Microchimica Acta

, 186:146 | Cite as

Colorimetric determination of p-nitrophenol by using ELISA microwells modified with an adhesive polydopamine nanofilm containing catalytically active gold nanoparticles

  • Simona ScaranoEmail author
  • Pasquale Palladino
  • Emanuela Pascale
  • Alvaro Brittoli
  • Maria Minunni
Original Paper


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.


Nanocomposite materials Urine analysis Metallic nanostructures Biopolymer Redox catalyst High-throughput screening UV-vis spectroscopy Plasmons Catechols 



Authors thank the Ministry of Education, University and Research (MIUR) for financial support through the scientific program SIR2014 Scientific Independence of young Researchers (RBSI1455LK).

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2019_3259_MOESM1_ESM.docx (2.3 mb)
ESM 1 (DOCX 2.27 mb)


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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Simona Scarano
    • 1
    Email author
  • Pasquale Palladino
    • 1
  • Emanuela Pascale
    • 1
  • Alvaro Brittoli
    • 1
  • Maria Minunni
    • 1
  1. 1.Department of Chemistry ‘Ugo Schiff’University of FlorenceSesto FiorentinoItaly

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