title: A novel ion pairing LC/MS metabolomics protocol for study of a variety of biologically relevant polar metabolites.
abstract: We report a method of ion-pairing liquid chromatography coupled to mass spectrometry (IP-LC-MS) that we have developed for the sensitive detection and quantification of a variety of biologically relevant polar molecules. We use the ion-pairing agent diamyl ammonium to improve chromatographic resolution of polar compounds, such as nucleotide cofactors, sugar phosphates, and organic acids, that are generally poorly retained by conventional reverse phase chromatographic methods. This method showed good linearity (average R value of 0.996) and reproducibility (generally RSD values <10%). We demonstrate the utility of this method by investigating the metabolomic signature of three distinct biological systems: the metabolic response to lack of superoxide dismutase activity and to paraquat induced oxidative stress, and the metabolic profiles of four different Drosophila species.
authors.affiliation: Department of Chemistry & Biochemistry, Laurentian University, Canada.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Knee
authors.forename: Teresa Z
authors.initials: TZ
authors.lastname: Rzezniczak
authors.forename: Aiko
authors.initials: A
authors.lastname: Barsch
authors.forename: Kevin Z
authors.initials: KZ
authors.lastname: Guo
authors.forename: Thomas J S
authors.initials: TJ
authors.lastname: Merritt
mesh_terms: D000085:Acetates; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D004330:Drosophila / Q000737:chemistry; D005260:Female; D057927:Hydrophobic and Hydrophilic Interactions; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D055442:Metabolome; D055432:Metabolomics / Q000379:methods*; D008956:Models, Chemical; D015203:Reproducibility of Results
chemical_list: D000085:Acetates; C018824:ammonium acetate
keywords: Diamyl ammonium (DAA); Drosophila metabolomics; IP-LC–MS; Ion pairing chromatography; Polar metabolites