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Updated: Aug 18, 2026
Rapid Quantification of Oxidized and Reduced Forms of Glutathione Using Ortho -phthalaldehyde in Cultured Mammalian Cells In Vitro
Published on: June 28, 2024
Tunable Triazole-Dansyl Fluorescent Probes for Glutathione Detection: Modulating Probe-Biomarker Interactions for
Geethanjali Anand1, Karthika Kannan1, Jaseela Keedakkadan1
1Department of Chemistry, Indian Institute of Technology Palakkad, Palakkad, Kerala678623, India.
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A structurally tunable library of six triazole-dansyl fluorophores was investigated for substitution-dependent recognition of glutathione (GSH). All probes showed fluorescence quenching upon GSH addition, with the aliphatic-aliphatic derivative P4 exhibiting the highest sensitivity (LOD: 123 nM). High selectivity toward GSH was observed over competing amino acids, metal ions, and biomolecules, with minimal interference under competitive conditions. Mechanistic studies (UV-Vis, ESI-MS, 1H NMR, FTIR, PXRD, molecular docking, and DFT) revealed multivalent noncovalent interactions with a 2:1 binding stoichiometry, inducing perturbation of the intramolecular charge transfer (ICT) state. Time-resolved fluorescence confirmed a dynamic quenching pathway, while quantum yield measurements showed up to 49% quenching. pH- and temperature-dependent studies established optimal performance under physiological conditions. Practical applicability was demonstrated in human serum, cow urine, and goat blood with recoveries of 95-115% and negligible matrix effects. P4 did not exhibit any significant cytotoxicity in HeLa cells even at 100 μM. Fluorescence-based detection of GSH in HeLa cells and A549 cells confirmed the probe's sensing capability in mammalian cells, while cellular imaging established its suitability for bioimaging applications.
