研究目的
Designing and synthesizing two novel rhodamine-based fluorescent probes for the rapid and sensitive detection of Fe3+ ions in biological and environmental systems.
研究成果
RBA2 and RBA3 are effective 'turn-on' fluorescent probes for Fe3+ with high sensitivity, selectivity, and low detection limits. They form 1:1 complexes with Fe3+, as confirmed by multiple techniques, and are applicable in biological imaging and environmental monitoring, with RBA3 showing stronger binding due to sulfur atom coordination.
研究不足
The probes show slight interference from some metal ions like Hg2+, Al3+, Cr3+, and Pb2+ for RBA2, and Hg2+, Al3+, Cr3+, and Cu2+ for RBA3; the study is limited to in vitro and cell culture conditions, and further in vivo applications may require optimization.
1：Experimental Design and Method Selection:
The study involved designing and synthesizing rhodamine-based probes RBA2 and RBA3, using fluorescence and UV-vis spectroscopy for detection, and DFT calculations for theoretical analysis.
2：Sample Selection and Data Sources:
Probes were synthesized from rhodamine B; various metal ions were used for selectivity tests; Hela cells and real water samples (drinking water, tap water, Pearl River water) were used for application tests.
3：List of Experimental Equipment and Materials:
Equipment included AVANCE III HD 400 NMR spectrometer (Bruker), UV-2450 UV-Vis spectrophotometer (Shimadzu), F-4500 fluorescence spectrophotometer (Hitachi), Zeiss LSM 710 confocal microscope; materials included rhodamine B, various salts (e.g., FeCl3), solvents (e.g., DMSO), silica gel (200-300 mesh, Qingdao Haiyang Chemical Co., Ltd.).
4：Experimental Procedures and Operational Workflow:
Synthesis of probes via multi-step reactions; preparation of solutions; fluorescence and UV-vis titrations with Fe3+ and other ions; pH effect studies; reversibility tests with ethylenediamine; Job's plot and ESI-MS for stoichiometry; 1H NMR titrations; DFT calculations; cell culture and imaging; MTT assays for cytotoxicity; real water sample testing.
5：Data Analysis Methods:
Fluorescence intensity measurements; calculation of detection limits and binding constants using Benesi-Hildebrand plot; statistical analysis for cell viability; DFT calculations with Gaussian 09 using B3LYP functional and 6-31+G(d,p) basis set.
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NMR Spectrometer
AVANCE III HD 400
Bruker
Recording 1H NMR and 13C NMR spectra for structural confirmation of synthesized compounds.
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UV-Vis Spectrophotometer
UV-2450
Shimadzu
Recording UV-vis absorption spectra for detection of Fe3+ ions.
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Fluorescence Spectrophotometer
F-4500
Hitachi
Recording fluorescence responses for detection of Fe3+ ions.
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Confocal Microscope
LSM 710
Zeiss
Imaging Hela cells for fluorescence detection of Fe3+ in living cells.
ZEISS LSM 990 Spectral Multiplex
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Silica Gel
200-300 mesh
Qingdao Haiyang Chemical Co., Ltd.
Used in column chromatography for purification of synthesized compounds.
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