A methionine biomolecule-modified chromenylium-cyanine fluorescent probe for the analysis of Hg²⁺ in the environment and living cells

The objective of the study is, for the first time, to construct a new near infrared (NIR) fluorophore, spectrophotometric, colorimetric, ratiometric, and turn-on probe (CSME) based on chromenylium cyanine platform decorated with methionine biomolecule to provide an efficient solution for critical shortcoming to be encountered for analysis of hazardous Hg²⁺ in environment and living cell. The CSME structure and its interaction with Hg²⁺ ion were evaluated by NMR, FTIR, MS, UV-Vis and fluorescence methods as well as Density Functional Theory (DFT) calculations. The none fluorescence CSME having spirolactam ring only interacted with Hg²⁺ in aqueous solution including competing ions. This interaction caused the fluorescence CSME with opened spirolactam form which exhibited spectral and colorimetric changes in the NIR region. The probe based on UV-Vis and fluorescence techniques respond in 90 s, has wide linear ranges (for UV-Vis: 6.29 × 10^-8 - 1.86 × 10^-4 M; for fluorescence: 9.49 × 10^-9 - 1.13 × 10^-5 M), and has a lower Limit of Detection (LOD) value (for fluorescence: 4.93 × 10^-9 M, 0.99 ng/mL) than the value predicted by the US Environmental Protection Agency (EPA) organization. Hg²⁺ analysis was performed in drinking and tap water with low Relative Standard Deviation (RSD) values and high recovery. Smartphone and living cell applications were successfully performed for colorimetric sensing Hg²⁺ in real samples and 3T3 cells, respectively.