Novel förster resonance energy transfer (FRET)-based ratiometric fluorescent probe for detection of cyanides by nucleophilic substitution of aromatic hydrogen (SNArH)

Abstract

The development of novel fluorescent probes for real-time detection of cyanides (CN⁻) in environmental and biological systems has become a significant focus in chemical sensing. Particularly, ratiometric fluorescence sensing offers a unique method for precise and quantitative detection of cyanides, even under complex conditions. We report herein the design of a new ratiometric fluorescent probe for cyanides based on modulation of Förster resonance energy transfer (FRET) coupled with novel cyanide-induced nucleophilic substitution of aromatic hydrogen (SNArH). The target probe (R1) is developed by introducing coumarin fluorophores as FRET donors into a 3-nitro-naphthalimide acceptor, which is easily synthesized and exhibits a colorimetric change from colorless to faint yellow and a significant ratiometric fluorescence shift (Δλ = 114 nm) upon cyanide binding. A clear ratiometric signal at I₅₈₂/I₄₆₈ was obtained, with a limit of detection of 5.69 μM. The sensing mechanism was confirmed through ¹H NMR titration and LC-MS analysis. Additionally, R1-loaded strips were easily prepared, serving as a portable device for detecting CN⁻ with visible color changes. The probe R1 has been successfully utilized for real-time monitoring of cyanide in food materials and water samples. Importantly, fluorescence bioimaging studies in HeLa cells were conducted, demonstrating the probe’s capability for ratiometric detection of exogenous CN⁻ in living systems.