Quantitation of Positional isomers of deuterium-labeled glucose by gas chromatography/mass spectrometry

Guo, Zhao; Lee, W-N. Paul; Katz, Joseph; Bergner, A.

doi:10.1016/0003-2697(92)90238-3

Cited by 40 publications

(43 citation statements)

References 15 publications

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“…Since liver from fasted rats contain a small amount of glycogen, the amount actually found in the glycogen formed was closer to 1.0 (11). However, Guo et al (13), on giving 2H20 to fasted rats, found only 0.74 atom equivalents of deuterium bound to carbon 2 of blood glucose. In accord with the hydrogen bound to carbon 2 equilibrating with hydrogen in body water in our study, after enough time for the 2H20 to equilibrate, the enrichment of the hydrogen bound to carbon 2 was similar to the enrichment in water.…”

Section: Discussionmentioning

confidence: 99%

“…The use of 2H20 offers an alternative. However, there are also limits to the quantity of 2H20 that can be administered and a method adequate to quantify enrichment at carbon 6, as well as at carbon 2, at an acceptable dose of 2H20, is yet to be reported (12)(13)(14) present study reports such a method along with a presentation of its application to the fasted state. When glucose is oxidized by periodic acid, formaldehyde is formed bearing the hydrogens bound to carbon 6 of glucose.…”

Section: Introductionmentioning

confidence: 99%

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Use of 2H2O for estimating rates of gluconeogenesis. Application to the fasted state.

et al. 1995

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A method is introduced for estimating the contribution of gluconeogenesis to glucose production. 2H20 is administered orally to achieve 0.5% deuterium enrichment in body water. Enrichments are determined in the hydrogens bound to carbons 2 and 6 of blood glucose and in urinary water. Enrichment at carbon 6 of glucose is assayed in hexamethylenetetramine, formed from formaldehyde produced by periodate oxidation of the glucose. Enrichment at carbon 2 is assayed in lactate formed by enzymatic transfer of the hydrogen from glucose via sorbitol to pyruvate. The fraction gluconeogenesis contributes to glucose production equals the ratio of the enrichment at carbon 6 to that at carbon 2 or in urinary water. Applying the method, the contribution of gluconeogenesis in healthy subjects was 23-42% after fasting 14 h, increasing to 59-84% after fasting 42 h. Enrichment at carbon 2 to that in urinary water was 1.12±0.13. Therefore, the assumption that hydrogen equilibrated during hexose-6-P isomerization was fulfilled. The 3H/14C ratio in glucose formed from [3-3H,3-'4C]lactate given to healthy subjects was 0.1 to 0.2 of that in the lactate. Therefore equilibration during gluconeogenesis of the hydrogen bound to carbon 6 with that in body water was 80-90% complete, so that gluconeogenesis is underestimated by 10-20%. Glycerol's contribution to gluconeogenesis is not included in these estimates. The method is applicable to studies in humans of gluconeogenesis at safe doses of 21H20. (J. Clin. Invest. 1995. 95:172-178.)

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Use of 2H2O for estimating rates of gluconeogenesis. Application to the fasted state.

et al. 1995

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“…1A). After exposure to [U- 13 C 3 ]glycerol, the liver produces primarily [1,2,[3][4][5][6][7][8][9][10][11][12][13] C 3 ]glucose and [4,5,[6][7][8][9][10][11][12][13] C 3 ]glucose after equilibration of DHAP and GA3P through TPI. [U- 13 C 6 ]glucose is also produced, but it will not be considered further for simplicity because it is trivial in the current study.…”

Section: Asymmetry In Glucose Isotopomersmentioning

confidence: 99%

“…The triose phosphate isomerase (TPI) 2 reaction influences equilibration of tracer between carbons 1-3 and carbons 4 -6 of glucose. Essentially complete equilibration has been assumed in studies with hydrogen and carbon tracers (1)(2)(3) or confirmed experimentally in some preparations (4 -5). However, others find that equilibration is incomplete (6 -10).…”

