• collision-induced dissociation;
  • electrospray ionization;
  • flavonoid C-glycoconjugates and/or O-glycoconjugates;
  • liquid chromatography/mass spectrometry;
  • secondary metabolite profiling;
  • structural characterization;
  • wheat

The aim of this study was to present integrated mass spectrometric methods for the structural characterization and identification of flavonoid glycoconjugates. During the liquid chromatography/mass spectrometry analyses, TriVersa NanoMate chip-based system with nanoelectrospray ionization and fraction collection was combined to a quadrupole time-of-flight mass spectrometer. In the extract samples prepared from green leaves of wheat plantlets, 41 flavonoid derivatives were recognized. Part of the target natural products had the full structure being characterized after the registration of mass spectra, where m/z values for protonated [M + H]+ and deprotonated molecules [M − H] were annotated. MS2 and pseudo-MS3 experiments were performed for [M + H]+ or [M − H] and aglycone ions (Y0+/−-type), respectively. It should be underlined that pseudo-MS3 mass spectra were registered for aglycone product ions in the mass spectra of O-glycosides present in the extract samples. In many cases, only tentative structural identification of aglycones was possible, mainly because of the presence of numerous C-monoglycoside or C-diglycoside in the samples. Acylation of the sugar moiety and/or methylation of the aglycone in the flavonoid glycosides under study was observed. The existence of isobaric and/or isomeric compounds was demonstrated in the extract studied. The collision-induced dissociation mass spectra registered for C,O-diglycosides and C,C-diglycosides did not permit to draw complete structural conclusions about the compounds studied. For the investigated class of natural products, unambiguous classification of sugar moieties linked to the aglycones from the recorded mass spectra was not possible. Registration of the positive and negative ion mass spectra did not lead to the precise conclusion about the glycosylation position at C-6 or C-8, and O-4′ or O-7 atoms. It was possible, on the basis of the collected MS2 spectra, to differentiate between O-glycosides and C-glycosides present in the samples analyzed. Copyright © 2013 John Wiley & Sons, Ltd.