A method for detecting reducing biomolecules by manganese dioxide nanosheets simulating oxidases

A technology of manganese dioxide and biomolecules, applied in the fields of nano-biomaterials and analytical chemistry, can solve problems such as variability, inactivation, and poor stability of natural enzymes

Active Publication Date: 2018-08-07
XI AN JIAOTONG UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

Therefore, the study of simulated enzymes can solve the problems of poor stability, variability and inactivation of natural enzymes.

Method used

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  • A method for detecting reducing biomolecules by manganese dioxide nanosheets simulating oxidases
  • A method for detecting reducing biomolecules by manganese dioxide nanosheets simulating oxidases
  • A method for detecting reducing biomolecules by manganese dioxide nanosheets simulating oxidases

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Embodiment 1

[0038] Detection of reducing biomolecules-glutathione GSH based on the method of the present invention

[0039](1) Add 100 μL of GSH solution of known concentration to 100 μL of manganese dioxide nanosheet solution and 0.5 mL of NaAc-HAc (0.2M) solution for 10 min at room temperature;

[0040] (2) Add the mixed solution in (1) to 200 μL TMB solution (1.0 mM), and react at room temperature for 5 min;

[0041] (3) Measure the absorbance value of the solution in (2) at 650 nm with a double-beam ultraviolet-visible spectrophotometer, and denote it as A. Use ultrapure water instead of GSH solution as a blank experiment, and its absorbance value at 650nm is recorded as A 0 .

[0042] (4) Absorbance net signal △A=A 0 -A is used as a quantitative indicator to draw the GSH standard curve to obtain a linear equation (refer to figure 2 ) and detection of GSH linear range (1μM-15μM) and detection limit (0.3μM).

[0043] (5) repeat above-mentioned steps (1), (2), (3) with GSH solutio...

Embodiment 2

[0046] Detection of reducing biomolecules-ascorbic acid AA based on the method of the present invention

[0047] (1) Add 100 μL of AA solution of known concentration to 100 μL of manganese dioxide nanosheet solution and 0.5 mL of NaAc-HAc (0.2M) solution for 10 min at room temperature;

[0048] (2) Add the mixed solution in (1) to 200 μL TMB solution (1.0 mM), and react at room temperature for 5 min;

[0049] (3) Measure the absorbance value of the solution in (2) at 650 nm with a double-beam ultraviolet-visible spectrophotometer, and denote it as A. Replace the AA solution with ultrapure water as a blank experiment, and record the absorbance value at 650nm as A 0 .

[0050] (4) Absorbance net signal △A=A 0 -A is used as a quantitative index, and the AA standard curve is drawn to obtain a linear equation (refer to image 3 ) and detection AA linear range (3-100μM) and detection limit (0.8μM).

[0051] (5) Repeat the above steps (1), (2) and (3) with the AA solution to be ...

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Abstract

A method for detecting reductive biomolecules by manganese dioxide nanosheets mimicking oxidases, manganese oxide nanosheets mimicking oxidases can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine TMB, 2,2 -Aziridine-bis(3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt ABTS and o-phenylenediamine OPD and other substrates, making them change from colorless to blue, green and orange respectively, and at the same time , manganese dioxide nanosheets can sensitively and selectively undergo redox reactions with reducing biomolecules such as glutathione and ascorbic acid, thereby changing the concentration of oxidation products of substrates such as TMB, ABTS and OPD. The analytical method quantitatively detects reducing biomolecules such as glutathione and ascorbic acid; this method has the characteristics of simple operation, high sensitivity, good reproducibility and high selectivity; its linear range for the detection of glutathione is 1‑15 μM, The detection limit is 0.3 μM; the detection linear range of ascorbic acid is 3‑100 μM, and the detection limit is 0.8 μM. This method can also be used for the detection of various polyphenolic compounds.

Description

technical field [0001] The invention belongs to the technical field of nano-biological materials and analytical chemistry, and in particular relates to a method for detecting reducing biomolecules by simulating oxidases with manganese dioxide nanosheets. Background technique [0002] Enzymes are macromolecular substances with biocatalytic functions, which have strong substrate specificity and high catalytic efficiency under mild conditions. However, the extraction and purification process of natural enzymes is time-consuming and cumbersome, the cost is high, and the stability is poor. At the same time, the catalytic activity of natural enzymes is easily denatured and inactivated by the environment, which severely limits their further applications. Mimetic enzymes are substances with similar functions to natural enzymes, and can still maintain high stability under harsh conditions. Therefore, the study of mimic enzymes can solve the problems of poor stability, variability an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N31/10G01N21/78G01N21/33
CPCG01N21/33G01N21/78G01N31/10
Inventor 孟令杰刘晶党东锋曹瑞军
Owner XI AN JIAOTONG UNIV
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