A method for measuring both glutamine and glutamate levels and stable isotopic enrichments

D. Darmaun, M. J. Manary, D. E. Matthews

Research output: Contribution to journalArticle

65 Scopus citations

Abstract

A method is described for measuring separately glutamine and glutamate levels and stable isotopic enrichment in plasma or whole blood samples by gas chromatography-mass spectrometry (GCMS). Deuterated internal standards are used for the quantitation via reverse isotope dilution and are added to plasma samples immediately upon sample collection. The samples are then applied to miniature anion-exchange columns to separate glutamine and glutamate, and the separated fractions are derivatized for GCMS analysis. The internal standards serve not only to quantitate both amino acids by reverse isotope dilution, but also to correct for glutamine deamidation to glutamate during sample storage and handling. Glutamine and glutamate are quantitated from plasma with typical precisions of 1 and 16%, respectively. Plasma glutamine and glutamate amino-15N enrichments are determined with precisions of 2 and 12%, respectively. The precision of the glutamate measurements for whole blood is typically 6%, where the glutamate levels are higher. This method uses inexpensive columns, allows simultaneous processing of multiple samples, and requires minimal volumes of plasma (250μl).

Original languageEnglish
Pages (from-to)92-102
Number of pages11
JournalAnalytical Biochemistry
Volume147
Issue number1
DOIs
StatePublished - May 15 1985

Keywords

  • amino acids
  • gas chromatography-mass spectrometry
  • isotope analysis
  • mass spectrometry
  • metabolites
  • nutrition

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