研究目的
To design and synthesize a novel colorimetric fluorescent probe for selective detection of sulfur dioxide in SH-SY5Y neuroblastoma cells and its application in Traditional Chinese Medicines.
研究成果
The probe DTCC is highly effective for fast, sensitive, and selective detection of SO2 derivatives, with low cytotoxicity and good biocompatibility. It successfully images SO2 in living cells and quantifies it in TCM samples, demonstrating potential for biological and environmental monitoring.
研究不足
The intermediate aldehyde-hydrogen sulphite adduct is unstable in aqueous solution, limiting isolation and long-term stability. The probe's performance may be affected by complex matrices in real samples, and further optimization is needed for broader applications.
1:Experimental Design and Method Selection:
The study involved designing and synthesizing a coumarin-thiophene-based fluorescent probe (DTCC) for detecting SO2 derivatives through nucleophilic addition reactions. Theoretical calculations using DFT and TD-DFT were employed to understand optical properties.
2:Sample Selection and Data Sources:
SH-SY5Y neuroblastoma cells were used for cytotoxicity and imaging studies. Real TCM samples (e.g., Dioscorea opposite Thunb, Angelica sinensis, Lycium barbarum) were analyzed for SO2 content.
3:List of Experimental Equipment and Materials:
Instruments included UV-2550 spectrophotometer, F-7000 fluorescence spectrophotometer, FV1000 confocal microscope, Varian Gemini NMR spectrometer, micro TOF-QII mass spectrometer. Materials included various chemical reagents, solvents, and cell culture components.
4:Experimental Procedures and Operational Workflow:
Synthesis of DTCC in four steps, titration experiments with SO2 derivatives, cytotoxicity assays using CCK-8 kit, cell imaging with confocal microscopy, and application in TCM samples for SO2 detection.
5:Data Analysis Methods:
Fluorescence and UV-vis spectra analysis, linear regression for detection limits, statistical analysis for selectivity and pH dependence, and DFT calculations for mechanistic insights.
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UV-2550 spectrophotometer
UV-2550
Shimadzu
Determining UV-vis absorption spectra
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F-7000 fluorescence spectrophotometer
F-7000
Hitachi
Recording fluorescence spectra
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FV1000 confocal microscope
FV1000
Olympus
Cell fluorescence imaging
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micro TOF-QII mass spectrometer
micro TOF-QII
Bruker
Recording HRMS spectra
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Varian Gemini NMR spectrometer
Gemini 2000 DMX600
Varian
Obtaining NMR spectra
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Millipore water purification system
Millipore
Preparing deionized water
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Cell counting kit-8
CCK-8
Detecting cytotoxicity
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Microplate reader
Measuring absorbance in cytotoxicity assays
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