Enhancing probe's sensitivity for peroxynitrite through alkoxy modification of dicyanovinylchromene

An intramolecular charge transfer (ICT)-based fluorescent probe P-ONOO^- was synthesized to detect ONOO^-. After responding to peroxynitrite, the dicyano-vinyl group of P-ONOO^- generates the aldehyde group, emitting strong green fluorescence accompanied by quenching of the yellow fluorescence. According to the calculated Fukui function, the modification of the alkoxy group can enhance the f⁺ of P-ONOO^-, which can enhance the probe's nucleophilic addition reactivity with ONOO^-. It has been experimentally verified that P-ONOO^- shows fast response (within 30 s), excellent sensitivity (the detection limit = 10.4 nM), and good selectivity towards ONOO^-. Additionally, the probe P-ONOO^- has high membrane permeability and good biocompatibility, which can image endogenous ONOO^- and exogenous ONOO^- in HeLa cells.