title: Detection of 2-hydroxyglutarate in formalin-fixed paraffin-embedded glioma specimens by gas chromatography/mass spectrometry.
abstract: Mutations in the isocitrate dehydrogenase (IDH) 1 and 2 genes occur frequently in diffuse astrocytoma and oligodendroglioma. The consecutive amino acid substitutions in the mutant proteins result in a gain of the function to catalyze the reduction of alpha-ketoglutarate to 2-hydroxyglutarate (2HG). So far, all investigated IDH mutations share this gain of function. We here describe a method to detect 2HG levels in archival formalin-fixed paraffin-embedded tumor specimens by stable isotope dilution using gas chromatography followed by mass spectrometry (GC/MS). While 2HG levels are notably decreased during the routine embedding process, preserved amounts are still sufficient to indicate a mutation. Detection of 2HG in archival specimens could make routinely processed tissue accessible for research on 2HG accumulation and may allow studies on correlation with clinical data.
authors.affiliation: Department of Neuropathology, Institute of Pathology, Ruprecht-Karls-University Heidelberg, Heidelberg.
authors.forename: Felix
authors.initials: F
authors.lastname: Sahm
authors.forename: David
authors.initials: D
authors.lastname: Capper
authors.forename: Stefan
authors.initials: S
authors.lastname: Pusch
authors.forename: Jörg
authors.initials: J
authors.lastname: Balss
authors.forename: Arend
authors.initials: A
authors.lastname: Koch
authors.forename: Claus-Dieter
authors.initials: CD
authors.lastname: Langhans
authors.forename: Jürgen G
authors.initials: JG
authors.lastname: Okun
authors.forename: Andreas
authors.initials: A
authors.lastname: von Deimling
mesh_terms: D001932:Brain Neoplasms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005404:Fixatives; D005557:Formaldehyde; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D005910:Glioma / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005977:Glutarates / Q000378:metabolism*; D006801:Humans; D010232:Paraffin; D016612:Paraffin Embedding / Q000379:methods; D016707:Tissue Fixation / Q000379:methods
chemical_list: D005404:Fixatives; D005977:Glutarates; D005557:Formaldehyde; C019417:alpha-hydroxyglutarate; D010232:Paraffin