[13C] GC-C-IRMS analysis of methylboronic acid derivatives of glucose from liver glycogen after the ingestion of [13C] labeled tracers in rats

J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Nov 1;877(29):3638-44. doi: 10.1016/j.jchromb.2009.09.005. Epub 2009 Sep 10.


We developed a complete method to measure low [(13)C] enrichments in glycogen. Fourteen rats were fed a control diet. Six of them also ingested either [U-(13)C] glucose (n=2) or a mixture of 20 [U-(13)C] amino acids (n=4). Hepatic glycogen was extracted, digested to glucose and purified on anion-cation exchange resins. After the optimization of methylboronic acid derivatization using GC-MS, [(13)C] enrichment of extracted glucose was measured by GC-C-IRMS. The accuracy was addressed by measuring the enrichment excess of a calibration curve, which observed values were in good agreement with the expected values (R=0.9979). Corrected delta values were -15.6+/-1.6 delta(13)C (per thousand) for control rats (n=8) and increased to -5 to 8 delta(13)C (per thousand) per thousand and 12-14 delta(13)C (per thousand) per thousand after the ingestion of [U-(13)C] amino acids or [U-(13)C] glucose as oral tracers, respectively. The method enabled the determination of dietary substrate transfer into glycogen. The sequestration of dietary glucose in liver glycogen 4 h after the meal was 35% of the ingested dose whereas the transfer of carbon skeletons from amino acids was only 0.25 to 1%.

MeSH terms

  • Animals
  • Boron Compounds / chemistry*
  • Carbon Isotopes / analysis*
  • Gas Chromatography-Mass Spectrometry / methods*
  • Glucose / chemistry*
  • Liver Glycogen / chemistry
  • Liver Glycogen / metabolism*
  • Male
  • Molecular Structure
  • Rats
  • Rats, Wistar


  • Boron Compounds
  • Carbon Isotopes
  • Liver Glycogen
  • methaneboronic acid
  • Glucose