title: Stable isotope assisted assignment of elemental compositions for metabolomics.
abstract: Assignment of individual compound identities within mixtures of thousands of metabolites in biological extracts is a major challenge for metabolomic technology. Mass spectrometry offers high sensitivity over a large dynamic range of abundances and molecular weights but is limited in its capacity to discriminate isobaric compounds. In this article, we have extended earlier studies using isotopic labeling for elemental composition elucidation (Rodgers, R. P.; Blumer, E. N.; Hendrickson, C. L.; Marshall, A. G. J. Am. Soc. Mass Spectrom. 2000, 11, 835-40) to limit the formulas consistent with any exact mass measurement by comparing observations of metabolites extracted from Arabidopsis thaliana plants grown with (I) (12)C and (14)N (natural abundance), (II) (12)C and (15)N, (III) (13)C and (14)N, or (IV) (13)C and (15)N. Unique elemental compositions were determined over a dramatically enhanced mass range by analyzing exact mass measurement data from the four extracts using two methods. In the first, metabolite masses were matched with a library of 11,000 compounds known to be present in living cells by using values calculated for each of the four isotopic conditions. In the second method, metabolite masses were searched against masses calculated for a constrained subset of possible atomic combinations in all four isotopic regimes. In both methods, the lists of elemental compositions from each labeling regime were compared to find common formulas with similar retention properties by HPLC in at least three of the four regimes. These results demonstrate that metabolic labeling can be used to provide additional constraints for higher confidence formula assignments over an extended mass range.
authors: Department of Biochemistry, University of Wisconsin, 433 Babcock Drive, Madison, Wisconsin 53706, USA.
authors: Adrian D
authors: AD
authors: Hegeman
authors: Christopher F
authors: CF
authors: Schulte
authors: Qiu
authors: Q
authors: Cui
authors: Ian A
authors: IA
authors: Lewis
authors: Edward L
authors: EL
authors: Huttlin
authors: Hamid
authors: H
authors: Eghbalnia
authors: Amy C
authors: AC
authors: Harms
authors: Eldon L
authors: EL
authors: Ulrich
authors: John L
authors: JL
authors: Markley
authors: Michael R
authors: MR
authors: Sussman
mesh_terms: D017360:Arabidopsis / Q000737:chemistry*; D002247:Carbon Isotopes; D002734:Chlorophyll / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D019992:Databases as Topic; D004602:Elements; D007553:Isotope Labeling; D013058:Mass Spectrometry; D000432:Methanol / Q000737:chemistry; D008962:Models, Theoretical; D015394:Molecular Structure; D009587:Nitrogen Isotopes; D018515:Plant Leaves / Q000737:chemistry* / Q000378:metabolism
chemical_list: D002247:Carbon Isotopes; D004602:Elements; D009587:Nitrogen Isotopes; D002734:Chlorophyll; D000432:Methanol