Nano copper oxide analogue enzyme and method for measuring hydrogen peroxide by using nano copper oxide analogue enzyme as peroxide analogue enzyme

A nano-copper oxide and peroxidase technology, applied in the field of analytical chemistry, can solve the problems of difficult purification, high price, limited natural enzyme sources, etc., and achieve the effects of simple preparation process, good dispersion stability and strong catalytic ability.

Inactive Publication Date: 2011-04-20
FUJIAN MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the source of natural enzymes is limited, difficult to purify, expensive, and in order to maintain its activity, th

Method used

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  • Nano copper oxide analogue enzyme and method for measuring hydrogen peroxide by using nano copper oxide analogue enzyme as peroxide analogue enzyme
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  • Nano copper oxide analogue enzyme and method for measuring hydrogen peroxide by using nano copper oxide analogue enzyme as peroxide analogue enzyme

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Experimental program
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Effect test

Embodiment 1

[0029] The preparation steps of nano copper oxide mimic enzyme are as follows:

[0030] (1) Get 150ml of copper nitrate solution of 0.02mol / L and 0.5ml glacial acetic acid and join in the three-necked bottle that condenser tube is housed, stir and be heated to boiling;

[0031] (2) Quickly add 10ml of 0.04g / ml sodium hydroxide solution, after adding, continue to stir for 5 minutes to obtain black copper oxide precipitate;

[0032] (3) centrifuging the black copper oxide precipitation obtained by the reaction, washing with absolute ethanol three times, and drying under reduced pressure to obtain a diameter of 6nm nano-copper oxide powder ( figure 1 It is the transmission electron micrograph of the nano-copper oxide prepared by the present invention), and the productive rate is 25%. The XRD characterization results of nano-copper oxide are as follows: figure 2 As shown, it can be seen from the figure that all the diffraction peaks of the product can be identified as monoclin...

Embodiment 2

[0035] The characteristic characterization steps of nano-copper oxide mimic enzyme are as follows:

[0036] (1) Take 3.9ml of double-distilled water and place it in a super constant temperature water bath at 37°C for 20 minutes;

[0037] (2) Add 850 μL of 30% hydrogen peroxide solution, 50 μL of 0.4 mg / ml nano-copper oxide mimic enzyme solution prepared in Example 1, 5.5 mg / ml 3,3',5,5'-tetramethylbis 200 μL of aniline hydrochloride solution, shake well, react at 37°C for 10 minutes, then take out;

[0038] (3) Using double distilled water as a blank, measure the absorption spectrum in the wavelength range of 400-800nm.

[0039] see test results Figure 5 , Figure 6 shown, from Figure 5 , Figure 6 It can be seen from the data that nano-copper oxide can catalyze the reaction of hydrogen peroxide to oxidize the peroxidase substrate 3,3',5,5'-tetramethylbenzidine hydrochloride, and has excellent characteristics of mimicking peroxidase .

Embodiment 3

[0041] Nano-copper oxide is used as a peroxide mimetic enzyme to measure hydrogen peroxide as follows:

[0042] (1) Preheat the reaction reagents (6 mmol / L phenol solution, 6 mmol / L 4-aminoantipyrine solution, pH 6.0 Tris-HCl buffer solution) at 37°C for 20 minutes;

[0043] (2) In the same centrifuge tube, add successively 2.0ml of the above-mentioned phenol solution, 790 μl of 4-aminoantipyrine solution, 960 μl of Tris-HCl buffer solution, and 300 μl of hydrogen peroxide solution of different concentrations, prepared in Example 1 0.2mg / ml nano-copper oxide simulated enzyme solution 50μl, after mixing, react at a constant temperature of 37°C for 20 minutes; 0.8, 0.9, 1.0mmol / L;

[0044] (3) Measure the absorbance at the maximum absorption wavelength of 505 nm with an ultraviolet-visible spectrophotometer.

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Abstract

The invention discloses a nano copper oxide analogue enzyme and a method for measuring hydrogen peroxide by using the nano copper oxide analogue enzyme as a peroxide analogue enzyme. The nano copper oxide analogue enzyme is nano copper oxide powder with a grain diameter ranging from 5 to 8 nanometers; the nano copper oxide is Cuo of a monoclinic system, the space group is C2/c; and the maximum absorption peaks of the typical ultraviolet-visible absorption spectra of the prepared nano copper oxide are at 280-nanometer positions. The nano copper oxide analogue enzyme is obtained by a chemical precipitation process. The nano copper oxide analogue enzyme has a catalytic property similar to that of peroxidase. When the nano copper oxide analogue enzyme is used in a 4-aminoantipyrine-ol system, the colorimetric detection of hydrogen peroxide can be realized, the hydrogen peroxide detection limit is 0.02mmol/L, and the linear range is 1mmol/L.

Description

technical field [0001] The invention relates to nano copper oxide and a method for measuring hydrogen peroxide as a peroxide mimetic enzyme, belonging to the technical field of analytical chemistry. Background technique [0002] Enzymes are efficient biocatalysts, and almost all reactions in organisms involve enzymes. Enzymatic analysis is widely used in clinical diagnosis, biology, food, chemistry, environment and other fields. Enzyme-catalyzed reactions have attracted much attention due to their high efficiency, specificity, and mild conditions. However, the source of natural enzymes is limited, difficult to purify, expensive, and in order to maintain its activity, the experimental conditions and operating environment are relatively harsh, which greatly restricts its application. s concern. [0003] The reported mimetic enzymes mainly include peroxide mimetic enzymes, cytochrome P450 mimetic enzymes, serine protease mimetic enzymes, phosphodiesterase mimetic enzymes, di...

Claims

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Application Information

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IPC IPC(8): C01G3/02B82Y40/00G01N21/31
Inventor 陈伟刘爱林李光文林新华
Owner FUJIAN MEDICAL UNIV
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