title: Clarification of pathway-specific inhibition by Fourier transform ion cyclotron resonance/mass spectrometry-based metabolic phenotyping studies.
abstract: We have developed a metabolic profiling scheme based on direct-infusion Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS). The scheme consists of: (1) reproducible data collection under optimized FT-ICR/MS analytical conditions; (2) automatic mass-error correction and multivariate analyses for metabolome characterization using a newly developed metabolomics tool (DMASS software); (3) identification of marker metabolite candidates by searching a species-metabolite relationship database, KNApSAcK; and (4) structural analyses by an MS/MS method. The scheme was applied to metabolic phenotyping of Arabidopsis (Arabidopsis thaliana) seedlings treated with different herbicidal chemical classes for pathway-specific inhibitions. Arabidopsis extracts were directly infused into an electrospray ionization source on an FT-ICR/MS system. Acquired metabolomics data were comprised of mass-to-charge ratio values with ion intensity information subjected to principal component analysis, and metabolic phenotypes from the herbicide treatments were clearly differentiated from those of the herbicide-free treatment. From each herbicide treatment, candidate metabolites representing such metabolic phenotypes were found through the KNApSAcK database search. The database search and MS/MS analyses suggested dose-dependent accumulation patterns of specific metabolites including several flavonoid glycosides. The metabolic phenotyping scheme on the basis of FT-ICR/MS coupled with the DMASS program is discussed as a general tool for high throughput metabolic phenotyping studies.
authors.affiliation: Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai 599-8531, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Oikawa
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Tomonori
authors.initials: T
authors.lastname: Ogura
authors.forename: Atsuko
authors.initials: A
authors.lastname: Kimura
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Suzuki
authors.forename: Nozomu
authors.initials: N
authors.lastname: Sakurai
authors.forename: Yoko
authors.initials: Y
authors.lastname: Shinbo
authors.forename: Daisuke
authors.initials: D
authors.lastname: Shibata
authors.forename: Shigehiko
authors.initials: S
authors.lastname: Kanaya
authors.forename: Daisaku
authors.initials: D
authors.lastname: Ohta
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D017357:Cyclotrons; D005583:Fourier Analysis; D018525:Germination; D006540:Herbicides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D013058:Mass Spectrometry / Q000379:methods*; D015394:Molecular Structure; D010641:Phenotype; D036226:Seedlings / Q000378:metabolism
chemical_list: D006540:Herbicides