Abstract
Profiling of plant secondary metabolites is still a very difficult task. Liquid chromatography (LC) or capillary electrophoresis hyphenated with different kinds of detectors are methods of choice for analysis of polar, thermo labile compounds with high molecular masses. We demonstrate the applicability of LC combined with UV diode array or/and mass spectrometric detectors for the unambiguous identification and quantification of flavonoid conjugates isolated from Arabidopsis thaliana leaves of different genotypes and grown in different environmental conditions. During LC/UV/MS/MS analyses we were able to identify tetra-, tri-, and di-glycosides of kaempferol, quercetin and isorhamnetin. Based on our results we can conclude that due to the co-elution of different chemical compounds in reversed phase HPLC systems the application of UV detectors does not allow to precisely profile all flavonoid conjugates existing in A. thaliana genotypes. Using MS detection it was possible to unambiguously recognize the glycosylation patterns of the aglycones. However, from the mass spectra we could not conclude neither the anomeric form of the C-1 carbon atoms of sugar moieties in glycosidic bonds between sugars or sugar and aglycone nor the position of the second carbon involved in disaccharides. The applicability of collision induced dissociation techniques (CID MS/MS) for structural analyses of the studied group of plant secondary metabolites with two types of analyzers (triple quadrupole or ion trap) was demonstrated.
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Acknowledgments
We greatly thank Matthew Hannah for improving our English, and Dr. Karin Koehl and her colleagues from the Green Team for expert plant care. Both are at the MPI of Molecular Plant Physiology, Golm.
Acknowledgement of financial support: Aleksandra Skirycz gratefully acknowledges the Ernst Schering Foundation, Berlin, for providing a doctoral fellowship. Additional financial support was provided through the International PhD Programme ’Integrative Plant Science’ (IPP-IPS) funded by the DAAD (Deutscher Akademischer Austauschdienst) and the DFG (Deutsche Forschungsgemeinschaft) under No. DAAD Az. D/04/01336, by the Interdisciplinary Research Centre ’Advanced Protein Technologies’ (IZ-APT) of the University of Potsdam, and by the Fonds der Chemischen Industrie (No. 0164389).
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Stobiecki, M., Skirycz, A., Kerhoas, L. et al. Profiling of phenolic glycosidic conjugates in leaves of Arabidopsis thaliana using LC/MS. Metabolomics 2, 197–219 (2006). https://doi.org/10.1007/s11306-006-0031-5
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DOI: https://doi.org/10.1007/s11306-006-0031-5