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
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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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