A Method for Obtaining 13C Isotopomer Populations in 13C-Enriched Glucose

J. G. Jones, G. L. Cottam, B. C. Miller, A. D. Sherry, C. R. Malloy

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Abstract

13C NMR analysis of 13C-enriched glucose containing multiple isotopomers is hampered by chemical shift similarities of several carbon resonances and by the presence of two anomeric forms. A convenient and quantitative method of enzymatically oxidizing glucose to gluconate in tissue and perfusate extracts is presented. The six carbon resonances of the resulting 13C-enriched gluconate are fully resolved at high pH, thereby allowing a determination of the fractional population of each 13C isotopomer by 13C NMR. The utility of this method is demonstrated using the effluent from an isolated perfused liver containing 13C-enriched glucose produced by hepatic metabolism of sodium [1,2,3-13C3]propionate via the citric acid cycle and gluconeogenesis. An analysis of the gluconate C2 and C5 resonances in this sample showed that pentose phosphate activity was insignificant during this perfusion protocol. As demonstrated, this method provides a means of fully describing 13C isotopomer populations in enriched glucose samples where isotope may be derived from multiple metabolic pathways, thus expanding the scope of experimental design and enrichment strategies.

Original languageEnglish (US)
Pages (from-to)148-152
Number of pages5
JournalAnalytical Biochemistry
Volume217
Issue number1
DOIs
StatePublished - Feb 15 1994

Fingerprint

Glucose
Population
Carbon
Nuclear magnetic resonance
Pentoses
Tissue Extracts
Citric Acid Cycle
Gluconeogenesis
Liver
Propionates
Chemical shift
Metabolic Networks and Pathways
Metabolism
Isotopes
Design of experiments
Effluents
Research Design
Perfusion
Sodium
Phosphates

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Biochemistry

Cite this

A Method for Obtaining 13C Isotopomer Populations in 13C-Enriched Glucose. / Jones, J. G.; Cottam, G. L.; Miller, B. C.; Sherry, A. D.; Malloy, C. R.

In: Analytical Biochemistry, Vol. 217, No. 1, 15.02.1994, p. 148-152.

Research output: Contribution to journalArticle

Jones, J. G. ; Cottam, G. L. ; Miller, B. C. ; Sherry, A. D. ; Malloy, C. R. / A Method for Obtaining 13C Isotopomer Populations in 13C-Enriched Glucose. In: Analytical Biochemistry. 1994 ; Vol. 217, No. 1. pp. 148-152.
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