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Heparin detection method for copper nano-cluster based on denatured bovine serum protein as template

A technology of bovine serum albumin and copper nanoclusters, which is applied in the field of aqueous phase synthesis of copper nanoclusters, can solve the problems of safety and biocompatibility, biological detection limits, etc., and achieve wide detection linear range and detection limit The effect of low and uniform particle size

Inactive Publication Date: 2018-12-11
TIANJIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional nanomaterials such as quantum dots and metal nanoclusters have been well used as fluorescent probes. However, the presence of heavy metals in quantum dots makes their safety and biocompatibility still questionable, which makes them suitable for biological detection. Restricted

Method used

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  • Heparin detection method for copper nano-cluster based on denatured bovine serum protein as template
  • Heparin detection method for copper nano-cluster based on denatured bovine serum protein as template
  • Heparin detection method for copper nano-cluster based on denatured bovine serum protein as template

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Experimental program
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Effect test

Embodiment 1

[0039] 0.1M CuCl 2 Solution preparation: weigh 1.7050g CuCl 2 Dissolve in an appropriate amount of high-purity water, transfer to a 100mL volumetric flask to constant volume, and label for use;

[0040] Denaturation of bovine serum albumin: weigh 0.1200g of bovine serum albumin and dissolve it in 30mL of high-purity water, fully dissolve under stirring conditions, weigh 0.0040g of NaBH 4 Add to the above bovine serum albumin solution, fully react for 1h, heat the above mixed solution at 70°C for 30min, and cool to room temperature;

[0041] Synthesis of copper nanoclusters: Add the prepared CuCl to the denatured bovine serum albumin solution under agitation 2 250 μL of the solution. After fully reacting, add 0.7 mL of hydrazine hydrate to the above mixed solution, and react for 2 hours under stirring until the solution turns light yellow;

Embodiment 2

[0043] 0.1M CuCl 2 Solution preparation: weigh 1.7050g CuCl 2 Dissolve in an appropriate amount of high-purity water, transfer to a 100mL volumetric flask to constant volume, and label for use;

[0044] Denaturation of bovine serum albumin: weigh 0.1200g of bovine serum albumin and dissolve it in 30mL of high-purity water, fully dissolve under stirring conditions, weigh 0.0040g of NaBH 4 Add to the above bovine serum albumin solution, fully react for 1h, heat the above mixed solution at 70°C for 30min, and cool to room temperature;

[0045] Synthesis of copper nanoclusters: Add the prepared CuCl to the denatured bovine serum albumin solution under agitation 2 250 μL of the solution. After fully reacting, add 0.7 mL of hydrazine hydrate to the above mixed solution, and react for 6 hours under stirring until the solution turns light yellow;

Embodiment 3

[0047] 0.1M CuCl 2 Solution preparation: weigh 1.7050g CuCl 2 Dissolve in an appropriate amount of high-purity water, transfer to a 100mL volumetric flask to constant volume, and label for use;

[0048] Denaturation of bovine serum albumin: weigh 0.0800g of bovine serum albumin and dissolve it in 30mL of high-purity water, fully dissolve under stirring conditions, weigh 0.0040g of NaBH 4 Add to the above bovine serum albumin solution, fully react for 1h, heat the above mixed solution at 70°C for 30min, and cool to room temperature;

[0049] Synthesis of copper nanoclusters: Add the prepared CuCl to the denatured bovine serum albumin solution under agitation 2 250 μL of the solution. After fully reacting, add 0.7 mL of hydrazine hydrate to the above mixed solution, and react for 6 hours under stirring until the solution turns light yellow;

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Abstract

The invention discloses a heparin detection method for a copper nano-cluster based on a denatured bovine serum protein as template. The method comprises the following steps: forming a heparin detection system with a phosphoric acid buffer aqueous solution, a copper nano-cluster dispersion system and a to-be-tested sample; detecting fluorescence intensity change before and after adding the to-be-tested sample; and comparing with a standard curve to obtain the content of heparin in the to-be-tested sample. The method synthesizes the copper nano-cluster by taking the denatured bovine serum protein as the template by means of a one-pot method. The synthesized material is uniform in particle size, good in fluorescence property and stable in property, and the light-emitting position is near 642.04 nm and belongs to a near infrared area, so that interference of autofluorescence is avoided. The method can detect the heparin content directly and quickly, is wide in detection linear range and low in detection limit and has a very good application prospect in the fields of biological system detection, clinical applications and the like.

Description

technical field [0001] The invention belongs to the field of preparation of metal nanoclusters and its application in fluorescence sensing, in particular to an aqueous phase synthesis method of copper nanoclusters based on denatured bovine serum albumin as a template and the use of fluorescence quenching "turn off" Application of the model in label-free, efficient and selective detection of heparin. Background technique [0002] Heparin is an acidic mucopolysaccharide, which widely exists in the liver, kidney, thymus, and blood of mammals. It has unique properties in physiology and food analysis, and has important applications in cardiovascular diseases and the treatment of coagulation, such as the prevention and treatment of thromboembolism Diseases, disseminated intravascular coagulation, and anticoagulation in vitro. In cardiovascular surgery or to avoid thrombosis, it is crucial to monitor the level of heparin, because excess heparin can cause some negative effects, suc...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432
Inventor 李妍吴晓曼李欣格张菲
Owner TIANJIN NORMAL UNIVERSITY
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