Method for detecting hydrogen peroxide or glucose based on enzyme simulation by ferroferric oxide magnetic nanometer particle

A technology of ferroferric oxide and nanoparticles, which is applied in the field of analytical chemistry, can solve the problems of time-consuming and labor-intensive preparation and purification, and the variability of natural enzymes, and achieve good selectivity, sensitivity, and good sensitivity.

Inactive Publication Date: 2009-03-18
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, natural enzymes are volatile, easily degraded by proteases, and time-consuming and laborious to prepare and purify

Method used

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  • Method for detecting hydrogen peroxide or glucose based on enzyme simulation by ferroferric oxide magnetic nanometer particle
  • Method for detecting hydrogen peroxide or glucose based on enzyme simulation by ferroferric oxide magnetic nanometer particle
  • Method for detecting hydrogen peroxide or glucose based on enzyme simulation by ferroferric oxide magnetic nanometer particle

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The preparation steps of ferric oxide nanoparticle mimic enzyme are as follows:

[0038] (1) Mix 50mL concentration of 1M ferric chloride aqueous solution with 10mL concentration of 2M ferrous chloride hydrochloric acid solution (2M), and nitrogen gas for deoxygenation for 10 minutes;

[0039] (2) While vigorously stirring, the above-mentioned mixed solution was added dropwise to 500 mL of ammonia solution with a concentration of 0.7 M. The ammonia solution was deoxygenated and protected by nitrogen, and reacted at room temperature for 30 minutes, and the obtained black ferric oxide precipitated;

[0040] (3) Centrifuge the black iron ferric oxide precipitate obtained from the reaction, wash it with water three times; redisperse it in water to obtain ferric oxide nanoparticle-mimicking enzyme, and store it at room temperature for future use.

Embodiment 2

[0042] The steps for colorimetric detection of hydrogen peroxide by using iron ferric oxide nanoparticles to simulate enzymes are as follows:

[0043] (1) 24 μL of 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt with a concentration of 60 mM, 10 μL ferric oxide nanoparticle mimic enzyme stock solution and 24 μL of 100 mM HO 2 o 2 Add to 185 μL of acetic acid buffer solution with a concentration of 0.2M and pH=4.0;

[0044] (2) React the mixed solution obtained in (1) in a water bath at 45°C for 10 minutes;

[0045] (3) removing the iron ferric oxide nanoparticle simulative enzyme in the reaction solution with an external magnetic field to obtain a reaction solution for removing the ferric oxide nanoparticle simulative enzyme;

[0046] (4) Add 100 μL of the reaction solution obtained in (3) to 900 μL of water, mix well, and measure the absorption spectrum.

Embodiment 3

[0048] The steps for colorimetric detection of glucose by using iron ferric oxide nanoparticles to simulate enzymes and glucose oxidase are as follows:

[0049] (1) Mix 20 μL of 20 mg / mL glucose oxidase with 200 μL of glucose solutions of different concentrations, and react at 37° C. for 30 minutes; the concentration of the glucose solution is 5×10 -5 , 1×10 -4 , 5×10 -4 , 1×10 -3 , 5×10 -3 and 1×10 -2 mol / L;

[0050] (2) Mix 24 μL of 60 mM 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, 10 μL ferroferric oxide nanoparticle mimic enzyme stock solution and 800 μL concentration A 0.2M pH=4.0 acetate buffer solution was added to the 220 μL glucose reaction solution in the above step (1);

[0051] (3) The mixed solution in the above step (2) was reacted in a water bath at 45°C for 10 minutes, and the ferric oxide nanoparticles in the reaction solution were removed with an external magnetic field;

[0052] (4) Take 900 μL of the final reaction solutio...

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Abstract

The invention relates to a method for detecting hydrogen peroxide or glucose, based on ferroferric oxide nanometer particle mimetic enzyme, wherein the ferroferric oxide nanometer particle mimetic enzyme is obtained by co-precipitation method, the ferroferric oxide nanometer particle mimetic enzyme has the catalysis property similar to peroxidase, and the ferroferric oxide nanometer particle mimetic enzyme can be used to detect hydrogen peroxide for real-time presenting the colorimetric detection of hydrogen peroxide. The detected limit of hydrogen peroxide is 3x10<-6>mol/L, the linear range is 5x10<-6> to 1x10<-4>mol/L. The ferroferric oxide nanometer particle mimetic enzyme can be combined with glucose oxidase to realize the colorimetric detection of glucose. The detection limit of glucose is 3x10<-5>mol/L and the linear range is 5x10<-5> to 1x10<-3>mol/L. The colorimetric detection of glucose has high selectivity and sensitivity, without complex instruments.

Description

technical field [0001] The invention relates to a method for detecting hydrogen peroxide or glucose based on ferric iron tetroxide magnetic nanometer particle imitation enzyme, and belongs to the technical field of analytical chemistry. Background technique [0002] Natural enzymes have high substrate specificity and high catalytic efficiency under mild conditions. However, natural enzymes are volatile, easily degraded by proteases, and time-consuming and labor-intensive to prepare and purify. Therefore, people are trying to find various substitutes (ie, mimic enzymes) for natural enzymes. The simulated enzymes found so far are: artificial ribozyme and deoxyribozyme, cytochrome P450 simulated enzyme, serine protein kinase simulated enzyme, dioxygenate simulated enzyme, superphosphodiester oxygen simulated enzyme, linkase simulated enzyme, nuclease simulated Enzyme and peroxide mimetic enzymes, etc. Among them, the peroxide mimetic enzymes include: heme, heme, hemoglobin, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/78
Inventor 魏辉汪尔康
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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