Lighting up hydrogen peroxide in living cells by a novel quinoxalinamine based fluorescent probe

Hydrogen peroxide (H₂O₂), a member of small-molecule reactive oxygen species (ROS), plays vital roles in normal physiological activities and the occurrence of many diseases. In this work, two off-on fluorescent probes, QX8A-H₂O₂ and QX9A-H₂O₂, were firstly designed for H₂O₂ detection with novel fused quinoxalines as the fluorophores and boronate moiety as the reaction sites. By comparing the optical properties, QX9A-H₂O₂ with better performance was selected for further studies. QX9A-H₂O₂ exhibited a high sensitivity to H₂O₂ with the detection limit as low as 46 nM, and displayed a good selectivity towards H₂O₂ over other reactants such as ROS, biothiols and various ions. The detection was based on the intramolecular charge transfer (ICT), proceeding through a sequential oxidative hydrolysis, 1,6-rearrangement elimination and decarboxylation process to release the fluorophore QX9A. Moreover, probe QX9A-H₂O₂ was cell permeable and was successfully employed in both exogenous and endogenous H₂O₂ imaging in living cells.