title: Ellman's and Aldrithiol Assay as Versatile and Complementary Tools for the Quantification of Thiol Groups and Ligands on Nanomaterials.
abstract: Simple, fast, and versatile methods for the quantification of thiol groups are of considerable interest not only for protein analysis but also for the characterization of the surface chemistry of nanomaterials stabilized with thiol ligands or bearing thiol groups for the subsequent (bio-) functionalization via maleimide-thiol chemistry. Here, we compare two simple colorimetric assays, the widely used Ellman's assay performed at alkaline pH and the aldrithiol assay executed at acidic and neutral pH, with respect to their potential for the quantification of thiol groups and thiol ligands on different types of nanoparticles like polystyrene nanoparticles, semiconductor nanocrystals (SC NC), and noble metal particles, and we derive criteria for their use. In order to assess the underlying reaction mechanisms and to obtain stoichiometry factors mandatory for reliable thiol quantification, both methods were studied photometrically and with electrospray ionization time-of-flight mass spectrometry (ESI-TOF-MS), thereby demonstrating the influence of different thiols on the reaction mechanism. Our results underline the suitability of both methods for the quantification of directly accessible thiol groups or ligands on the surface of 2D- and 3D-supports, here exemplarily polystyrene nanoparticles. Moreover, we could derive strategies for the use of these simple assays for the determination of masked (i.e., not directly accessible) thiol groups like disulfides such as lipoic acid and thiol stabilizing ligands coordinatively bound to Cd and/or Hg surface atoms of II/VI and ternary SC NC and to gold and silver nanoparticles.
authors.affiliation: Federal Institute for Materials Research and Testing (BAM), Richard-Willstaetter-Str. 11, D-12489 Berlin, Germany.
authors.forename: Marko
authors.initials: M
authors.lastname: Moser
authors.affiliation: Federal Institute for Materials Research and Testing (BAM), Richard-Willstaetter-Str. 11, D-12489 Berlin, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Schneider
authors.affiliation: Federal Institute for Materials Research and Testing (BAM), Richard-Willstaetter-Str. 11, D-12489 Berlin, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Behnke
authors.affiliation: Federal Institute for Materials Research and Testing (BAM), Richard-Willstaetter-Str. 11, D-12489 Berlin, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Schneider
authors.affiliation: Federal Institute for Materials Research and Testing (BAM), Richard-Willstaetter-Str. 11, D-12489 Berlin, Germany.
authors.forename: Jana
authors.initials: J
authors.lastname: Falkenhagen
authors.affiliation: Federal Institute for Materials Research and Testing (BAM), Richard-Willstaetter-Str. 11, D-12489 Berlin, Germany.
authors.forename: Ute
authors.initials: U
authors.lastname: Resch-Genger