Colorimetric determination of p-nitrophenol by using ELISA microwells modified with an adhesive polydopamine nanofilm...

A microplate method is described for the quantification of p-nitrophenol (p-NPh) in urine samples where it can be found after exposure to certain insecticides such as methyl parathion or paraoxon. The assay is based on the use of a polydopamine (PDA) film doped with gold nanoparticles (AuNPs). The latter exerts a catalytic effect on the reduction of nitrophenols by NaBH₄. PDA has adhesive properties and can be used to fix the AuNPs on several solid substrates, here ELISA polystyrene microwells. The optical and catalytic properties of different populations of AuNPs spontaneously grown on PDA films were investigated, mainly in terms of the relationship between AuNPs@PDA nanocomposite preparation and its catalytic activity and stability. The reduction of o-, m-, and p-nitrophenols by NaBH₄ in aqueous solution was exploited as model study. The approach demonstrates that useful kinetic information on the catalytic effect can be obtained on 96-wells simultaneously by a conventional ELISA reader at a fixed wavelength of 415 nm. The method was successfully applied to the quantification of p-NPh in (spiked) urine samples and gave high reproducibility (RSD = 3.5%) and a 6.30 μM (836 μg/L) detection limit. Graphical abstract Schematic presentation of 96-wells preparation for optical quantification on ELISA reader of p-nitrophenol (p-NPh) catalytic reduction to p-aminophenol (p-APh), as model study, by NaBH₄ and different population gold nanoparticles (AuNPs) grown on polydopamine (PDA) films attached onto polystyrene (PS) wells.