Conditions for (13)C NMR detection of 2-hydroxyglutarate in tissue extracts from isocitrate dehydrogenase-mutated gliomas

Anal Biochem. 2015 Jul 15;481:4-6. doi: 10.1016/j.ab.2015.04.017. Epub 2015 Apr 20.


(13)C NMR (nuclear magnetic resonance) spectroscopy of extracts from patient tumor samples provides rich information about metabolism. However, in isocitrate dehydrogenase (IDH)-mutant gliomas, (13)C labeling is obscured in oncometabolite 2-hydroxyglutaric acid (2 HG) by glutamate and glutamine, prompting development of a simple method to resolve the metabolites. J-coupled multiplets in 2 HG were similar to glutamate and glutamine and could be clearly resolved at pH 6. A cryogenically cooled (13)C probe, but not J-resolved heteronuclear single quantum coherence spectroscopy, significantly improved detection of 2 HG. These methods enable the monitoring of (13)C-(13)C spin-spin couplings in 2 HG expressing IDH-mutant gliomas.

Keywords: (13)C NMR; 2-Hydroxyglutarate; Cryoprobe; IDH-mutant gliomas.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbon Isotopes / analysis
  • Glioma / genetics*
  • Glioma / pathology
  • Glutamic Acid / analysis
  • Glutamine / analysis
  • Glutarates / analysis*
  • Humans
  • Isocitrate Dehydrogenase / genetics*
  • Magnetic Resonance Spectroscopy / methods*
  • Mutation


  • Carbon Isotopes
  • Glutarates
  • Glutamine
  • alpha-hydroxyglutarate
  • Glutamic Acid
  • Isocitrate Dehydrogenase