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Updated: Aug 7, 2026
Sample Preparation for Mass Spectrometry-based Identification of RNA-binding Regions
Published on: September 28, 2017
Mass spectrometric identification and characterization of antimony complexes with ribose-containing biomolecules and
Helle Rüsz Hansen1, Spiros A Pergantis
1Department of Chemistry, Environmental Chemical Processes Laboratory, University of Crete, P.O. Box 2208, Voutes, 71003 Heraklion, Greece.
Abstract:
Mass spectrometric techniques have been used to study the interaction of inorganic Sb(V) with biomolecules containing a ribose or deoxyribose moiety. Electrospray (ES) mass spectra of reaction mixtures containing inorganic Sb(V) and one of several biomolecules (adenosine, cytidine, guanosine, uridine, adenosine-5'-monophosphate, adenosine-3',5'-cyclic monophosphate, ribose, or 2'-deoxyadenosine) afforded high-mass antimony-containing ions corresponding to Sb(V)-biomolecule complexes of stoichiometry 1:1, 1:2, or 1:3. The complexes were characterized by collision-induced dissociation (CID) tandem mass spectrometry (MS) using ion-trap multistage MS. The CID results revealed that Sb(V) binds to the ribose or deoxyribose moiety. Structures are proposed for the Sb-biomolecule complexes. Analysis of the reaction mixtures by reversed-phase chromatography coupled on-line to either inductively coupled plasma (ICP) MS or ES-MS showed that in solution Sb(V) forms complexes with all the analyzed biomolecules with vicinal cis hydroxyl groups. Evidence (from size-exclusion chromatography ICP-MS and direct infusion ES-MS) of complexation of Sb(V) with an RNA oligomer, but not with a DNA oligomer, supports the suggestion that the presence of vicinal cis hydroxyl groups is critical for complexation to occur. This is the first direct evidence of complexation of Sb(V) with RNA. Results obtained by studying the effect of changing reaction conditions, i.e. pH, reaction time, and Sb/biomolecule molar ratio, on the extent of Sb-biomolecule formation suggest the reaction may be of physiological importance. Selected reaction monitoring (SRM) and precursor-ion-scanning tandem MS were investigated to determine their potential to detect trace levels of the Sb-biomolecule complexes in biological samples. Application of SRM MS-MS in combination with high-performance liquid chromatography enabled successful detection of an Sb-adenosine complex that had been spiked into a complex biological matrix (liver homogenate).
