Background: Nanozymes have the advantages of cost effective, simple synthesis, high durability and stability, and have been widely used in various fields. However, only a few nanomaterials with multiple enzyme-like activity have been reported, and most of the currently developed nanozymes are usually used in colorimetric or fluorescence analysis depending on a single colorimetric or fluorescence signal output. In this study, a copper-based dual-ligand biomimetic nanoflower (Cu-MN) was constructed, which demonstrated potential multiple enzyme-like activity, and was applied to the multi-mode detection of 2,4-dichlorophenol (2,4-DP), epinephrine (EP), and H₂O₂.

Results: The laccase-like activity of Cu-MN can catalyze the conversion of 2,4-DP and EP, resulting in the formation of red and yellow-brown oxidation products with distinct UV absorption peaks at 510 nm and 485 nm, respectively. Furthermore, the fluorescence emission peak at 426 nm of Cu-MN can be dynamic quenched during substrate oxidation due to the fluorescence internal filtration effect (IFE). Therefore, a dual-mode analysis method was constructed to detect 2,4-DP and EP by fluorescence and ultraviolet colorimetry, which was successfully applied in natural lake water and rabbit plasma analysis, respectively. Furthermore, a colorimetric sensing strategy based on the peroxidase-like activity of Cu-MN was developed and successfully applied to the monitoring of H₂O₂ in hydrogen peroxide disinfectant. Additionally, the visualization analysis method was also established by RGB reading of the smartphone.

Significance and novelty: In brief, inspired by the fluorescence characteristics of 2-aminoterephthallc acid and the imidazole group of 2-methylimidazole, a novel copper-based dual-ligand biomimetic nanoflower (Cu-MN) was prepared and used to establish multi-mode method for the detection of 2,4-DP, EP, and H₂O₂, which opens up new avenues for its applications in bioanalysis and environmental monitoring.