研究目的
Developing a coumarin-based dual optical probe for the selective detection of homocysteine (Hcy) with rapid response time, high sensitivity, and selectivity over other amino acids, especially cysteine (Cys) and glutathione (GSH).
研究成果
The coumarin-based probe DC demonstrates rapid, sensitive, and selective detection of Hcy through fluorescence and chromogenic changes. Its application in living cells and real samples highlights its potential for practical use in biological systems. The study provides a foundation for further research into selective thiol detection methods.
研究不足
The probe's response to Hcy is less effective in acidic conditions (pH<6), limiting its application in certain biological environments. Additionally, the probe shows significant responses to 2-mercapto ethanol and relatively weaker responses to 3-mercapto propionic acid, which may affect selectivity in complex samples.
1：Experimental Design and Method Selection:
The study utilized a coumarin-based probe (DC) for the selective detection of Hcy through fluorescence and chromogenic changes. The methodology involved the synthesis of the probe and its application in detecting Hcy in aqueous solutions and living cells.
2：Sample Selection and Data Sources:
The experiments were conducted using Hcy, Cys, GSH, and other amino acids in phosphate buffer solution (PBS) at physiological pH. Human serum and urine samples were also tested for practical application.
3：List of Experimental Equipment and Materials:
The probe DC was synthesized according to previous work. Fluorescence and UV-vis spectra were collected using a PerkinElmer LS-55 spectrometer and PerkinElmer Lambda 950 spectrophotometer, respectively. Confocal experiments were performed with a Leica TCS SPE confocal scanning microscope.
4：Experimental Procedures and Operational Workflow:
The probe's response to Hcy was evaluated through absorption and fluorescence titration experiments. The cytotoxicity of DC was tested by the standard MTS assay, and cell-imaging experiments were conducted with confocal fluorescence microscopy.
5：Data Analysis Methods:
The fluorescence intensity changes were analyzed to determine the probe's sensitivity and selectivity. The detection limit was calculated based on the standard deviation of the blank solution and the slope of the calibration plot.
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