A carbonyl capture approach for profiling oxidized metabolites in cell extracts

Stephanie J. Mattingly, Tao Xu, Michael H. Nantz, Richard M. Higashi, Teresa W.M. Fan

Research output: Contribution to journalArticlepeer-review

27 Scopus citations

Abstract

Fourier-transform ion-cyclotron resonance mass spectrometry (FT-ICR-MS) detection of oxidized cellular metabolites is described using isotopologic, carbonyl-selective derivatizing agents that integrate aminooxy functionality for carbonyl capture, quaternary nitrogen for electrospray enhancement, and a hydrophobic domain for sample cleanup. These modular structural features enable rapid, sensitive analysis of complex mixtures of metabolite-derivatives by FT-ICR-MS via continuous nanoelectrospray infusion. Specifically, this approach can be used to globally assess levels of low abundance and labile aldehyde and ketone metabolites quantitatively and in high throughput manner. These metabolites are often key and unique indicators of various biochemical pathways and their perturbations. Analysis of lung adenocarcinoma A549 cells established a profile of carbonyl metabolites spanning multiple structural classes. We also demonstrate a procedure for metabolite quantification using pyruvate as a model analyte.

Original languageEnglish
Pages (from-to)989-996
Number of pages8
JournalMetabolomics
Volume8
Issue number6
DOIs
StatePublished - Dec 2012

Bibliographical note

Funding Information:
Acknowledgments We acknowledge financial support from NIH 1RO1CA118434-01A2 (TWMF), 3RO1CA118434-02S1 (TWMF), Kentucky Lung Cancer Research Program OGMB101380 (TWMF), and the University of Louisville Clinical & Translational Science Pilot Grant 20003. The FT-ICR-MS instrumentation at the Center for Regulatory and Environmental Analytical Metabolomics Mass Spectrometry Facility was funded by NSF/EPSCoR grant # EPS-0447479 (TWMF).

Keywords

  • Aldehyde
  • Cellular oxidation state
  • Ketone
  • Mass spectrometry
  • Metabolite profiling
  • Oxaloacetate
  • Pyruvate
  • α-ketoglutarate

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Clinical Biochemistry

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