Method for quantitatively detecting alanine transaminase in solution based on copper nanocluster fluorescent probe

An alanine aminotransferase and copper nano-cluster technology, which is applied in fluorescence/phosphorescence, measuring devices, material analysis by optical means, etc., can solve the problems of low detection sensitivity, inaccuracy and high detection limit of detection purity, and achieves a synthesis method. Simple, good fluorescence performance, low detection limit effect

Inactive Publication Date: 2019-04-09
TIANJIN NORMAL UNIVERSITY
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  • Claims
  • Application Information

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Problems solved by technology

However, this result may be inaccurate, and these methods usually have high detection limits or require expensive reagents such as enzymes or coenzyme factors. Therefore, in order to solve these problems, many new methods for detecting the content of alanine aminotransferase have been developed. Established, such as: colorimetry, chromatography, etc., but due to the harsh conditions of the detection pre-treatment, high requirements for detection purity, low detection sensitivity and other issues have limited the wide application of these methods

Method used

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  • Method for quantitatively detecting alanine transaminase in solution based on copper nanocluster fluorescent probe
  • Method for quantitatively detecting alanine transaminase in solution based on copper nanocluster fluorescent probe
  • Method for quantitatively detecting alanine transaminase in solution based on copper nanocluster fluorescent probe

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Experimental program
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Effect test

Embodiment 1

[0025] The preparation of copper nanoclusters with glutathione as a stabilizer is carried out according to the following steps at room temperature:

[0026] (1) Preparation of 0.1 M copper chloride solution: weigh 1.7048 g CuCl 2 ∙2H 2 O was dissolved in 100 mL of high-purity water, fully dissolved for later use;

[0027] (2) Preparation of copper nanoclusters: at room temperature, 0.28 g glutathione was weighed and dissolved in 15 mL H 2 O, to which was added 450 mL CuCl 2 (0.1M), after fully reacting, add 0.1 g ascorbic acid (AA), then add 1 mL NaOH (1M), react for 1 h, until the white suspension is completely dissolved and turns into a light yellow clear solution, which proves that the copper nanoclusters form. By transmission electron microscopy (TEM) ( figure 1 ) It can be seen that the copper nanoclusters are uniformly dispersed and the particle size is small.

Embodiment 2

[0029] Copper nano-clusters are used as a method for fluorescent probe specific detection of alanine aminotransferase, which is characterized in that it is carried out according to the following steps:

[0030] (1) Preparation of tiopronin mother liquor: weigh 0.1000 g tiopronin and dissolve it in 5 mL of high-purity water; dilute the mother liquor to a low concentration of 1 mg / mL, and store it at low temperature for later use;

[0031] (2) Preparation of a series of different concentration solutions of alanine aminotransferase:

[0032] Prepare alanine aminotransferase solutions with concentrations of 1, 5, 50, 100, 500, and 1000 U / L respectively, and store them at low temperature for later use;

[0033] (3) Evenly disperse the prepared copper nanoclusters based on glutathione as a stabilizer in high-purity water, prepare a detection system with a concentration of 0.675 mM and a volume of 4 mL, and measure the concentration at this time using a fluorescence spectrophotometer...

Embodiment 3

[0039] 1. Refer to Example 1 for the preparation of copper nanoclusters using glutathione as a protective agent;

[0040] 2. Determination of excitation and emission spectra of copper nanoclusters stabilized with glutathione:

[0041] Disperse copper nanoclusters in high-purity water to measure the fluorescence excitation spectrum and fluorescence emission spectrum of the material, such as figure 2 As shown, the maximum excitation wavelength of copper nanoclusters is 354 nm, and under the excitation of the maximum excitation wavelength, the fluorescence emission wavelength is 632 nm.

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Abstract

The invention discloses a method for quantitatively detecting alanine transaminase in a solution based on a copper nanocluster fluorescent probe, wherein a copper nanocluster with glutathione as a protective agent is used as a fluorescent probe, and the content of alanine transaminase in the solution is specifically detected through a fluorescence "off-on" mode. The label-free, highly sensitive and selective detection of alanine transaminase is achieved by adopting the fluorescence "off-on" mode. The method is simple and rapid, the specific detection of alanine transaminase can be realized, the detection linear range is wide, and the detection limit is low. The method in the invention has good detection sensitivity and selectivity and has good application prospects in disease detection, clinical application and other aspects.

Description

[0001] This patent has been supported by the National Natural Science Foundation of China General Project 21375095, Tianjin Natural Science Foundation Youth Project (No.17JCQNJC05800), Tianjin Normal University Doctoral Fund (No.52XB1510), Tianjin Normal University "Inorganic-Organic Hybrid Functional Materials Chemistry Education Department" Key Laboratory", "Tianjin Key Laboratory of Functional Molecular Structure and Performance" open fund project and Tianjin Normal University "Future Thousand Talents Program" project (WLQR201711, WLQR201814). technical field [0002] The invention belongs to the application field of metallic copper nanoclusters in fluorescence sensing, and specifically relates to a label-free, efficient and specific detection of alanine aminotransferase in a complex system using a fluorescent "off-on" mode using copper nanoclusters as a fluorescent probe content method. Background technique [0003] Alanine aminotransferase (ALT), also known as glutamate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432
Inventor 李妍吴晓曼张菲冯子硕来萌萌
Owner TIANJIN NORMAL UNIVERSITY
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