Tracing gluconeogenesis with deuterated water: measurement of low deuterium enrichments on carbons 6 and 2 of glucose

Analytical Biochemistry
J W HazeyH Brunengraber

Abstract

The contribution of gluconeogenesis to glucose production in vivo can be measured by enriching body water with 0.5% 2H2O and measuring the glucose labeling ratio C6/C2 (Landau et al., J. Clin. Invest. 95, 172-178, 1995). We present further refinements of the measurements of the 2H enrichments on C6 and C2 of glucose. The transfer of 2H from C6 of glucose to hexamethylenetetramine (HMT) and extraction in preparation for gas chromatography-mass spectrometry can be done in a single test tube, without distillation of the intermediate formaldehyde. In addition, extraction of small amounts of HMT is greatly improved by making a HMT-iodine adduct. For C2, glucose is reduced to sorbitol, and 2H on C2 is transferred enzymatically to [U-13C3]pyruvate, forming [U-13C3,2-2H]lactate. The latter is assayed by negative chemical ionization gas chromatography-mass spectrometry of the pentafluorobenzyl derivative. The natural enrichment of the [U-13C3]lactyl ion is only 0.4%, allowing measurements of 2H enrichment down to 0.1%. These techniques were used in dogs infused with 2H2O and in isolated rat livers perfused with buffer containing 1 to 5% 2H2O. Our data reveal a difference in the rate of labeling of C6 and C2 of glucose in vivo. Lastly, i...Continue Reading

References

Jan 1, 1976·Current Topics in Cellular Regulation·J Katz, R Rognstad
Sep 1, 1974·European Journal of Biochemistry·R RognstadJ Katz
Jan 1, 1995·The Journal of Clinical Investigation·B R LandauS C Kalhan
Jul 15, 1996·The Journal of Clinical Investigation·B R LandauS C Kalhan
Jun 1, 1996·Journal of Mass Spectrometry : JMS·S F PrevisH Brunengraber

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Citations

Jan 1, 1997·Annual Review of Nutrition·H BrunengraberC Des Rosiers
Sep 5, 2002·Magnetic Resonance in Medicine : Official Journal of the Society of Magnetic Resonance in Medicine·John G JonesCarlos F G C Geraldes
Nov 24, 1999·Current Opinion in Clinical Nutrition and Metabolic Care·S F Previs, H Brunengraber
Mar 11, 2020·Nature Communications·Charandeep SinghJonathan E Sears

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