✦ LIBER ✦

Quantitation of Gluconeogenesis by 2H Nuclear Magnetic Resonance Analysis of Plasma Glucose Following Ingestion of 2H2O

✍ Scribed by John G. Jones; Rui A. Carvalho; A.Dean Sherry; Craig R. Malloy

Publisher: Elsevier Science
Year: 2000
Tongue: English
Weight: 79 KB
Volume: 277
Category: Article
ISSN: 0003-2697
DOI: 10.1006/abio.1999.4391

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✦ Synopsis

We present a simple 2 H NMR assay of the fractional contribution of gluconeogenesis to hepatic glucose output following ingestion of 2 H 2 O. The assay is based on the measurement of relative deuterium enrichment in hydrogens 2 and 3 of plasma glucose. Plasma glucose was enzymatically converted to gluconate, which displays fully resolved deuterium 2 and 3 resonances in its 2 H NMR spectrum at 14.1 T. The signal intensity of deuterium 3 relative to deuterium 2 in the gluconate derivative as quantitated by 2 H NMR was shown to provide a precise and accurate measurement of glucose enrichment in hydrogen 3 relative to hydrogen 2. This measurement was used to estimate the fractional contribution of gluconeogenesis to hepatic glucose output for two groups of rats; one group was fasted for 7 h and the other was fasted for 29 h. Rats were administered 2 H 2 O to enrich total body water to 5% over the last 4 -5 h of each fasting period. For the 7-h fasted group, the hydrogen 3/hydrogen 2 enrichment ratio of plasma glucose was 0.32 ؎ 0.09 (n ‫؍‬ 7). This indicates that gluconeogenesis contributed 32 ؎ 9% of total hepatic glucose output with glycogenolysis contributing the remainder. For the 29-h fasted group, the hydrogen 3/hydrogen 2 enrichment ratio of plasma glucose was 0.81 ؎ 0.10 (n ‫؍‬ 6), indicating that gluconeogenesis supplied the bulk of hepatic glucose output (81 ؎ 10%).

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