title: Analysis of glycolytic intermediates with ion chromatography- and gas chromatography-mass spectrometry.
abstract: In this chapter, we describe a method for the quantitative analysis of glycolytic intermediates using ion chromatography-mass spectrometry (IC-MS) and gas chromatography (GC)-MS as complementary methods. With IC-MS-MS, pyruvate, glucose-6-phosphate, fructuse-6-phosphate, fructose-1,6-bisphosphate, phosphoenolpyruvate, and the sum of 2-phosphoglyceraldehyde + 3-phosphoglyceraldehyde can be quantified. With GC-MS using selected ion monitoring, glyceraldehyde-3-phosphate, dihydroxyacetonephosphate, 2-phosphoglyceraldehyde, and 3-phosphoglyceraldehyde can be analyzed. The derivatization for GC-MS is performed in two steps. In the first step, the keto and the aldehyde groups are oximated. In the next step, a silylation with N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) is performed, giving TMS-MOX derivatives of the glycolytic intermediates. The derivatives are separated on a GC column and detected with MS in SIM mode.
authors.affiliation: Department of Biotechnology, Delft University of Technology, Delft, The Netherlands. j.c.vanDam@tudelft.nl
authors.forename: Jan C
authors.initials: JC
authors.lastname: van Dam
authors.forename: Cor
authors.initials: C
authors.lastname: Ras
authors.forename: Angela
authors.initials: A
authors.lastname: ten Pierick
mesh_terms: D002241:Carbohydrates / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D002852:Chromatography, Ion Exchange / Q000379:methods* / Q000592:standards; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods* / Q000592:standards; D006019:Glycolysis; D012015:Reference Standards
chemical_list: D002241:Carbohydrates