A pyrazolopyrimidine based fluorescent probe for the detection of Cu2+ and Ni2+ and its application in living cells

Abstract

A fluorescent sensor L based on a pyrazolopyrimidine core simultaneously detects Cu²⁺ and Ni²⁺ ions by photoluminescence quenching, even in the presence of other metal cations. Sensor L possesses high association constants of 5.24 × 10³ M^-1 and 2.85 × 10⁴ M^-1 and low detection limits of 0.043 μM and 0.038 μM for Cu²⁺ and Ni²⁺, respectively. The binding stoichiometry ratios of L to Cu²⁺ or Ni²⁺ is 1:1 as determined by Benesi-Hildebrand and Job's plots, and by crystal structures. DFT calculations on L-Cu²⁺ indicated reduced electron donation from the coordinated pyrazolopyridine to the fused pyrimidine and pendant phenyl group which, together with a smaller HOMO-LUMO orbital gap could favour non-radiative decay and explain the observed fluorescence quenching. Sensor L possessed low cytotoxicity and good imaging characteristics for Cu²⁺ and Ni²⁺ in living cells, suggesting potential applications for detecting Cu²⁺ and Ni²⁺in vivo.

Keywords: Cu(2+) sensing, Ni(2+) sensing; Fluorescent sensor; Pyrazolopyrimidines.