Development of ratiometric fluorescent probes based on peptides for sensing Pb²⁺ in aquatic environments and human...

The detection of Pb²⁺ ions in aquatic environments and biofluid samples is crucial for assessment of human health. Herein, we synthesized two fluorescent probes (1 and 2) consisting of the peptide receptor for Pb²⁺ and a benzothiazolyl-cyanovinylene fluorophore that exhibited excimer-like emission when it aggregated. The peptide-based probes sensitively detected Pb²⁺ in purely aqueous solution (1% DMF) through ratiometric fluorescent response with a decrease in monomer emission at 520 nm and an increase in excimer emission at 570 nm. Specially, probe 2 showed remarkable detection features such as high selectivity for Pb²⁺over 15 metal ions, high binding affinity (K_d = 5.83 × 10^-7 M) for Pb²⁺, significant emission intensity changes, low detection limit (3.8 nM) of Pb²⁺, high water solubility, and visible light excitation (450 nm). Probe 2 was successfully used to quantify nanomolar concentration (0 ∼ 800 nM) of Pb²⁺ in real water samples (ground water and tap water). Specially, 2 was successfully applied for the quantification of Pb²⁺ in human serum by combination of microwave-assisted human serum digestion and filtration of digested serum by anion exchange cartridge. We clearly investigated the binding mode of 2 with Pb²⁺ using ¹H NMR, IR spectroscopy, pH titration, confocal microscopy, and size analysis. The peptide-based fluorescent probe might have great application potential for sensing Pb²⁺ in aquatic environments and biofluid samples.