A Colorimetric Sensing Method for Hydrogen Peroxide Based on Fluorescent Polymer Mimic Enzymes

A fluorescent polymer, hydrogen peroxide technology, applied in fluorescence/phosphorescence, analysis by chemical reaction of materials, color/spectral property measurement, etc. complex problems, to achieve the effect of long service life, simple method and optimized catalytic activity

Active Publication Date: 2020-05-01
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the preparation process of these simulated enzyme materials is complicated, the cost is high, and the synthesis conditions are not easy to control, which limits their further wide application and promotion.

Method used

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  • A Colorimetric Sensing Method for Hydrogen Peroxide Based on Fluorescent Polymer Mimic Enzymes
  • A Colorimetric Sensing Method for Hydrogen Peroxide Based on Fluorescent Polymer Mimic Enzymes
  • A Colorimetric Sensing Method for Hydrogen Peroxide Based on Fluorescent Polymer Mimic Enzymes

Examples

Experimental program
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Effect test

Embodiment 1

[0027] (1) Add 0.1mL of concentrated sulfuric acid (98% by mass) dropwise to 5mL of diethylenetriamine, and stand still for 20 minutes in an ice-water bath. After the reaction, a milky white fluorescent polymer mimic enzyme can be obtained without any treatment. ;

[0028] (2) Dispersing the fluorescent polymer-mimicking enzyme obtained in step (1) into water to prepare a fluorescent polymer-mimicking enzyme dispersion with a concentration of 1 mg / mL;

[0029] (3) Add 10 μL of the fluorescent polymer mimic enzyme dispersion prepared in step (2) to 500 μL of NaAc-HAc buffer solution with pH = 3.5; then add 100 μL of 3,3’ at a concentration of 1 mM to the reaction system , 5,5'-tetramethylbenzidine (TMB) ethanol solution was used as the colorimetric reaction substrate; then add 3.25 μL, 6.5 μL, 9.75 μL, 13 μL, 16.25 μL, 26 μL, 32.5 μL of peroxide with a concentration of 0.01M Hydrogen solution, incubate at 35°C for 15 minutes to obtain hydrogen peroxide solutions with hydrogen ...

Embodiment 2

[0034](1) Add 0.5mL of concentrated sulfuric acid (98% by mass) dropwise to 4mL of diethylenetriamine, and stand still for 20 minutes in an ice-water bath. After the reaction, a milky white fluorescent polymer mimic enzyme can be obtained without any treatment. ;

[0035] (2) Dispersing the fluorescent polymer-mimicking enzyme obtained in step (1) into water to prepare a fluorescent polymer-mimicking enzyme dispersion with a concentration of 2.5 mg / mL;

[0036] (3) Add 25 μL of the fluorescent polymer mimic enzyme dispersion prepared in step (2) to 600 μL of NaAc-HAc buffer solution with pH = 3.5; then add 100 μL of 3,3’ at a concentration of 2 mM to the reaction system , 5,5'-tetramethylbenzidine (TMB) ethanol solution was used as the colorimetric reaction substrate; then 3.63 μL, 6.5 μL, 10.87 μL, 14.5 μL, 18.13 μL, 29 μL, 36.25 μL of 0.01 M Hydrogen peroxide solution, incubate at 35°C for 15 minutes to obtain hydrogen peroxide solutions with hydrogen peroxide concentration...

Embodiment 3

[0041] (1) Add 1mL of concentrated sulfuric acid (98% by mass) dropwise to 3mL of diethylenetriamine, and stand still for 30 minutes in an ice-water bath. After the reaction, a milky white fluorescent polymer mimetic enzyme can be obtained without any treatment;

[0042] (2) Dispersing the fluorescent polymer-mimicking enzyme obtained in step (1) into water to prepare a fluorescent polymer-mimicking enzyme dispersion with a concentration of 5 mg / mL;

[0043] (3) Add 50 μL of the fluorescent polymer mimic enzyme dispersion prepared in step (2) to 700 μL of NaAc-HAc buffer solution with pH=4.0; then add 200 μL of 3,3’ , 5,5'-Tetramethylbenzidine (TMB) ethanol solution was used as the colorimetric reaction substrate; then add 4.75 μL, 9.5 μL, 14.25 μL, 19 μL, 23.75 μL, 38 μL, 47.5 μL of 0.01 M peroxide Hydrogen solution, incubate at 40°C for 30 minutes to obtain hydrogen peroxide solutions with 50 μM, 100 μM, 150 μM, 200 μM, 250 μM, 400 μM, and 500 μM hydrogen peroxide solutions;...

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Abstract

The invention relates to a hydrogen peroxide colorimetric sensing method based on fluorescent polymer mimic enzyme, which belongs to the technical field of environment and biological analysis. The method comprises the following steps: concentrated sulfuric acid is added in diethylenetriamine drop by drop, and the materials are subjected to standing for 20-40 min under ice water-bath condition to obtain milk-white fluorescent polymer mimic enzyme; mimic enzyme dispersion liquid is added in a NaAc-HAc buffer solution, then 3,3',5,5'-tetramethyl benzidine ethanol solution is added in a reaction system as a colorimetric reaction substrate, an aqueous hydrogen peroxide solution is added to obtain a hydrogen peroxide solution with final concentration of hydrogen peroxide being 50 [mu]m-500 [mu]m, a light absorbance value of an ultraviolet-visible light area is determined, a concentration-light absorbance value standard curve is established, a solution with unknown hydrogen peroxide concentration is prepared, the light absorbance value of the solution is determined, and hydrogen peroxide concentration of the solution is calculated through the above standard curve, so that the hydrogen peroxide colorimetric sensing based on fluorescent polymer mimic enzyme is realized.

Description

technical field [0001] The invention belongs to the technical field of environmental and biological analysis, and in particular relates to a hydrogen peroxide colorimetric sensing method based on a fluorescent polymer imitating enzyme, and the imitating enzyme has good response characteristics to hydrogen peroxide. Background technique [0002] With the development of industrial and agricultural production and the improvement of urban living standards, environmental pollution and human health problems have become more and more prominent. As a typical oxidant, hydrogen peroxide is not only widely used in industrial and agricultural production processes, but also commonly used in industrial processes such as organic synthesis and chemical production. Especially hydrogen peroxide has a strong oxidizing property, and the large-scale use of hydrogen peroxide has brought serious harm to human health and the production and living environment. Therefore, the accurate and continuous...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31G01N21/64G01N21/78
CPCG01N21/3103G01N21/6402G01N21/78
Inventor 刘森张彤费腾
Owner JILIN UNIV
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