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A kind of nano enzyme and its preparation method for the detection of food antioxidant activity

A technology of antioxidant activity and nanozyme, applied in chemical instruments and methods, measurement of color/spectral characteristics, material analysis by observing the influence of chemical indicators, etc., can solve the limitation of large-scale application of ABTS method and cumbersome purification methods , Enzyme denaturation and other problems, to achieve excellent peroxidase-like activity, good practical application prospects, and good dispersibility

Active Publication Date: 2022-05-17
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the production of natural peroxidases involves several disadvantages, such as high cost, time-consuming, cumbersome purification methods, etc.
In addition, the catalytic activity of natural enzymes is easily affected by environmental conditions, such as extreme pH and temperature, which can cause significant enzyme denaturation, severely limiting the large-scale application of the ABTS method.

Method used

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  • A kind of nano enzyme and its preparation method for the detection of food antioxidant activity
  • A kind of nano enzyme and its preparation method for the detection of food antioxidant activity
  • A kind of nano enzyme and its preparation method for the detection of food antioxidant activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1A

[0034] The synthesis of embodiment 1Au@Cu-HCF nanozyme

[0035] Dissolve 18.4 mg of potassium hexacyanoferrate and 0.15 g of citric acid in 10 mL of deionized water, and mix well. Then, inject 0.2 μL of sodium tetrachloroaurate (III) (1 mg) under vigorous stirring, continue stirring for 2 min, and add anhydrous copper sulfate solution (4 mL, 5 mg / mL) dropwise to the above mixture until it becomes Wine red, Au@Cu-HCF nanozyme solution is obtained, the nanozyme solution is centrifuged at 17000 rpm for 15 minutes, separated to obtain Au@Cu-HCF nanozyme.

[0036]Gold-doped nanozyme particles were prepared by a one-step self-assembly method. Transmission electron microscope (TEM) analysis showed that the morphology of Au@Cu-HCF nanozyme particles was spherical with a diameter of about 15 nm ( figure 1 ). Energy dispersive X-ray spectroscopy results showed that the Au@Cu-HCF nanozyme was composed of Cu, Au, Fe, N and C ( figure 2 ).

Embodiment 2

[0037] Example 2 Preparation of PEG-SH modified Au@Cu-HCF nanozyme

[0038] After the wine red Au@Cu-HCF nanozyme solution prepared in Example 1 was continuously stirred for 12 hours under the condition of avoiding light, 50 mg COOH-PEG-SH was added, and the stirring was continued for 12 hours. After centrifugation for 15 minutes, the PEG-SH modified Au@Cu-HCF nanozyme was obtained.

[0039] In this embodiment, by utilizing the strong interaction between gold and sulfur, COOH-PEG-SH is used to coat nanoparticles, which are modified with PEG-SH, so as to further improve the stability and biocompatibility of nanoparticles.

Embodiment 3A

[0040] Example 3 Determination of Au@Cu-HCF Nanozyme Peroxidase Activity

[0041] Take 40 μL of the Au@Cu-HCF nanozyme solution prepared in Example 1, mix it with 40 μL of deionized water and add 100 μL of TMB buffer (TMB buffer A+B, buffer A: take 8.2 g of anhydrous sodium acetate, 2.5g of β-cyclodextrin and 0.4g of urea hydrogen peroxide were dissolved in 1L of water; buffer B: 100mg of TMB was dissolved in 10mL of DMSO) for reaction, and 100μL of H 2 SO 4 Terminate the reaction, and record the UV-Vis absorption spectrum of the TMB system in the range of 400nm-800nm ​​with a UV-Vis spectrophotometer.

[0042] Mix 50 μL of the Au@Cu-HCF nanozyme solution prepared in Example 1 with 50 μL of deionized water, and then add 50 μL of ABTS buffer (containing 4 mL of PBS, 200 μL of 3% H 2 o 2 and 10 mg ABTS) for color development, and after reacting for 10 min, record the UV-Vis absorption spectrum of the ABTS system in the range of 400nm-800nm ​​with a UV-Vis spectrophotometer. ...

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Abstract

The invention discloses a nanozyme used for detecting the antioxidant activity of food and a preparation method thereof, belonging to the field of nanobiotechnology. The reaction raw materials of the nanozyme of the present invention include potassium hexacyanoferrate, citric acid, sodium tetrachloroaurate (III) and copper sulfate, and the preparation method is: potassium hexacyanoferrate and citric acid are dissolved in water, and then sequentially Sodium tetrachloroaurate (III) and copper sulfate are added to react to obtain nanozyme. The nanozyme of the present invention is a gold-doped nanozyme Au@Cu-HCF, which can be used to detect hydrogen atom transfer (HAT) antioxidant substances and single electron transfer (SET) antioxidant substances. To measure the total antioxidant capacity in food, it has a good practical application prospect in the detection of TAC in plant-derived food.

Description

technical field [0001] The invention relates to the field of nanobiology technology, in particular to a nanozyme used for the detection of food antioxidant activity and a preparation method thereof. Background technique [0002] Natural antioxidant substances have the ability to reduce or scavenge free radicals. They can not only protect natural food from oxidative damage, but also neutralize oxidative stress in the human body and maintain the body's redox dynamic balance. The analysis and identification of natural antioxidant capacity of foods has received great attention in the field of nutrition and health. [0003] At present, traditional instrumental analysis methods including gas chromatography (GC) and liquid chromatography (LC) are widely used as an effective means to identify antioxidant substances, but such methods are time-consuming, have high professional requirements for operators, and require high equipment consumables. Expensive and not capable of portable an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J31/28B01J31/04B01J27/26B01J31/06B01J35/02B01J35/08G01N21/31G01N21/76G01N21/78
CPCB01J31/28B01J27/26B01J31/04B01J31/069B01J35/08B01J35/023B01J31/003G01N21/3151G01N21/76G01N21/78G01N2021/3155Y02A40/90
Inventor 张富源王向红桑亚新李雅梅李晓萌
Owner HEBEI AGRICULTURAL UNIV.
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