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An enzyme-free one-pot colorimetric detection method for glucose based on enzyme-like nanomaterials

A glucose and glucose oxidase technology, which is applied in the field of enzyme-free one-pot glucose color detection based on enzyme-like nanomaterials, can solve the problems of increased cost, difficult control, and complicated detection process, and achieve high temperature stability.

Active Publication Date: 2020-08-07
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be known from previous reports that the reported enzyme-like nanomaterials only have imitation peroxidase activity but no glucose oxidase activity, so the glucose chromogenic detection method constructed needs to be carried out in the presence of glucose oxidase. Complex and difficult to control, increased cost due to use of biological enzymes

Method used

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  • An enzyme-free one-pot colorimetric detection method for glucose based on enzyme-like nanomaterials
  • An enzyme-free one-pot colorimetric detection method for glucose based on enzyme-like nanomaterials
  • An enzyme-free one-pot colorimetric detection method for glucose based on enzyme-like nanomaterials

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] MnO based on BSA template 2 Preparation of NSs:

[0035] (1) Mix BSA (final concentration 100 µg / mL) and manganese acetate aqueous solution (final concentration 2 mM) and incubate for 1 h;

[0036] (2) Add an aqueous solution of sodium hydroxide to the above mixed solution and mix well, shake and react for 24 h;

[0037] (3) Centrifuge the above reaction solution, remove the supernatant containing unreacted components, collect the precipitate, and store it at 4°C for future use; BSA can also be collected and purified by dialysis or filtration.

Embodiment 2

[0039] MnO 2 Analysis of enzymatic activity of NSs:

[0040] MnO 2 Mimic peroxidase activity analysis of NSs: The catalytic reaction system is composed of H 2 o 2 (1 mmol / L), the MnO obtained in the above examples 2 NSs (25 µg / ml), organic chromogen TMB (0.5 mmol / L) in acetate buffer (pH 3.5). After reacting at room temperature (25 °C) for 30 minutes, the absorbance at 650 nm was detected with a microplate reader. Such as figure 2 As shown, with TMB as the color reagent, MnO 2 NSs exhibit mimic peroxidase activity (black curve). In another control experiment, the MnO in the above experiment 2 NSs were replaced with the same volume of water as figure 2 As shown, without MnO 2 NSs have no mimetic activity (grey curve).

[0041] In another two groups of experiments, the organic chromogen was replaced by OPD and ABTS, and after reacting at room temperature (25 °C) for 30 minutes, the absorbance value at 650 nm was detected by a microplate reader. Such as imag...

Embodiment 3

[0044] MnO 2 pH optimization of NSs mimic enzyme activity:

[0045] MnO 2 pH optimization of NSs imitation peroxidase activity: the catalytic reaction system is composed of H 2 o 2 (1 mmol / L), MnO 2 NSs (25 µg / ml), organic chromogenic agent TMB (0.5 mmol / L) with different pH buffers (pH 1.0−2.0, glycine-hydrochloric acid buffer; pH 3.0−6.0, acetic acid-sodium acetate buffer; pH 7.0−8.0, phosphate buffer; pH 9.0, Tris-HCl buffer). After reacting at room temperature (25 °C) for 10 minutes, the absorbance at 650 nm was detected with a microplate reader. Such as Figure 6 Shown, MnO 2 NSs exhibited peroxidase activity at slightly acidic pH (pH 3.0−5.0), and the optimum pH was around 3.5 (black curve).

[0046] MnO 2 pH optimization of NSs imitation glucose oxidase activity: the catalytic reaction system contains glucose (1 mmol / L), MnO 2 NSs (25 µg / ml) and buffers of different pH (pH 1.0−2.0, glycine-HCl buffer; pH 3.0−6.0, acetic acid-sodium acetate buffer; pH 7.0−8.0, ...

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Abstract

The invention relates to the field of nanomaterials, catalysis and analytical chemistry, in particular to a non-enzymatic one-pot glucose chromogenic detection method based on biomimetic nanomaterials. The method comprises the steps that bovine serum albumin is mixed with manganous ions by means of nanostructure characteristics of the bovine serum albumin and properties of functional groups of the bovine serum albumin, after sodium hydroxide is added, a normal temperature reaction is conducted, and then manganese dioxide nanosheets are generated under the stability of the bovine serum albumin. The material has the dual activities of imitation peroxidase and imitation glucose oxidase, and the optimal activity conditions of the two biomimetic activities are similar. Therefore, a novel one-step non-enzymatic glucose chromogenic detection method is established, and the material has a wide application prospect in the fields of analytical chemistry, environmental engineering and catalysis.

Description

technical field [0001] The invention relates to the fields of nanomaterials, catalysis and analytical chemistry, and specifically includes an enzyme-free one-pot glucose color detection method based on an enzyme-like nanomaterial. Background technique [0002] Enzyme-like nanomaterials are a class of nanomaterials with enzyme-like activity that can catalyze chemical reactions (non-electrochemical catalytic reactions). In recent years, compared with enzymes, enzyme-mimicking nanomaterials have been widely used in the field of catalysis due to their larger specific surface area, higher stability and activity. Manganese dioxide (MnO 2 ) As an important inorganic nanomaterial, it has received extensive attention. A series of studies have shown that MnO 2 Has good peroxidase activity. Lv et al. in "BSA-templatedMnO 2 Nanoparticles as both peroxidase and oxidase mimics" uses bovine serum albumin (BSA) as a template to synthesize MnO 2 nanoparticles, and explored their peroxid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/78G01N21/31
CPCG01N21/31G01N21/78G01N2201/129
Inventor 韩磊张海姣
Owner QINGDAO AGRI UNIV
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