Two-dimensional liquid chromatography/mass spectrometry/mass spectrometry separation of water-soluble metabolites

Jacob N. Fairchild, Krisztian Horvath, Jessica R. Gooding, Shawn R. Campagna, Georges Guiochon

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)8161-8166
Number of pages6
JournalJournal of Chromatography A
Volume1217
Issue number52
DOIs
Publication statusPublished - Dec 24 2010

Keywords

  • Metabolites
  • Multidimensional chromatography
  • Off-line
  • Two-dimensional liquid chromatography

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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