Method for rapidly fluorescently detecting hydrogen peroxide on basis of etched nano silver triangle
A hydrogen peroxide and fluorescence detection technology, which is applied in the direction of fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of low sensitivity, insufficient stability, and limited practical application, and achieve low cost, simple and fast detection, and high sensitivity. Effect
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[0033] (1) Preparation of nano-silver triangles. 40mL aqueous solution, including 400μL, 0.01M silver nitrate solution, 600μL, 0.1M trisodium citrate solution, 96μL, 30wt% H 2 o 2 Stir vigorously at room temperature for 10 min. Then, quickly inject 400 μL of 0.1 M sodium borohydride solution.
[0034] (2) Take 40mL nano-silver triangle solution. Add 200 μL bovine serum albumin solution (10 mg / mL) into the nano-silver triangular colloid solution, mix well, and let stand overnight. Add 300 μL of fluorescent dye (1 mg / mL) to the above mixed solution, mix well, and let stand for 4 h. The mixed solution was centrifuged at 8000rpm for 20min, resuspended with 0.015M trisodium citrate solution, and concentrated 10 times at the same time.
[0035] (3) Take 100uL of hydrogen peroxide of different concentrations (such as: 0, 0.025, 5, 30, 50, 100μM), add it to the suspension described in (1), shake and mix, let stand for 20min, and check the mixture The fluorescence intensity of th...
Embodiment 1
[0043] Embodiment 1 The detection method principle of this hydrogen peroxide is shown in the following formula:
[0044] 2Ag+H 2 o 2 →2Ag + +2OH -
[0045] The method is as follows:
[0046] (1) Preparation of nano-silver triangle-bovine serum albumin (BSA)-fluorescein isothiocyanate (FITC) complex. Take 40mL nano-silver triangle solution, add 200 μL bovine serum albumin solution (10mg / mL) into the nano-silver triangle colloid solution, mix well, and let stand overnight. Then 300 μL of fluorescein isothiocyanate (1 mg / mL) was added to the above mixed solution, mixed evenly, and allowed to stand for 4 h. The mixed solution was centrifuged at 8000rpm for 20min, resuspended with 0.015M trisodium citrate solution, and concentrated 10 times at the same time.
[0047] (2) Preparation of hydrogen peroxide standard curve. Draw 100uL of hydrogen peroxide solution of different concentrations (such as: 0, 0.025, 5, 30, 50, 100μM), add it to the suspension described in step (1), o...
Embodiment 2
[0050] Embodiment 2 The method is specifically as follows:
[0051] (1) Preparation of nano-silver triangle-bovine serum albumin-tetramethylrhodamine-5-isothiocyanate complex. Get 40mL nano-silver triangular colloidal solution. Add 200 μL bovine serum albumin solution (10 mg / mL) into the nano-silver triangular colloid solution, mix well, and let stand overnight. Then 300 μL of tetramethylrhodamine-5-isothiocyanate (1 mg / mL) was added to the above mixed solution, mixed evenly, and allowed to stand for 4 h. The mixed solution was centrifuged at 8000rpm for 20min, resuspended with 0.015M trisodium citrate solution, and concentrated 10 times at the same time.
[0052] (2) Take 100 uL of hydrogen peroxide solution of different concentrations (such as: 0, 0.025, 5, 30, 50, 100 μM), add it to the suspension described in step (1), oscillate and mix well, and let it stand for 20 minutes. The fluorescence intensity of the mixed reaction system was detected (excitation wavelength 546n...
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