callahan-profilingof-2009.pdf (1.49 MB)
Profiling of polar metabolites in biological extracts using diamond hydride-based aqueous normal phase chromatography
journal contribution
posted on 2009-07-01, 00:00 authored by Damien CallahanDamien Callahan, D D Souza, A Bacic, U RoessnerHighly polar metabolites, such as sugars and most amino acids are not retained by conventional RP LC columns. Without sufficient retention low concentration compounds are not detected due ion suppression and structural isomers are not resolved. In contrast, hydrophilic interaction chromatography (HILIC) and aqueous normal phase chromatography (ANP) retain compounds based on their hydrophilicity and therefore provides a means of separating highly polar compounds. Here, an ANP method based on the diamond hydride stationary phase is presented for profiling biological small molecules by LC. A rapid separation system based upon a fast gradient that delivers reproducible chromatography is presented. Approximately 1000 compounds were reproducibly detected in human urine samples and clear differences between these samples were identified. This chromatography was also applied to xylem fluid from soyabean (Glycine max) plants to which 400 compounds were detected. This method greatly increases the metabolite coverage over RP-only metabolite profiling in biological samples. We show that both forms of chromatography are necessary for untargeted comprehensive metabolite profiling and that the diamond hydride stationary phase provides a good option for polar metabolite analysis.
History
Journal
Journal of separation scienceVolume
32Issue
13Pagination
2273 - 2280Publisher
WileyLocation
Londn, Eng.Publisher DOI
ISSN
1615-9306Language
engPublication classification
C Journal article; C1.1 Refereed article in a scholarly journalCopyright notice
2009, WileyUsage metrics
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No categories selectedKeywords
Aqueous normal phaseESIHydrophilic interaction chromatographyLC-MSMetabolomicsScience & TechnologyPhysical SciencesChemistry, AnalyticalChemistryHYDROPHILIC-INTERACTION CHROMATOGRAPHYINTERACTION LIQUID-CHROMATOGRAPHYIONIZATION MASS-SPECTROMETRYPRINCIPAL COMPONENT ANALYSISMETABONOMIC ANALYSISSTATIONARY-PHASERAT URINESEPARATIONMSCLASSIFICATION
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