Platform- and label-free detection of lead ions in environmental and laboratory samples using G-quadraplex probes by circular dichroism spectroscopy

Guanine-rich quadruplex (G-QD) are formed by conversion of nucleotides with specific sequences by stabilization of positively charged K⁺ or Na⁺. These G-QD structures differentially absorb two-directional (right- and left-handed) circularly polarized light, which can discriminate the parallel or anti-parallel structures of G-QDs. In this study, G-QDs stabilized by Pb²⁺ were analyzed by a circular dichroism (CD) spectroscopy to determine Pb²⁺ concentration in water samples. Thrombin aptamer (TBA), PS2.M, human telomeric DNA (HTG), AGRO 100, and telomeric related sequence (T2) were studied to verify their applicability as probes for platform- and label-free detection of Pb²⁺ in environmental as well as laboratory samples. Among these nucleotides, TBA and PS2.M exhibited higher binding constants for Pb²⁺, 1.20–2.04 × 10⁶/M at and 4.58 × 10⁴–1.09 × 10⁵/M at 100 micromolar and 100 mM K⁺ concentration, respectively. They also exhibited excellent selectivity for Pb²⁺ than for Al³⁺, Cu²⁺, Ni²⁺, Fe³⁺, Co²⁺, and Cr²⁺. When Pb²⁺ was spiked into an effluent sample from a wastewater treatment plant (WWTP), its existence was detected by CD spectroscopy following a simple addition of TBA or PS2.M. By the addition of TBA and PS2.M, the Pb²⁺ signals were observed in effluent samples over 0.5 micromolar (100 ppb) concentration. Furthermore, PS2.M caused a Pb²⁺-specific absorption band in the effluent sample without spiking of Pb²⁺, and could be induced to G-QD structure by the background Pb²⁺ concentration in the effluent, 0.159 micromolar concentration (3.30 ppb). Taken together, we propose that TBA and PS2.M are applicable as platform- and label-free detection probes for monitoring Pb²⁺ in environmental samples such as discharged effluent from local WWTPs, using CD spectroscopy.