In the work the focus is on the preparation of self-assembled monolayer-like films consisting of thiolated cyclodextrin on gold substrate and a characterization by using secondary ion mass spectrometry. The short (1 min) and long (1 h) time preparations of self-assembled monolayer-like films, resulting in submonolayer and monolayer regimes, are investigated, respectively. The observed species of thiolated cyclodextrin (M as molecular ion) self-assembled monolayer-like films are assigned to three groups: AuxHySz clusters, fragments with origin in cyclodextrin molecule associated with Au, and molecular ions. The group of AuxHySz (x = 2-17, y = 0-2, z = 1-5) clusters have higher intensities than other species in the positive and even more in negative mass spectra. Interestingly, the dependence between the number of Au and S atoms shows that with the increasing size of AuxHySz clusters up to 11 Au atoms, the number of associated S atoms is also increasing and then decreasing. Molecular species as (M-S+H)Na+, (M+H)Na+, AuMNa+, (M-2-S) Na+, and M2Na+ are determined, and also in cationized forms with K+. The intensities of thiolated cyclodextrin fragments at the long time preparation are approximately 10 times higher than the intensities of the same fragments observed at the short time. The largest observed ions in thiolated cyclodextrin self-assembled monolayer-like films are AuM2 and Au2M. The thiolated cyclodextrin molecular ions are compared with hexadecanethiol molecular ions in the form of AuxMw where the values of x and w are smaller for thiolated cyclodextrin than for hexadecanethiol. This result is supported with larger, more compact, and more stabile thiolated cyclodextrin molecule. (C) 2010 Elsevier B.V. All rights reserved.