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confidence: 91%

Interaction between the Pentose Phosphate Pathway and Gluconeogenesis from Glycerol in the Liver

Jin

Sherry

Malloy

2014

Journal of Biological Chemistry

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Background: Hepatic pentose phosphate pathway activity is difficult to monitor in vivo. Results: Hepatic pentose phosphate pathway results in different ratios of doubly labeled glucose isotopomers after administration of [U-13 C 3 ]glycerol. Conclusion: [1,2-13 C 2 ]glucose produced by the hepatic pentose phosphate pathway is quantified. Significance: A simple method is presented to detect the activity of hepatic pentose phosphate pathway in vivo from analysis of plasma glucose.

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“…This method quantifies rates of gluconeogenesis but requires patients to lie for hours within a magnet (7). An alternative method is based on incorporation of 2 H into glucose after 2 H 2 O administration (8). In humans, gluconeogenesis can be calculated from 2 H enrichments in hexamethylenetetramine (HMT) derived from blood glucose measured with gas chromatographymass spectrometry (9,10).…”

mentioning

confidence: 99%

Measurement of Fractional Whole-Body Gluconeogenesis in Humans From Blood Samples Using 2H Nuclear Magnetic Resonance Spectroscopy

Kunert

Stingl²,

Rosian³

et al. 2003

Diabetes

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Several problems limit quantification of gluconeogenesis. We applied in vitro 2 H-nuclear magnetic resonance (NMR) spectroscopy to simultaneously measure 2 H in all glucose carbons for direct assessment of gluconeogenesis. This method was compared with 2 H measurement in carbons 5 and 2 using gas chromatography-mass spectrometry (hexamethylenetetramine [HMT]) and with in vivo 13 C magnetic resonance spectroscopy (MRS). After 14 h of fasting, and following 2 H 2 O ingestion, blood was obtained from nine healthy and seven type 2 diabetic subjects. Glucose was purified, acetylated, and analyzed for 2 H in carbons 1-6 with 2 H-NMR. Using 5:2 ratios, gluconeogenesis increased (P < 0.05) over time and mean gluconeogenesis was lower in control subjects than in type 2 diabetic patients (63 ؎ 3 vs. 75 ؎ 2%, P < 0.01).13 C-MRS revealed higher hepatic glycogenolysis in control subjects (3.9 ؎ 0.4 vs. 2.3 ؎ 0.2 mol ⅐ kg ؊1 ⅐ min ؊1 ) yielding mean contribution of gluconeogenesis of 65 ؎ 3 and 77 ؎ 2% (P < 0.005). Measurement of gluconeogenesis by 2 H-NMR correlated linearly with 13 C-MRS (r ‫؍‬ 0.758, P ‫؍‬ 0.0007) and HMT (r ‫؍‬ 0.759, P ‫؍‬ 0.0007). In an additional protocol, 2 H enrichments demonstrated a fast decline of gluconeogenesis from ϳ100 to ϳ68% (P < 0.02) within 4 h of galactose infusion after 40 -44 h of fasting. Thus, in vitro 2 H-NMR offers an alternative approach to determine fractional gluconeogenesis in good agreement with standard methods and allows monitoring of rapid metabolic alterations.

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Quantitation of Positional isomers of deuterium-labeled glucose by gas chromatography/mass spectrometry

Cited by 40 publications

References 15 publications

Use of 2H2O for estimating rates of gluconeogenesis. Application to the fasted state.

Use of 2H2O for estimating rates of gluconeogenesis. Application to the fasted state.

Interaction between the Pentose Phosphate Pathway and Gluconeogenesis from Glycerol in the Liver

Measurement of Fractional Whole-Body Gluconeogenesis in Humans From Blood Samples Using 2H Nuclear Magnetic Resonance Spectroscopy

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