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Am J Physiol Endocrinol Metab 284: E1043-E1048, 2003. First published February 11, 2003; doi:10.1152/ajpendo.00485.2002
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Vol. 284, Issue 5, E1043-E1048, May 2003

SPECIAL COMMUNICATIONS
Measuring gluconeogenesis using a low dose of 2H2O: advantage of isotope fractionation during gas chromatography

Jill Katanik1, Brendan J. McCabe1, Daniel Z. Brunengraber1, Visvanathan Chandramouli3, Fumie J. Nishiyama3, Vernon E. Anderson2, and Stephen F. Previs1

Departments of 2 Biochemistry, 3 Medicine, and 1 Nutrition, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106

The contribution of gluconeogenesis to glucose production can be measured by enriching body water with 2H2O to ~0.5% 2H and determining the ratio of 2H that is bound to carbon-5 vs. carbon-2 of blood glucose. This labeling ratio can be measured using gas chromatography-mass spectrometry after the corresponding glucose carbons are converted to formaldehyde and then to hexamethylenetetramine (HMT). We present a technique for integrating ion chromatograms that allows one to use only 0.05% 2H in body water (i.e., 10 times less than the current dose). This technique takes advantage of the difference in gas chromatographic retention times of naturally labeled HMT and [2H]HMT. We discuss the advantage(s) of using a low dose of 2H2O to quantify the contribution of gluconeogenesis.

diabetes; stable isotopes; gas chromatography-mass spectrometry


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