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Method for quantitative detection of glutathione by simultaneous fluorescence method with label-free copper indium sulfur fluorescent probe

A technology of glutathione and fluorescent probe, applied in the field of quantitative detection of biothiol glutathione, can solve the problems of incompleteness, low accuracy and sensitivity, poor symmetry, etc., and achieve simplified synthesis steps, improved sensitivity, Solve the effect of incomplete peak shape

Inactive Publication Date: 2020-01-07
TIANJIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in recent years, copper indium sulfide has been widely used in thin film materials of solar cells, but it is rarely used as a fluorescent probe in detection
The main reason is that when copper indium sulfur is used as a fluorescent probe for quantitative detection using traditional fluorescence methods, it shows a relatively incomplete and poorly symmetrical fluorescence peak shape, resulting in low accuracy and sensitivity

Method used

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  • Method for quantitative detection of glutathione by simultaneous fluorescence method with label-free copper indium sulfur fluorescent probe
  • Method for quantitative detection of glutathione by simultaneous fluorescence method with label-free copper indium sulfur fluorescent probe
  • Method for quantitative detection of glutathione by simultaneous fluorescence method with label-free copper indium sulfur fluorescent probe

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: the synthesis of copper indium sulfur quantum dots

[0029] Weigh 0.0256g CuCl respectively 2 2H 2 O and 0.0573g In(NO 3 ) 3 ·5H 2 O was dissolved in 10.5 mL of high-purity water, and after it was fully dissolved, pipette 157 μL of 3-mercaptopropionic acid into the system dropwise. At this time, the solution changed from a colorless, clear and transparent state to a pale yellow precipitate. Then adjust the pH to 11.3 with 2 mol / L NaOH solution. In the process of adjusting the pH, it can be clearly found that the precipitate gradually disappears until it returns to a colorless and clear solution state. After being stirred by the magnet for 10 minutes, 0.0228 g of thiourea was added and the stirring was continued until it was completely dissolved. Transfer the synthesis solution to a polytetrafluoroethylene reactor, heat it in an oven at 150°C for 21 hours, and let it cool down to room temperature naturally. A light yellow clear solution can be obtain...

Embodiment 2

[0032] Embodiment 2: the configuration of solution

[0033] (1) Preparation of copper indium sulfur quantum dot mother liquor: accurately weigh 0.0190 g of purified copper indium sulfur quantum dot powder, dissolve in 10 mL of high-purity water, and shake well for later use;

[0034] (2) Configuration of 0.2mol / L, 25°C, pH=7.4 PBS buffer solution:

[0035] ①0.2mol / L Na 2 HPO 4 Solution: weigh 0.716g Na 2 HPO 4 12H 2 O dissolved in 10mL high-purity water; ②0.2mol / L NaH 2 PO 4 Solution: weigh 0.312g NaH 2 PO 4 2H 2 O was dissolved in 10mL high-purity water; ③Take 1.9mLNaH 2 PO 4 solution and 8.1mL Na 2 HPO 4 solution, dilute to 20mL, and store in the refrigerator for later use;

[0036] (3) Configuration of copper chloride solutions with different concentrations:

[0037] Accurately weigh 0.8541g of copper chloride and dissolve it in 5mL of high-purity water to prepare CuCl with a concentration of 1M 2 solution; respectively take 25 μL, 50 μL, 75 μL, 100 μL, 125 ...

Embodiment 3

[0040] Embodiment 3: Using the synchronous fluorescence method as the detection method, copper indium sulfur quantum dots are used to specifically detect glutathione to determine the detection time (Using a fluorescence photometer is Agilent, USA)

[0041] (1) Add 180 μL of copper indium sulfur quantum dot mother solution and 200 μL (0.02mol / L) PBS solution to the centrifuge tube and mix well, then add 40 μL of a series of gradient concentrations (0.1mM-1M) copper ion solution (chloride Aqueous copper chloride solution) was used as a fluorescence quencher, and high-purity water was added to make the volume to 4 mL. After shaking well and standing for different times, the fluorescence spectrophotometer was adjusted to the detection mode of the synchronous fluorescence method, and the displacement difference was set to 60nm for detection.

[0042] (2) Add 180 μL copper indium sulfur quantum dot mother solution and 200 μL (0.02mol / L) PBS solution to the centrifuge tube in turn, m...

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Abstract

The invention discloses a method for quantitatively detecting glutathione by using a labeling-free copper-indium-sulfur fluorescent probe synchronous fluorescence method. According to the present invention, copper-indium-sulfur is used as a fluorescent probe, a synchronous fluorescence method is used as a detection method, the detection disadvantage of copper-indium-sulfur quantum dots is effectively used as the breakthrough point, and the specific detection of glutathione is achieved by using the copper-indium-sulfur quantum dots as the fluorescent probe; the used copper-indium-sulfur quantumdots are labelling-free and unmodified, such that the synthesis steps are greatly simplified; the detection system has the significantly-improved sensitivity and significantly-improved selectivity; and the method has advantages of complete and symmetrical peak shape, high accuracy and high sensitivity.

Description

technical field [0001] The invention belongs to the technical field of biological analysis and detection, and mainly relates to a label-free copper indium sulfur optical probe combined with a synchronous fluorescence method for optical detection and its application to the quantitative detection of biothiol glutathione. Background technique [0002] At present, the ternary semiconductor material copper indium sulfur has been widely used in the production of thin film materials for solar cells. According to experiments, when its particle size is about 2-4nm, it will have near-infrared fluorescence properties. Compared with traditional quantum dots, the new copper indium sulfur quantum dots do not contain toxic metal elements such as mercury and cadmium, have good photostability, and the emission wavelength is in the near-infrared region (about 660nm), which is more suitable for biological detection. However, copper indium sulfide has been widely used in thin film materials of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 李妍王璐孙玉绣张菲
Owner TIANJIN NORMAL UNIVERSITY
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