Method for detecting choline by using ratio fluorescent probe based on copper nano-cluster compound
A ratiometric fluorescent probe, copper nanocluster technology, applied in fluorescence/phosphorescence, chemical instruments and methods, measurement devices, etc., to achieve the effects of low cost, high product sensitivity, and simple preparation method
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Embodiment 1
[0020] The preparation process and principle of a method for detecting choline with a ratiometric fluorescent probe based on copper nanocluster complexes involved in this example are as follows: figure 1 As shown, the specific process steps are:
[0021] Preparation of copper nanoclusters: 2 mg of copper sulfate pentahydrate was added to 5 ml of twice distilled water to prepare an aqueous solution of copper sulfate. Under magnetic stirring, 0.3 g of lysozyme was dissolved in 10 ml of double distilled water until it was completely dissolved, and then the aqueous solution of lysozyme was placed in a constant temperature water bath at 45 degrees Celsius for incubation. Under magnetic stirring, the copper sulfate aqueous solution was slowly added to the lysozyme aqueous solution to prepare a mixed aqueous solution of lysozyme and copper sulfate, and stirred in a water bath at 45 degrees Celsius for 10 minutes. Add 1 mol / liter of sodium hydroxide solution to the mixed aqueous solu...
Embodiment 2
[0027] The specific process steps for the preparation of copper nanoclusters in this example are the same as those in Example 1. Weigh 162 mg of o-phenylenediamine and dissolve it in phosphate saline buffer to obtain a 3 mmol / L o-phenylenediamine solution. Measure 200 microliters of the o-phenylenediamine solution, add 10 microliters of 1.5 g / L horseradish peroxidase and 10 microliters of 5-20 mmol / L aqueous hydrogen peroxide solution. The prepared mixed solution was reacted in the dark at 37 degrees Celsius for 30 minutes, and then 40 microliters of 20 g / L copper nanoclusters dispersed in the prepared phosphate saline buffer solution were added, stirred slowly, and left at room temperature for 10 minutes to obtain copper Homogeneous solution of nanoclusters and diaminophenazines. Measure the fluorescence emission spectra of copper nanoclusters and diaminophenazine homogeneous aqueous solution at different molar concentrations of hydrogen peroxide, and fit the linear relation...
Embodiment 3
[0030] The specific process steps for the preparation of copper nanoclusters in this example are the same as those in Example 1. Weigh 162 mg of o-phenylenediamine and dissolve it in phosphate saline buffer to obtain a 3 mmol / L o-phenylenediamine solution. Measure 200 microliters of the o-phenylenediamine solution, add 10 microliters of 1.5 g / L horseradish peroxidase and 10 microliters of 10-50 mmol / L aqueous hydrogen peroxide solution therein. The prepared mixed solution was reacted in the dark at 37 degrees Celsius for 30 minutes, then added 25 g / L copper nanoclusters dispersed in 40 microliters of the prepared phosphate saline buffer solution, stirred slowly, and placed at room temperature for 10 minutes to obtain copper Homogeneous solution of nanoclusters and diaminophenazines. Measure the fluorescence emission spectra of copper nanoclusters and diaminophenazine homogeneous aqueous solution at different molar concentrations of hydrogen peroxide, and fit the linear relati...
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