Designing an ESIPT-based fluorescent probe for imaging of hydrogen peroxide during the ferroptosis process

Hydrogen peroxide (H2O2), depending on its levels, plays a crucial role in either modulating various biological processes as a signal molecule, or mediating oxidative damage as a toxin. Therefore, monitoring intracellular H2O2 levels is pivotal for exploring its physiological and pathological roles. Using a modified 2-(2′-hydroxyphenyl) benzothiazole (HBT) as the fluorophore, and a pinacol phenylborate ester as the responsive group, herein we developed an excited-state intramolecular proton transfer (ESIPT)-based probe BTFMB. The probe exhibited turn-on fluorescence response, large Stokes shift (162 nm) and low detection limit (109 nM) toward H2O2, and was successfully applied for monitoring exogenous and endogenous production of H2O2, and identifying accumulation of H2O2 during the ferroptosis process.