Preparation of protamine gold nanoclusters and application in analogue enzyme color comparison and fluorescence detection

A technology of protein gold and nanoclusters, which is applied in the field of materials science and analytical chemistry, can solve the problems such as the type, quantity and sensitivity of nanocluster materials that cannot be satisfied, and achieve short fluorescence lifetime, superior fluorescence and chemical stability, The effect of strong luminous performance

Inactive Publication Date: 2017-04-26
NANHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although important progress has been made in the research of metal nanocluster materials, the reported nanocluster materials still cannot meet the needs of chemical (biological) sensing, environment and biomedical fields in terms of type, quantity, sensitivity and specificity, especially those that can be used There are few reports on nanocluster materials that can be used as fluorescent dyes and have enzyme-mimicking activity

Method used

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  • Preparation of protamine gold nanoclusters and application in analogue enzyme color comparison and fluorescence detection
  • Preparation of protamine gold nanoclusters and application in analogue enzyme color comparison and fluorescence detection
  • Preparation of protamine gold nanoclusters and application in analogue enzyme color comparison and fluorescence detection

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Synthesis of multifunctional protamine gold nanoclusters specifically comprises the following steps:

[0038] (1) The glassware used in the experiment were all pre-washed with HCl:HNO 3 The prepared aqua regia was soaked overnight, and then thoroughly washed with ultrapure water;

[0039] (2) Prepare 0.02g / mL HAuCl 4 stock solution and 0.625-2.5 mg / mL protamine sulfate solution (PS); HAuCl 4 Dilute the solution to 10mmol / L when used;

[0040] (3) Pipette 10-50mL 10mmol / L HAuCl 4 Put the solution in 10-50mL 0.625-2.5mg / mL PS solution, stir vigorously in a water bath at 37°C for 2 minutes, add 1mol / L NaOH solution to adjust the pH to 12, and keep stirring the mixed solution at 25-50°C for 8 ~24 hours;

[0041] (4) The prepared PS-AuCNs were stored at 4°C in the dark.

[0042] Characterization and performance investigation of multifunctional protamine gold nanoclusters obtained in the present invention. The sample is selected from the multifunctional p...

Embodiment 2

[0047] Embodiment 2: Determination of mercury ions based on protamine gold nanoclusters (PS-AuCNs) catalytic colorimetric method,

[0048]

[0049] Mercury ion determination result and recovery rate determination in table 1 water sample

[0050] (8) Sample analysis, respectively get laboratory tap water (sample 1), water from Nanhua University Chitang (sample 2), Xiangjiang water (sample 3) a ,3 b ,3 c ). First filter twice with quantitative filter paper and place in a beaker, heat and boil on an electric stove, keep boiling for 10 minutes, take it off, let it cool naturally, precipitate, and filter with a 0.22μm filter membrane to be tested. The test results of the samples are shown in Table 1.

[0051] (7) Under the best experimental conditions, according to the substances that may coexist in the environmental water samples, the influence of various interfering substances on the measurement results was tested. When the relative error is controlled at ± 5%, 1000 times ...

Embodiment 3

[0058] Example 3: Detection of uranyl ions (UO) based on protamine gold nanoclusters (PS-AuCNs) fluorescence method 2 2+ ),

[0059] (1) Pipette 30-70 μL of protamine gold nanoclusters (PS-AuCNs) into an EP tube, add appropriate amount of NaAc-HAc buffer (pH5.5) and different concentrations of UO 2 2+ Standard solution, dilute to 500 μL with sterilized water, mix thoroughly, and react for 35 minutes;

[0060] (2) Pipette an appropriate amount of solution obtained in step (1). On the F-4500 fluorescence spectrophotometer, set the excitation and emission slits to be 5cm and 10cm respectively, and the negative voltage of the photomultiplier tube to be 700V. Scanning in the wavelength range (λ ex =300nm), at λ em =599nm measures the fluorescence change value of the system, and draws a standard curve;

[0061] (3) The present invention can be used for measuring uranyl ion (UO 2 2+ ) concentration range is 2.0×10 -8 mol / L~1.0×10 -5 mol / L; detection limit is 6.1×10 -9 mol ...

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Abstract

The invention relates to preparation of protamine gold nanoclusters and application in analogue enzyme color comparison and fluorescence detection. The protamine gold nanoclusters not only have activity of horseradish peroxidase and oxidase, but also have fluorescent characteristic. Hereby, a protamine gold nanocluster analogue enzyme visual color comparator and a fluorescence sensor are established respectively and are used for detecting heavy metal ions. The protamine gold nanoclusters are prepared simply and conveniently, are low in cost, and have excellent fluorescence stability and chemical stability, and the shortcomings that fluorescence of traditional dyestuff is short in service life, photobleaching occurs easily and the like can be overcome; a fluorescence method is high in sensitivity and good in stability, and expensive instruments are not required; and analogue enzyme catalytic color comparison is sensitive and environment-friendly, and can be used for field real-time monitoring. The protamine gold nanoclusters are expected to be expanded and applied to detection and analysis of related substances in the fields of environment and biomedicines and disease markers, and have wide application prospect.

Description

technical field [0001] The invention relates to the fields of material science and analytical chemistry, in particular to the preparation of a protamine gold nanocluster and its application in simulated enzyme colorimetry and fluorescence detection. Background technique [0002] Metal nanocluster materials have attracted extensive research interest in recent years. This is because: on the one hand, fluorescent metal nanoclusters have incomparable advantages over traditional organic dyes, such as low toxicity, good photostability, large Stokes shift, high quantum yield and good biocompatibility; on the other hand, Metal nanocluster mimic enzymes are also favored by people. Compared with natural enzymes, artificial enzyme mimics have the advantages of easy modification, preparation, low price, strong specificity, and high sensitivity, which can overcome the limited source of natural enzymes and their susceptibility to environmental inactivation. and expensive disadvantages. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B22F9/24C09K11/58G01N21/64B82Y20/00B82Y40/00
CPCG01N21/643C09K11/58B82Y20/00B82Y40/00B22F9/24B22F1/0553B22F1/054
Inventor 王永生黄艳琴
Owner NANHUA UNIV
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