Application of 13C isotope labeling using liquid chromatography mass spectrometry (LC-MS) to determining phosphate-containing metabolic incorporation

J Mass Spectrom. 2013 Dec;48(12):1270-5. doi: 10.1002/jms.3292.

Abstract

Here, we describe an approach wherein negative electrospray ionization mass spectrometry has used to understand the relative flux through phosphate containing metabolic intermediates associated with central carbon metabolism after administering cells with 13C-labeled substrates. The method was applied to examine the 13C incorporation through glycolysis in T47D breast cancer cells and showed reduction of glycolytic relative flux upon treatment with 2-Deoxyglucose.

Keywords: metabolic relative flux; single reaction monitoring; tandem mass spectrometry.

Publication types

  • Letter
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Breast Neoplasms / metabolism
  • Carbon Isotopes / analysis
  • Carbon Isotopes / metabolism*
  • Cell Line, Tumor
  • Chromatography, Liquid / methods
  • Female
  • Glycolysis / physiology
  • Humans
  • Isotope Labeling / methods*
  • Metabolic Flux Analysis / methods*
  • Phosphates / metabolism*
  • Spectrometry, Mass, Electrospray Ionization / methods
  • Tandem Mass Spectrometry

Substances

  • Carbon Isotopes
  • Phosphates