Quantifying 13C-labeling in Free Sugars and Starch by GC-MS

  • Mohamed Koubaa
  • Brigitte Thomasset
  • Albrecht Roscher
Part of the Methods in Molecular Biology book series (MIMB, volume 1090)


We describe an approach to extract 13C-labeled sugars (glucose, fructose, maltose, sucrose, myo-inositol as well as glucose from starch) from plant tissues and to analyze their isotopomer distribution by gas chromatography–mass spectrometry (GC-MS). Sugars are derivatized with N,O-bis(trimethylsilyl) trifluoroacetamide (BSTFA) into their Si(CH3)3 derivatives. Electronic and chemical ionizations are used to obtain suitable fragments for metabolic flux analysis (MFA). Unique fragments are identified by computer simulation and experimental verification with labeled standards. Linear equations for separating information from glucosyl and fructosyl moieties of sucrose are presented. Finally, mass distributions are corrected for natural isotope abundance using a home-written program. The method is illustrated by sugar isotopomer analysis of 13C-labeled rapeseed embryos.

Key words

Metabolic flux analysis Isotopomers GC-MS 13C-labeling Sugars Brassica napus 


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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Mohamed Koubaa
    • 1
  • Brigitte Thomasset
    • 2
  • Albrecht Roscher
    • 3
  1. 1.Department of Molecular GeneticsThe Ohio State UniversityColumbusUSA
  2. 2.Universite de Technologie de Compiegne, CNRS-FRE 3580, Gènie Enzymatique et Cellulaire, Centre de Recherche de RoyallieuCompiègne CedexFrance
  3. 3.Génie Enzymatique et Cellulaire, FRE CNRS 3580Université de PicardieAmiens CedexFrance

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