Open AccessA New Method for the Determination of Specific 13 C Enrichment in Phosphorylated [1– 13 C]glucose Metabolites
Author(s) -
Lutz Norbert W.,
Yahi Nouara,
Fantini Jacques,
Cozzone Patrick J.
Publication year - 1996
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1996.0470z.x
Subject(s) - perchloric acid , chemistry , nuclear magnetic resonance spectroscopy , biochemistry , carbon 13 nmr , metabolite , metabolic pathway , glycolysis , anabolism , metabolism , chromatography , stereochemistry , organic chemistry
A 31 P‐NMR method for the determination of 13 C enrichment in phosphorylated [1‐ 13 C]glucose metabolites was developed by taking advantage of the 13 C satellites detectable for 31 P‐NMR signals of metabolites such as UDP‐hexoses, UDP‐ N ‐acetylhexosamines and other phosphorylated compounds generated during glycolysis and subsequent anabolism. HT‐29 cells were incubated in culture medium containing 4.5 g/l [1‐ 13 C]glucose for 24 h prior to cell extraction, and high‐resolution 31 P‐NMR spectra were acquired from perchloric acid extracts. Since glucose and its phosphorylated products are key metabolites for many different metabolic processes, this method may be very helpful for studying specific metabolic pathways involving phosphorylated glucose metabolites.