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Metallothionein detection method based on carbon quantum dot-gold nanoparticle system

A technology of metallothionein and gold nanoparticles, which is applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of lack of selectivity and sensitivity, difficult to popularize methods, cumbersome procedures, etc., and achieve good photostability and water solubility Sexuality, good effect, and wide excitation spectrum

Active Publication Date: 2020-08-21
TAIYUAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of these methods lack sufficient selectivity and sensitivity, and due to the need for expensive and complex equipment and cumbersome procedures, these methods are difficult to popularize.

Method used

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  • Metallothionein detection method based on carbon quantum dot-gold nanoparticle system
  • Metallothionein detection method based on carbon quantum dot-gold nanoparticle system
  • Metallothionein detection method based on carbon quantum dot-gold nanoparticle system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Weigh 0.5g L-glutamic acid and 1.04g 4-aminosalicylic acid respectively, add to 20mL deionized water, stir and dissolve to obtain a mixed solution.

[0037] The mixed solution was transferred to a 50 mL stainless steel autoclave lined with polytetrafluoroethylene, heated to 200 ° C for 7 h. After the reaction was completed, it was naturally cooled to room temperature to obtain the original solution of carbon quantum dots.

[0038] Centrifuge the original solution of carbon quantum dots at 10000r / min for 15min, take the supernatant, and filter it with a 0.22µm microporous filter to obtain a clear orange solution, place it in a dialysis membrane (MWCO=1kDa), and dialyze it in deionized water 24h.

[0039] The carbon quantum dot solution after dialysis is freeze-dried to prepare black nitrogen-doped carbon quantum dot solid powder.

[0040] It has been determined that the optimal excitation wavelength of the aqueous solution of the prepared carbon quantum dots is 390nm. ...

Embodiment 2

[0042] Take the carbon quantum dot solid powder prepared in Example 1, disperse it in deionized water, and prepare a carbon quantum dot solution with a concentration of 2.0 mg / mL.

[0043]Measure 0µL, 12.5µL, 25µL, 37.5µL, 50µL, 62.5µL, 75µL, 87.5µL, 100µL, 112.5µL, 125µL, 137.5µL, 150µL of gold nanoparticle solution with a concentration of 6.96nM, and add 5µL with a concentration of 2.0mg / mL carbon quantum dot solution, react at room temperature for 5min, dilute to 500µL with ultrapure water, and prepare gold nanoparticles with concentrations of 0, 0.174, 0.348, 0.522, 0.696, 0.87, 1.044, 1.218, 1.392, 1.566, 1.74, A series of carbon quantum dots-gold nanoparticles standard solutions of 1.914 and 2.088nM.

[0044] A fluorescence spectrophotometer was used to detect the fluorescence intensity of the above-mentioned series of carbon quantum dots-gold nanoparticles standard solutions at an excitation wavelength of 390 nm. Test results such as figure 1 , where the concentratio...

Embodiment 3

[0046] Weigh a small amount of metallothionein freeze-dried powder, dissolve it in 10mL sterilized water, and prepare the original solution of metallothionein, and use the Ellman's reagent quantitative method to determine its concentration as 1.17×10 -6 mol / L.

[0047] Measure 5 μL of carbon quantum dot solution with a concentration of 2.0 mg / mL and 150 μL of gold nanoparticle solution with a concentration of 6.96 nM in different microcentrifuge tubes, and react at room temperature for 5 minutes. Then add 0µL, 5µL, 10µL, 20µL, 30µL, 40µL, 50µL, 60µL, 70µL, 80µL, 90µL metallothionein solution respectively, and add sodium citrate-hydrochloric acid buffer solution (pH=3.0 )30μL, and the volume was adjusted to 500μL with deionized water, in which the concentration of metallothionein ( c MTs ) are 0, 1.2, 2.3, 4.7, 7.0, 9.4, 12, 14, 16, 19, 21×10 -8 mol / L, heated at 50°C for 30 minutes to fully react, and a series of metallothionein standard solutions were obtained.

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Abstract

The invention discloses a metallothionein detection method based on a carbon quantum dot-gold nanoparticle system. The preparation method comprises the following steps of: carrying out hydrothermal reaction on L-glutamic acid and 4-aminosalicylic acid to prepare green fluorescent carbon quantum dots; respectively detecting the fluorescence intensity F0 of a carbon quantum dot-gold nanoparticle standard solution and the fluorescence intensity F of a series metallothionein standard solution, establishing a linear relation between (F-F0) / F0 and the metallothionein concentration cMTs, detecting the fluorescence intensity of a to-be-detected sample solution, and substituting the fluorescence intensity into the linear relation to calculate the metallothionein concentration of the to-be-detectedsample solution. The method disclosed by the invention can be used for simply, conveniently, quickly and sensitively detecting metallothionein, and is suitable for detecting metallothionein in any aqueous solution system, especially urine.

Description

technical field [0001] The invention relates to a method for detecting metallothionein, in particular to a method for quantitatively detecting the concentration of metallothionein by using a carbon quantum dot-gold nanoparticle system. Background technique [0002] Metallothionein (MTs) is a low molecular weight (Mr<10kDa), cysteine-rich metal-binding protein, which widely exists in various tissues and organs of various animals, higher plants, microorganisms and humans. Because of its easy combination with heavy metals, it plays an important role in heavy metal detoxification, scavenging free radicals, regulating intracellular balance and maintaining the steady state of metal ion concentration. In addition, MTs are also related to physiological processes such as growth and development of the body and certain diseases. [0003] Based on the responsiveness of MTs to heavy metal pollution and the statistical correlation between the content of MTs in aquatic animals and the ...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/68
CPCG01N21/6428G01N21/6402G01N33/68G01N2021/6432G01N2333/825
Inventor 段炼孙玥赵慧君杜晓宇
Owner TAIYUAN UNIV OF TECH
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