Simultaneous colorimetric and electrochemical detection of trace mercury (Hg2+) using a portable and miniaturized aptasensor

Abstract

We report a novel aptasensor for the simultaneous colorimetric and electrochemical detection of mercury (Hg²⁺). This device consists of a paper-based microfluidic component (μ-PAD) incorporated into a miniaturized three-electrode system fabricated through printed circuit board (PCB) technology. This biosensor is portable, rapid, versatile, and can detect Hg²⁺ down to 0.01 ppm based on 3σ of the blank/slope criteria. Moreover, it is highly selective against As²⁺, Cu²⁺, Zn²⁺, Pb²⁺, Cd²⁺, Mg²⁺, and Fe²⁺, reaching up to 13 times more of the input signal than the other heavy metals. The colorimetric detection mechanism uses aptamer functionalized polystyrene (PS)-AgNPs and Ps-AuNPs microparticles' specific aggregation. The Ps-AuNPs-based system allows qualitative detection (LOD 5 ppm) and stability over seven days (up to 97.59% signal retention). For the Ps-AgNPs-based system, the detection limit is 0.5 ppm with a linear range from 0.5 to 20 ppm (adjusted R²= 0.986) and stability over 30 days (up to 94.95% signal retention). The electrochemical component measures changes in charge transfer resistance upon target-aptamer hybridization using a [Ru (NH₃)₆]³⁺Cl₃] redox probe. The latest component presents a linear range from 0.01 to 1 ppm (adjusted R²= 0.935) with a LOD of 0.01 ppm and performance stability over seven days (up to 102.52 ± 11.7 signal retention). This device offers a universal dual detection platform with multiplexing, multi-replication, quantitative color analysis, and minimization of false results. Furthermore, detection results in river samples showed recoveries up to 91.12% (RSD 0.85) and 105.61% (RSD 1.62) for the electrochemical and colorimetric components, respectively. The proposed system is highly selective with no false-positive or false-negative results in an overall wide linear range and can safeguard the accuracy of detection results in aptasensing platforms in general.

Keywords: Colorimetric; Dual-modality; Electrochemical; Multiplexing; On-site.