Plasmonic active film integrating gold/silver nanostructures for H₂O₂ readout

A nanostructured Ag/Au adhesive film for H₂O₂ reagentless determination is here proposed. The film has been realised onto ELISA polystyrene microplates. Microwells surface has been initially modified with a gold nanoparticles (AuNPs)/polydopamine thin-film. The pristine AuNPs-decorated film was later functionalized with catechin (Au-CT) allowing a uniform formation of a plasmonic active nanostructured silver network in presence of Ag⁺. Changes in localized surface plasmon resonance (LSPR) of the silver network upon addition of H₂O₂ has been used as analytical signal, taking advantage of the etching phenomenon. The Ag/Au nanocomposite-film is characterized by a well-defined (LSPR_max = 405 ± 5 nm), reproducible (intraplate RSD ≤ 9.8%, n = 96; inter-plate RSD ≤ 11.4%, n = 480) and stable LSPR signal. The film's analytical features have been tested for H₂O₂ and glucose (bio)sensing. Satisfactory analytical performances were obtained both for H₂O₂ (linear range 1-200 μM, R² = 0.9992, RSD ≤ 6.3%, LOD = 0.2 μM) and glucose (linear range 2-250 μM, R² = 0.9998, RSD ≤ 8.9%, LOD = 0.4 μM). As proof of applicability, the determination of the two analytes in soft drinks has been carried out achieving good and reproducible recoveries (84-111%; RSD ≤ 9%). The developed nanostructured film overcomes analytical drawbacks associated with the use of colloidal dispersions in plasmonic assays carried out in solution; the low cost, robustness, ease of use and possibility of coupling enzymatic reactions appears very promising for (bio)sensors based on the detection of H₂O₂.