Application of RET Approach for Ratiometric Response Enhancement of ICT Fluorescent Hg2+ Probe based on Crown‐containing Styrylpyridinium Dye

Styrylpyridinium dye bearing azadithia‐15‐crown‐5 ether receptor group SP and 4‐alkoxy‐1,8‐naphthalimide fluorophore were linked using copper‐catalyzed azide‐alkyne cycloaddition click reaction to afford dyad compound NI‐SP. Chemosensor NI‐SP exhibited selective ratiometric fluorescent response to the presence of Hg2+ in aqueous solution due to the interplay between resonance energy transfer (RET) and intramolecular charge transfer (ICT) processes occurred upon excitation. The observed switching of the ratio of emission intensities in the blue and red channels R was higher than in the case of monochromophoric styrylpyridine derivative SP showing ratiometric response based on ICT mechanism only. Biological studies revealed that NI‐SP penetrates into human lung adenocarcinoma A549 cells and accumulates in cytoplasm and lysosomes. When cells were pre‐incubated with mercury (II) perchlorate, the ratio R was increased 2.6 times, which enables detection of intracellular Hg2+ ions and their quantitative analysis in the 0.7–6.0 μM concentration range. Novel ratiometric chemosensor for Hg2+ cations based on «naphthalimide–styrylpyridine» RET pair has been designed and prepared. Ratiometric response of the probe was higher than that of the monochromophoric styrylpyridine chemosensor. Synthesized compound was capable of detection and quantification of Hg2+ content in living cells.