Development of a Ratiometric Fluorescent Cu(II) Indicator Based on Poly(N-isopropylacrylamide) Thermal Phase Transition and an Aminopyridyl Cu(II) Ligand

An aqueous Cu2+ and Zn2+ indicator is reported based on copolymerizing aminopyridine ligands and the environment-sensitive dansyl fluorophore into the responsive polymer poly(N-isopropylacrylamide) (PNIPAm). The metal ion binding creates charge and solvation that triggers PNIPAm’s thermal phase transition from hydrophobic globule to hydrophilic open coil. As a basis for sensing the metal-binding, the dansyl fluorescence emission spectra provide a signal at ca. 530 nm and a signal at 500 nm for the hydrophobic and hydrophilic environment, respectively, that are ratiometrically interpreted. The synthesis of the title pyridylethyl-pyridylmethyl-amine ligand (acronym PEPMA) with a 3-carbon linker to the copolymerizable group, aminopropylacrylamide (PEPMA-C3-acrylamide), is reported, along with a nonpolymerizable model ligand derivative. The response of the polymer is validated by increasing temperature from 25 °C to 49 °C, which causes a shift in maximum emission wavelength from 536 nm to 505 nm, along with an increase in the ratio of emission intensity of 505 nm/536 nm from 0.77 to 1.22 (λex = 330 nm) as the polymer releases water. The addition of divalent Cu or Zn to the indicator resulted in a dansyl emission shift of 10 nm to a longer wavelength, accompanied by fluorescence quenching in the case of Cu2+. The addition of EDTA to the Cu2+-loaded indicator reversed the fluorescence shift at 25 °C to 35 °C. The affinities of Cu2+ and Zn2+ for the PEPMA derivatives are log Kf = 11.85 and log Kf = 5.67, respectively, as determined by potentiometric titration. The single-crystal X-ray structure of the Cu2+-PEPMA derivative is five-coordinate, of-geometry intermediate between square-pyramidal and trigonal-bipyramidal, and is comparable to that of Cu2+ complexes with similar formation constants.