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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

Active Publication Date: 2018-11-13
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there are no domestic and foreign literatures and patent reports on the detection of choline by the ratiometric fluorescent probe method and the ratiometric fluorescent probe based on the copper nanocluster complex.

Method used

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  • Method for detecting choline by using ratio fluorescent probe based on copper nano-cluster compound
  • Method for detecting choline by using ratio fluorescent probe based on copper nano-cluster compound

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention belongs to the technical field of preparation of heavy metal nano materials and ratio fluorescent probes, and particularly relates to a stable copper nano-cluster compound based on lysozyme and a preparation method of a ratio fluorescent probe. Hydrazine hydrate serves as a reducing agent, lysozyme serves as a stabilizer, and copper nano cluster is prepared. Choline oxidase catalyzescholine to generate hydrogen peroxide, and hydrogen peroxide and horse radish peroxidase oxidize o-phenylenediamine to generate diaminophenolazine. Because an internal filter effect occurs between diaminophenolazine and copper nano cluster, the fluorescence of diaminophenolazine is gradually increased along with increasing of the concentration of choline, while the fluorescence of copper nano cluster is gradually quenched by diaminophenolazine, then a linear relation between the fluorescence peak ratio of copper nano cluster and diaminophenolazine and the choline concentration can be constructed, in this way, the ratio fluorescent probe is prepared. The preparation method of the probe is simple, the cost is low, the sensitivity of the product is high, the probe can be developed to be a novel ratio fluorescent probe and used for high-efficiency detection of choline in biological samples.

Description

Technical field: [0001] The invention belongs to the technical field of preparation of noble metal nanomaterials and ratio fluorescent probes, in particular to a preparation method based on lysozyme-stabilized copper nanocluster complexes and ratio fluorescent probes, and the prepared probes can be used for the detection of choline Efficient detection. Background technique: [0002] Choline is an essential nutrient for mammals, and choline derivatives play an important role in the development of human health. Choline's precursor, acetylcholine, is a neurotransmitter that is critical to the function of the nervous system and major organs throughout the body. Choline can also act as a precursor of the methyl donor glycine betaine and participate in one-carbon metabolism. Choline deficiency can lead to liver damage, neurological disorders, cell death, and cancer. Dairy products contain choline, and the nutritional intake of infants and young children is generally limited to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64C09K11/06C09K11/58
CPCC09K11/06C09K11/58C09K2211/1044G01N21/6428G01N2021/6432
Inventor 桂日军卜祥宁金辉付永鑫
Owner QINGDAO UNIV
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