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Preparation method of nanogold cluster and sensitization detection method of nanogold cluster in small biological molecules

A gold nano-cluster and gold nano-cluster technology, applied in the field of nano-gold clusters, can solve the problems of expensive enzyme, inconvenient storage, complicated electrochemical method operation, etc., and achieve the effect of sensitizing detection of uric acid content

Pending Publication Date: 2022-04-12
中检国研长春科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chromatography requires expensive instruments, and electrochemical methods are more complicated to operate. Enzymatic methods mainly use two kinds of enzymes (uric acid oxidase and peroxidase) and color source substances. Enzymes are expensive and inconvenient to store. It is greatly affected by external conditions (such as temperature, pH value, especially organic solvents), which limit the application of these methods

Method used

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  • Preparation method of nanogold cluster and sensitization detection method of nanogold cluster in small biological molecules
  • Preparation method of nanogold cluster and sensitization detection method of nanogold cluster in small biological molecules
  • Preparation method of nanogold cluster and sensitization detection method of nanogold cluster in small biological molecules

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Preparation of gold nanoclusters

[0021] Prepare 100mmol / L HAuCl4 and 2mol / L NaOH solution. Add 200mg of BSA to 10mL of ultrapure water, stir in a water bath at 40°C until dissolved, quickly add 10mL of 100mmol / L HAuCl4 aqueous solution to the BSA solution under vigorous stirring, mix for 10min, then add 0.5mL of 2mol / L NaOH solution , stirred gently at 37°C for 40h.

[0022] Nanocluster Separation and Purification

[0023] The prepared nano-gold clusters were stored in the dark for 48 hours, and then poured into a 20,000MWCO dialysis bag for dialysis for 48 hours (the water was changed every 8 hours). After dialysis, the mixed solution was filtered through a 0.22 μm microporous membrane with a syringe to remove excess impurities with large particle sizes. The finally obtained solution was stored at 4°C in the dark.

[0024] Experimental Materials

[0025] sample:

[0026] The BSA-AuNCs produced in Example 1 above. Blood samples were taken from 3 healthy adults....

Embodiment 2

[0033] Preparation of gold nanoclusters

[0034] Prepare 1mmol / L HAuCl4 and 0.5mol / L NaOH solution. Add 10mg of GSH into 1mL of ultrapure water, stir in a water bath at 25°C until dissolved, quickly add 1mL of 1mmol / L HAuCl4 aqueous solution to the GSH solution under vigorous stirring and mix for 3 minutes, then add 0.5mol / L of NaOH solution 0.1 mL, stirred gently at 37°C for 10h.

[0035] Nanocluster Separation and Purification

[0036] The prepared nano-gold clusters were stored in the dark for 12 hours, and then poured into a 20000MWCO dialysis bag for dialysis for 12 hours (the water was changed every 4 hours). After dialysis, the mixed solution was filtered through a 0.22 μm microporous membrane with a syringe to remove excess impurities with large particle sizes. The finally obtained solution was stored at 0°C in the dark.

[0037] The GSH-AuNCs produced in Example 1 above. Blood samples were taken from 3 healthy adults.

[0038] experimental method:

[0039] Esta...

Embodiment 3

[0044] Preparation of gold nanoclusters

[0045] Prepare 10mmol / L HAuCl4 and 1mol / L NaOH solution. Add 125mg CEW to 2.5mL ultrapure water, stir in a water bath at 37°C until dissolved, quickly add 2.5mL 10mmol / L HAuCl4 aqueous solution to the CEW solution under vigorous stirring, mix for 5 minutes, then add 1mol / L NaOH solution 0.25mL, stirred gently at 37°C for 24h.

[0046] Nanocluster Separation and Purification

[0047] The prepared nano-gold clusters were stored in the dark for 24 hours, and then poured into a 20,000MWCO dialysis bag for dialysis for 24 hours (the water was changed every 8 hours). After dialysis, the mixed solution was filtered through a 0.22 μm microporous membrane with a syringe to remove excess impurities with large particle sizes. The finally obtained solution was stored at 4°C in the dark.

[0048] Experimental Materials

[0049] CEW-AuNCs produced in Examples 1, 2, and 3 above. Blood samples were taken from 3 healthy adults.

[0050] experime...

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Abstract

The invention discloses a nanogold cluster preparation method and a biological small molecule sensitization detection method, uric acid generates hydrogen peroxide under the catalysis of urate oxidase, a substrate 3, 3 ', 5, 5'-tetramethyl benzidine is oxidized into ox-TMB by using peroxidase activity of the synthesized gold nanocluster and a catalytic product H2O2, at the moment, an emission spectrum of AuNCs is overlapped with an absorption spectrum of ox-TMB, and the concentration of the ox-TMB is determined. According to the method disclosed by the invention, fluorescence resonance energy transfer occurs between the strongly fluorescent AuNCs and ox-TMB, a BSA-AuNCs fluorescence signal achieves a quenching effect, and the method not only solves the problem that use conditions of a traditional enzyme method are restricted, but also reduces a detection line and realizes sensibilization detection of uric acid molecules. Due to the fact that the emission spectrum of the AuNCs is overlapped with the absorption spectrum of the ox-TMB, fluorescence resonance energy transfer occurs between the AuNCs with strong fluorescence and the ox-TMB, the fluorescence signal of the AuNCs is obviously reduced, and the purpose of sensitizing and detecting the content of the uric acid is achieved.

Description

technical field [0001] The invention belongs to the technical field of nano-gold clusters, and in particular relates to a preparation method of nano-gold clusters capable of detecting uric acid content in blood. Background technique [0002] In recent decades, gold nanoclusters (AuNCs), as a new type of non-toxic and harmless luminescent nanomaterial, have been widely used in biomedical research, and advances in nanoscale technology contribute to the diagnosis, treatment, and prevention of diseases . [UpadhyayY, Kumar R, Kumar S A, et al.Vitamin B6 cofactors conjugated ovalbumin-stabilized gold nanoclusters:Application in alkaline phosphatase activity detection and generating white-light emission[J].Microchemical Journal,2020,156:104859.]AuNCs is a A new type of ultra-small fluorescent nanomaterials, [Yang Qunfeng, Liu Jianyun, Chen Huaping, et al. Preparation of noble metal nanoclusters and their application in biological detection [J]. Advances in Chemistry, 2011,23(5):88...

Claims

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Application Information

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IPC IPC(8): G01N21/64C09K11/58B82Y20/00B82Y40/00
Inventor 邹骏李秀勇丛剑涵
Owner 中检国研长春科技有限公司
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