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Am J Physiol Endocrinol Metab (February 11, 2003). doi:10.1152/ajpendo.00485.2002
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Submitted on November 7, 2002
Accepted on February 3, 2003

MEASURING GLUCONEOGENESIS USING A LOW DOSE OF 2H2O: ADVANTAGE OF ISOTOPE FRACTIONATION DURING GAS CHROMATOGRAPHY

Jill Katanik1, Brendan J. McCabe1, Daniel Z. Brunengraber1, Visvanathan Chandramouli1, Fumie J Nishiyama2, Vernon E. Anderson3, and Stephen F. Previs1*

1 Department of Nutrition, Case Western Reserve University School of Medicine, Cleveland, OH, USA
2 Department of Medicine, Case Western Reserve University School of Medicine, Cleveland, OH, USA
3 Department of Biochemistry, Case Western Reserve University School of Medicine, Cleveland, OH, USA

* To whom correspondence should be addressed. E-mail: sxp29{at}po.cwru.edu.

The contribution of gluconeogenesis to glucose production can be measured by enriching body water with 2H2O to approximately 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 converting the corresponding glucose carbons to formaldehyde and then to hexamethylenetetraamine (HMT). We present a technique, for integrating ion chromatograms, which 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 and 2H-labeled HMT. We discuss the advantage(s) of using a low dose of 2H2O to quantify the contribution of gluconeogenesis.




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