Metabolomic analysis of mammalian cells and human tissue through one-pot two stage derivatizations using sheathless capillary electrophoresis-electrospray ionization-mass spectrometry

Tianjiao Huang, Michael Armbruster, Richard Lee, Dawn S. Hui, James L. Edwards

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Analysis of metabolites is often performed using separations coupled to mass spectrometry which is challenging due to their vast structural heterogeneity and variable charge states. Metabolites are often separated based on their class/functional group which in large part determine their acidity or basicity. This charge state dictates the ionization mode and efficiency of the molecule. To improve the sensitivity and expand the coverage of the mammalian metabolome, multifunctional derivatization with sheathless CE-ESI-MS was undertaken. In this work, amines, hydroxyls and carboxylates were labeled with tertiary amines tags. This derivatization was performed in under 100 min and resulted in high positive charge states for all analytes investigated. Amino acids and organic acids showed average limits of detection of 76 nM with good linearity of 0.96 and 10% RSD for peak area. Applying this metabolomic profiling system to bovine aortic endothelial cells showed changes in 15 metabolites after treatment with high glucose. The sample injection volume on-capillary was <300 cells for quantitative analyses. Targeted metabolites were found in human tissue, which indicates possible application of the system complex metabolome quantitation.

Original languageEnglish (US)
Pages (from-to)219-225
Number of pages7
JournalJournal of Chromatography A
Volume1567
DOIs
StatePublished - Sep 14 2018

Fingerprint

Capillary electrophoresis
Electrospray ionization
Metabolomics
Metabolome
Electrospray Ionization Mass Spectrometry
Capillary Electrophoresis
Metabolites
Amines
Mass spectrometry
Cells
Tissue
Hydroxyl Radical
Limit of Detection
Mass Spectrometry
Endothelial Cells
Amino Acids
Glucose
Injections
Acids
Organic acids

Keywords

  • Derivatization
  • Diabetes
  • Hyperglycemia
  • Metabolomics
  • Tissue

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Cite this

Metabolomic analysis of mammalian cells and human tissue through one-pot two stage derivatizations using sheathless capillary electrophoresis-electrospray ionization-mass spectrometry. / Huang, Tianjiao; Armbruster, Michael; Lee, Richard; Hui, Dawn S.; Edwards, James L.

In: Journal of Chromatography A, Vol. 1567, 14.09.2018, p. 219-225.

Research output: Contribution to journalArticle

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