Copper-based fluorescent nanozyme used to construct a ratiometric sensor for visual detection of thiophanate methyl.

Although nanozyme has shown great potential in designing fluorescent assays for pesticide residue, most of them are based on single emission, thus affecting the detection accuracy. Herein, a copper-based fluorescent nanozyme (Cu-BH) synthesized with dual-ligand, integrating fluorescence and oxidase-mimic into one spherical nanomaterial, was used firstly to establish a ratiometric approach for visual detection of thiophanate methyl (TM). Cu-BH possesses excellent oxidase-like activities, triggering the oxidation of colorless o-phenylenediamine (OPD) into yellow luminescent products (oxOPD, λem = 564 nm). Besides, the ligand of 2-amino-1,4-benzene-dicarboxylic acid imparts Cu-BH blue fluorescence (λem = 425 nm), which is quenched by oxOPD via inner filtration effect (IFE). The introduction of TM can prevent not only the oxidase-like activity remarkably but also the intrinsic luminescence of Cu-BH slightly because of the complexation of TM with Cu²⁺. As a result, the fluorescence intensity at 564 nm and 425 nm presents a significant decrease and a slight increase, respectively, producing a ratiometric fluorescent signal (F₄₂₅/F₅₆₄). Therefore, a novel ratiometric fluorescent strategy has been proposed to detect TM ranging from 0.1 to 100 μM with detection limit of 0.03 μM (S/N = 3). Besides, visual detection of TM can be achieved by RGB reading with the assistance of smartphone owing to the color variation from yellow to blue. This fluorescent nanozyme-based ratiometric strategy provides a specific method for the detection of TM in food samples.