 15 μM and a volume of 10 μL and DNA2 with a concentration of 15 μM and a volume of 10 μL into PBS buffer, mix at 100°C for 3 minutes to obtain group-modified double-stranded DNA, and store at 4°C;

[0094] (2) The group-modified double-stranded DNA obtained in step (1) and 7.5 μg graphyne were vortexed and mixed for 10 s in 0.5 mL of PBS buffer to obtain a mixture of graphyne and group-modified double-stranded DNA ;

[0095] (3) Adding a volume of 50 μL to the mixed solution of graphyne and group-modified double-stranded DNA obtained in step (2) is the copper ion standard solution of 100 nM, 500 nM, 1 μM, 5 μM, 10 μM, 100 μM and 1 mM respectively and A volume of 50 μL of sodium ascorbate solution with a concentration of 1 mM was mixed to obtain a mixture (the concentration of graphyne in...

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Abstract

The invention provides a graphdiyne and click chemistry-based copper ion detection method, a kit and an application. The detection method comprises the following steps of (1) adding a copper ion standard solution with a concentration gradient and a reducing agent into a mixed solution of graphdiyne and group-modified double-stranded DNA, mixing and reacting to obtain a reaction solution, testing the fluorescence intensity of the reaction solution by a fluorescence spectrometry, and drawing a standard curve according to the relationship between the concentration and the fluorescence intensity;(2) adding a to-be-detected sample solution and the reducing agent into the mixed solution of graphdiyne and the group-modified double-stranded DNA, mixing and reacting to obtain the reaction solution, testing the fluorescence intensity of the reaction solution according to fluorescence spectrometry, and quantifying the concentration of the copper ions in the to-be-detected sample according to thestandard curve obtained in the step (1). By selecting the graphite alkyne in a detection system, the situation that the alkynyl groups are used for modification is avoided, the detection efficiency can be greatly improved, and the operation steps are simplified.

Description

technical field [0001] The invention belongs to the field of detection, and relates to a detection method, kit and application of copper ions based on graphyne and click chemistry. Background technique [0002] Cu 2+ It is an essential element necessary for the ligands of a large number of oxidoreductases and metalloproteins in the metabolic process of human life. These enzymes and proteins, such as tyrosinase, cytochrome c oxidase, and ceruloplasmin, play important roles in electron transfer, oxidation, and reduction. Therefore, Cu in biological systems 2+ content should be strictly controlled. Unusual levels of free Cu 2+ May act as a catalyst for the generation of oxygen and protein free radicals, leading to severe neurological diseases such as Alzheimer's, Parkinson's and Wilson's diseases. Early diagnosis is of great significance to control the occurrence and development of these diseases. For example, Cu in urine 2+ The high level is one of the clinical and diag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/6486G01N2021/6417G01N2021/6432
Inventor 鲍世韵王斗葛晨晨刘利平刘权吴宗泽
Owner SHENZHEN PEOPLES HOSPITAL
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