Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1217, Issue 52, Pages 8161-8166Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2010.10.068
Keywords
Two-dimensional liquid chromatography; Multidimensional chromatography; Off-line; Metabolites
Funding
- US Department of Energy [DE-FG05-88-ER-13869]
- University of Tennessee
- Oak Ridge National Laboratory
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Off-line two-dimensional liquid chromatography with tandem mass spectrometry detection (2D-LC/MS-MS) was used to separate a set of metabolomic species. Water-soluble metabolites were extracted from Escherichia coli and Saccharomyces cerevisae cultures and were immediately analyzed using strong cation exchange (SCX)-hydrophilic interaction chromatography (HILIC). Metabolite mixtures are well-suited for multidimensional chromatography as the range of components varies widely with respect to polarity and chemical makeup. Some currently used methods employ two different separations for the detection of positively and negatively ionized metabolites by mass spectrometry. Here we developed a single set of chromatographic conditions for both ionization modes and were able to detect a total of 141 extracted metabolite species, with an overall peak capacity of ca. 2500. We show that a single two-dimensional separation method is sufficient and practical when a pair or more of unidimensional separations are used in metabolomics. (C) 2010 Elsevier B.V. All rights reserved.
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