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Method for quantitatively detecting glutathione by using labeling-free copper-indium-sulfur fluorescent probe synchronous fluorescence method

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

Inactive Publication Date: 2018-02-09
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 quantitatively detecting glutathione by using labeling-free copper-indium-sulfur fluorescent probe synchronous fluorescence method
  • Method for quantitatively detecting glutathione by using labeling-free copper-indium-sulfur fluorescent probe synchronous fluorescence method
  • Method for quantitatively detecting glutathione by using labeling-free copper-indium-sulfur fluorescent probe synchronous fluorescence method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: 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. After being fully dissolved, 157 μL of 3-mercaptopropionic acid was added dropwise to the system with a pipette. At this time, the solution changed from a colorless, clear and transparent state to a light yellow precipitate. Then use 2mol / L NaOH solution to adjust pH=11.3. In the process of adjusting the pH, it is obvious that the precipitation gradually disappears until it returns to a colorless and clear solution state. After the magnet is stirred for 10 minutes, add 0.0228 g of thiourea and continue stirring until it is completely dissolved. Transfer the synthetic solution to a polytetrafluoroethylene reactor and place it in an oven at 150°C for 21 hours and allow it to cool to room temperature naturally. A light yellow clear solution can be obtained, indicating that the copper indiu...

Embodiment 2

[0032] Example 2: Solution configuration

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

[0034] (2) Configuration of 0.2mol / L, 25℃, 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 is dissolved in 10mL high purity water; ②0.2mol / L NaH 2 PO 4 Solution: weigh 0.312g NaH 2 PO 4 ·2H 2 O is dissolved in 10mL high-purity water; ③take 1.9mL NaH 2 PO 4 Solution and 8.1mL Na 2 HPO 4 Solution, dilute to 20mL, store in the refrigerator for later use;

[0036] (3) The configuration of different concentrations of copper chloride solution:

[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 25μL, 50μL, 75μL, 100μL, 125μL, 150μL, etc. (1M) CuCl 2 The solution is...

Embodiment 3

[0040] Example 3: The synchronous fluorescence method is used as the detection method, and the copper indium sulfur quantum dots are used for specific detection of glutathione to determine the detection time (using a fluorometer for Agilent)

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

[0042] (2) Add 180μL of copper indium sulfur quantum dot mother liquor to the centrifuge tube, 200μL (0.02mol / L) PBS solution and mix well, then add 40μL (1mM) copper ion solution as the fluorescence quencher, shake well Let it sta...

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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 sulphur optical probe combined with a synchronous fluorescence method for optical detection and its application to the quantitative detection of biological thiol 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, it has a near-infrared fluorescent property when its particle size is about 2-4nm. Compared with traditional quantum dots, the new type of copper indium sulfur quantum dots do not contain toxic metal elements such as mercury and cadmium, have good light stability, and emit light in the near-infrared region (about 660nm), making them more suitable for biological detection. However, in recent years, people have widely used copper indium sulfur as a thin film ...

Claims

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Application Information

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Patent Type & Authority Applications(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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