A detection method, kit and application of copper ions based on graphdiyne and click chemistry

A detection method and click chemistry technology, applied in the detection field, can solve the problems of reducing the repeatability of the reaction and increasing the complexity of the reaction, and achieve the effects of excellent fluorescence resonance energy transfer capability, low detection limit and high detection sensitivity.

Active Publication Date: 2022-06-17
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

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  • A detection method, kit and application of copper ions based on graphdiyne and click chemistry
  • A detection method, kit and application of copper ions based on graphdiyne and click chemistry
  • A detection method, kit and application of copper ions based on graphdiyne and click chemistry

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Embodiment 1

[0058] The present embodiment provides a method for detecting copper ions based on graphdiyne and click chemistry, comprising the following steps:

[0059] (1) 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 were added to the PBS buffer, and mixed at 95°C for 5 minutes to obtain a group-modified double-stranded DNA, which was stored at 4°C;

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

[0061] (3) To the mixture of graphdiyne and group-modified double-stranded DNA obtained in step (2), add 20 μL of copper ion standard solution and 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 was 10 μg / mL, and the concentration of the group-modified doubl...

Embodiment 2

[0079] The present embodiment provides a method for detecting copper ions based on graphdiyne and click chemistry, comprising the following steps:

[0080] (1) 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 were added to the PBS buffer, and mixed at 80°C for 8 minutes to obtain a group-modified double-stranded DNA, which was stored at 4°C;

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

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

Embodiment 3

[0092] The present embodiment provides a method for detecting copper ions based on graphdiyne and click chemistry, comprising the following steps:

[0093] (1) 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 were added to the PBS buffer, and mixed at 100°C for 3 minutes to obtain a group-modified double-stranded DNA, which was stored at 4°C;

[0094] (2) Mixing the group-modified double-stranded DNA obtained in step (1) and 7.5 μg of graphdiyne in 0.5 mL of PBS buffer by vortexing for 10 s to obtain a mixture of graphyne and group-modified double-stranded DNA ;

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

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Abstract

The present invention provides a kind of detection method, test kit and application of copper ion based on graphyne and click chemistry; Said detection method comprises the following steps: (1) in the mixed solution of graphyne and the double-stranded DNA of group modification Add the copper ion standard solution and reducing agent of concentration gradient, mix, react, obtain reaction liquid, then test the fluorescence intensity of reaction liquid by fluorescence spectrometry, draw standard curve according to the relation of concentration and fluorescence intensity; (2) graphyne Add the sample solution to be tested and the reducing agent to the mixed solution of the double-stranded DNA modified by the group, mix and react to obtain the reaction solution, and then test the fluorescence intensity of the reaction solution according to the fluorescence spectrometry, according to the standard curve obtained in step (1) Quantify the concentration of copper ions in the sample to be tested; the use of graphyne in the detection system can avoid modification with alkynyl groups, greatly improve the detection efficiency, and simplify the operation steps.

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 graphdiyne 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 process of human life metabolism. 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. Abnormal 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 disease, Parkinson's disease and Wilson's disease. Early diagnosis is of great significance to control the occurrence and development of these diseases. For example, Cu in urine 2+ The high level of Wilson di...

Claims

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Application Information

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Patent Type & Authority Patents(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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