A novel label-free fluorescence assay for one-step sensitive detection of Hg2+ in environmental drinking water samples

Abstract

A novel label-free fluorescence assay for detection of Hg²⁺ was developed based on the Hg²⁺-binding single-stranded DNA (ssDNA) and SYBR Green I (SG I). Differences from other assays, the designed rich-thymine (T) ssDNA probe without fluorescent labelling can be rapidly formed a T-Hg²⁺-T complex and folded into a stable hairpin structure in the presence of Hg²⁺ in environmental drinking water samples by facilitating fluorescence increase through intercalating with SG I in one-step. In the assay, the fluorescence signal can be directly obtained without additional incubation within 1 min. The dynamic quantitative working ranges was 5-1000 nM, the determination coefficients were satisfied by optimization of the reaction conditions. The lowest detection limit of Hg²⁺ was 3 nM which is well below the standard of U.S. Environmental Protection Agency. This method was highly specific for detecting of Hg²⁺ without being affected by other possible interfering ions from different background compositions of water samples. The recoveries of Hg²⁺ spiked in these samples were 95.05-103.51%. The proposed method is more viable, low-costing and simple for operation in field detection than the other methods with great potentials, such as emergency disposal, environmental monitoring, surveillance and supporting of ecological risk assessment and management.