title: Data-directed scan sequence for the general assignment of C-glycosylflavone O-glycosides in plant extracts by liquid chromatography-ion trap mass spectrometry.
abstract: An ion trap LC-MS/MS method is described for the analysis of C-glycosylflavone O-glycosides in crude methanolic extracts of plants. The method employs survey scans with and without the application of up-front collision induced dissociation (CID) to generate diagnostic ions for data-directed MS/MS. The spectra acquired allow assignment of the C-linked sugar to either the C-6 or C-8 position of the aglycone and provide data on the molecular mass of the compound, the number and type of O-linked sugars and the molecular mass of the flavone aglycone. These data for the majority of C-glycosylflavone O-glycosides in an extract are obtained automatically in one LC-MS/MS analysis without manual pre-programming. Key to the assignment of the C-6 or C-8 site of C-glycosylation is the generation, by up-front CID, of the (0,1)X+ product ion formed by internal cleavage of the C-linked sugar. MS/MS of this ion is found to have diagnostic value in addition to the (0,2)X+ product ion described by other authors. Ion trap MS/MS spectra of [M+H]+ of the 6,8-di-C-glycosylflavones schaftoside and isoschaftoside show an additional and previously unreported diagnostic product ion that is useful in determining the type of sugar at the C-6 position. The product ion spectra of protonated kaempferol 3-O-glucosylrhamnosides show similarities to the spectra of C-glycosylflavone O-glycosides; this is a potential source of confusion if the analysis of such glycosides is limited solely to MS/MS of [M+H]+.
authors.affiliation: Royal Botanic Gardens, Kew, Richmond, Surrey TW9 3AB, UK. g.kite@kew.org
authors.forename: Geoffrey C
authors.initials: GC
authors.lastname: Kite
authors.forename: Elaine A
authors.initials: EA
authors.lastname: Porter
authors.forename: Fiona C
authors.initials: FC
authors.lastname: Denison
authors.forename: Renée J
authors.initials: RJ
authors.lastname: Grayer
authors.forename: Nigel C
authors.initials: NC
authors.lastname: Veitch
authors.forename: Ian
authors.initials: I
authors.lastname: Butler
authors.forename: Monique S J
authors.initials: MS
authors.lastname: Simmonds
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D047309:Flavones / Q000737:chemistry*; D006027:Glycosides / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry / Q000379:methods*; D010936:Plant Extracts / Q000737:chemistry*
chemical_list: D047309:Flavones; D006027:Glycosides; D010936:Plant Extracts