Measurement of 2-hydroxyglutarate enantiomers in serum by chiral gas chromatography-tandem mass spectrometry and its application as a biomarker for IDH mutant gliomas

Shinji K. Strain, Morris D. Groves, Kelly L. Olino, Mark R. Emmett

Research output: Contribution to journalArticle

Abstract

(R)-2-hydroxyglutarate (R-2-hg) is a metabolite produced under physiologic conditions but also by tumors harboring isocitrate dehydrogenase (IDH) mutations. Detection is challenging as it must be distinguished from its enantiomer (S)-2-hydroxyglutarate (S-2-hg), which is also produced in the body but can increase under hypoxic conditions. A chiral gas chromatography-tandem mass spectrometry (GC–MS/MS) assay was developed, which separated enantiomers using a chiral column and quantified levels using a stable-isotope internal standard. The assay improves upon current methods by avoiding chiral derivatization and implementing a simplified sample extraction procedure. The assay was validated and serum 2-hg levels from healthy patients were measured, establishing a new, comprehensive reference range for normal levels of each enantiomer. Differences in basal levels were observed between races, but not sex. Age also correlated with S-2-hg levels, but not R-2-hg levels. Finally, serum levels of 2-hg enantiomers were measured in a pilot study of 11 patients with and without IDH mutant gliomas. An increase in R-2-hg levels was observed in 2/3 patients with actively growing IDH mutant gliomas. S-2-hg levels were increased in 4/11 patients, irrespective of IDH status. The results presented demonstrate the feasibility of a GC–MS/MS assay for measurement of 2-hg enantiomer levels for clinical use.

Original languageEnglish (US)
Pages (from-to)16-24
Number of pages9
JournalClinical Mass Spectrometry
Volume15
DOIs
StatePublished - Jan 2020

Fingerprint

Isocitrate Dehydrogenase
Enantiomers
Biomarkers
Gas chromatography
Mass spectrometry
Assays
Metabolites
Isotopes
alpha-hydroxyglutarate
Tumors

Keywords

  • 2-Hydroxyglutarate enantiomers
  • Gas-Chromatography-Mass Spectrometry, Enantioseparation
  • Isocitrate dehydrogenase mutation
  • Serum

ASJC Scopus subject areas

  • Spectroscopy

Cite this

Measurement of 2-hydroxyglutarate enantiomers in serum by chiral gas chromatography-tandem mass spectrometry and its application as a biomarker for IDH mutant gliomas. / Strain, Shinji K.; Groves, Morris D.; Olino, Kelly L.; Emmett, Mark R.

In: Clinical Mass Spectrometry, Vol. 15, 01.2020, p. 16-24.

Research output: Contribution to journalArticle

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AB - (R)-2-hydroxyglutarate (R-2-hg) is a metabolite produced under physiologic conditions but also by tumors harboring isocitrate dehydrogenase (IDH) mutations. Detection is challenging as it must be distinguished from its enantiomer (S)-2-hydroxyglutarate (S-2-hg), which is also produced in the body but can increase under hypoxic conditions. A chiral gas chromatography-tandem mass spectrometry (GC–MS/MS) assay was developed, which separated enantiomers using a chiral column and quantified levels using a stable-isotope internal standard. The assay improves upon current methods by avoiding chiral derivatization and implementing a simplified sample extraction procedure. The assay was validated and serum 2-hg levels from healthy patients were measured, establishing a new, comprehensive reference range for normal levels of each enantiomer. Differences in basal levels were observed between races, but not sex. Age also correlated with S-2-hg levels, but not R-2-hg levels. Finally, serum levels of 2-hg enantiomers were measured in a pilot study of 11 patients with and without IDH mutant gliomas. An increase in R-2-hg levels was observed in 2/3 patients with actively growing IDH mutant gliomas. S-2-hg levels were increased in 4/11 patients, irrespective of IDH status. The results presented demonstrate the feasibility of a GC–MS/MS assay for measurement of 2-hg enantiomer levels for clinical use.

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