Pitfalls and capabilities of various hydrogen donors in evaluation of peroxidase-like activity of gold nanoparticles

Marcin Drozd , Mariusz Pietrzak , Paweł G. Parzuchowski , Elżbieta Malinowska


Catalytic nanomaterials, widely used as substitutes of peroxidase, exhibit unique properties, which are unattainable for native enzymes. However, their activity is usually examined by means of substrates developed and methods standardized for horseradish peroxidase (HRP). The aim of the presented work was to determine the scope of usefulness of chromogenic substrates for gold nanoparticle (AuNP) activity studies under conditions which significantly extend beyond the activity range of a native HRP. The applicability of chromogens such as 3,3′5,5′-tetramethylbenzidine (TMB), o-phenylenediamine (OPD), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) beyond the typical range of pH, and for the samples of high concentration of hydrogen peroxide was examined. The conducted research confirmed the usefulness of ABTS and TMB in acidic media (pH 2.5–3.5). At the same time, potential interferences from chloride anion, unobservable for HRP-based assays, were indicated. Moreover, a number of potentially useful hints concerning relations of concentration of substrates and catalyst for aromatic amine oxidation (TMB and OPD) were proposed. By increasing the concentration of chromogens and thanks to assuring the relatively low conversion of the reaction, the stability of TMB and OPD oxidation product was improved even in acidic media. The comparative studies of H2O2 affinity to the surface of AuNPs in the presence of various hydrogen donors underlined the superiority of phenolic compounds over aromatic amines and ABTS in the case of the samples of relatively low H2O2 concentration. This work highlights some improvements in the methods of HRP-like activity characterization of NPs. It provides a critical analysis of the major challenges, which may emerge in a case of bioanalytical assays employing the catalytic nanoparticles as labels.
Author Marcin Drozd ZMB
Marcin Drozd,,
- Department Of Microbioanalytics
, Mariusz Pietrzak ZMB
Mariusz Pietrzak,,
- Department Of Microbioanalytics
, Paweł G. Parzuchowski KChTP
Paweł G. Parzuchowski,,
- Chair Of Polymer Chemistry And Technology
, Elżbieta Malinowska ZMB
Elżbieta Malinowska,,
- Department Of Microbioanalytics
Journal seriesAnalytical and Bioanalytical Chemistry, ISSN 1618-2642 [1618-2650, 0937-0633]
Issue year2016
Publication size in sheets0.7
Keywords in EnglishCatalytic activity; Gold nanoparticles; Peroxidase mimetics; Nanozyme
URL http://link.springer.com/article/10.1007/s00216-016-9976-z
Languageen angielski
art%3A10.1007%2Fs00216-016-9976-z.pdf (file archived - login or check accessibility on faculty) art%3A10.1007%2Fs00216-016-9976-z.pdf 1.62 MB
Score (nominal)40
ScoreMinisterial score = 35.0, 28-11-2017, ArticleFromJournal
Ministerial score (2013-2016) = 40.0, 28-11-2017, ArticleFromJournal
Publication indicators WoS Impact Factor: 2016 = 3.431 (2) - 2016=3.306 (5)
Citation count*0
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.