Ratiometric fluorescent detection of lead ions in aquatic environment and living cells using a fluorescent peptide-based probe.

Ratiometric fluorescent detection using dual emission bands is highly necessary to quantify Pb(II) in aquatic environment and live cells. We synthesized a ratiometric fluorescent peptidyl probe (1) by conjugation of a peptide receptor for Pb(II) with an excimer-forming benzothiazolylcyanovinylene fluorophore. The peptidyl probe dissolved well in aqueous solution and displayed an emission band at 538 nm (λex = 460 nm). Upon addition of Pb(II) (0-20 µM), the emission maximum shifted from 538 nm to 575 nm and the emission intensity ratio (I575 /I538) increased significantly from 0.40 to 2.26. 1 exhibited a selective ratiometric response to Pb(II) over other metal ions. 1 with a low detection limit (1.2 ppb) of Pb(II) detected nanomolar concentrations (0-500 nM) of Pb(II) ions in groundwater and tap water. The cell-permeable probe detected intracellular Pb(II) by ratiometric fluorescent images. The binding mode study using NMR, IR and CD spectroscopy, and TEM revealed that the probe formed a 1:1 complex with Pb(II) and then formed red-emissive nanoparticles and fibrils. The probe exhibited desirable detection properties such as ratiometric detection, high solubility in water, visible light excitation, high selectivity and sensitivity for Pb(II), cell-permeability, and rapid response (< 6 min).

Fluorescent Detection of Methyl Mercury in Aqueous Solution and Live Cells Using Fluorescent Probe and Micelle Systems.

It is highly challenging to develop fast and sensitive fluorescent methods for monitoring organic mercury in purely aqueous solutions as well as live cells. Especially, selective fluorescent detection of methylmercury over inorganic mercury ions has not been reported. We developed a fast and sensitive fluorescent detection method for Hg2+ ions as well as methylmercury using an amino acid-based fluorescent probe (1) and SDS micelles. The fluorescent probe in SDS micelles detected sensitively and selectively Hg2+ ions and methylmercury among 16 metal ions in purely aqueous solution by the enhancement of the red emission at 575 nm, and the detection of methylmercury was completed within 1 min. The probe in SDS micelles with EDTA showed highly sensitive and selective turn on detection for methylmercury over Hg2+. The limit of detection was 9.1 nM for Hg2+ (1.8 ppb, R2 = 0.989) and 206 nM for CH3Hg+ (R2 = 0.997). 1 rapidly penetrated live cells and detected intracellular Hg2+ ions as well as CH3Hg+ by the enhancement of both red emissions and green emissions. Subsequent treatment of EDTA into the cell confirmed the selective detection of methylmercury in the cells. The present work indicated that the fluorescent probe with micelle systems provided a fast, sensitive, and selective detection method for monitoring inorganic mercury as well as methyl mercury.