Use of [6,6- 2H 2]Glucose and of Low-Enrichment [U- 13C 6]-Glucose for Sequential or Simultaneous Measurements of Glucose Turnover by Gas Chromatography-Mass Spectrometry

We developed gas chromatography-mass spectrometric methods for assaying the enrichment of 99 at.% [6,6- 2H 2]glucose and 30 at.% [U- 13C 6]glucose, although both tracers are mostly M + 2. 13C enrichment is determined either by the C-1 to C-5 fragment of glucose aldonitrile pentaacetate or by oxidati...

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Bibliographic Details
Published in:Analytical biochemistry 1994-04, Vol.218 (1), p.192-196
Main Authors: Previs, S.F., Ciraolo, S.T., Fernandez, C.A., Beylot, M., Agarwal, K.C., Soloviev, M.V., Brunengraber, H.
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Animals
Biological and medical sciences
Carbohydrates
Carbon Isotopes
Deuterium
Fundamental and applied biological sciences. Psychology
Gas Chromatography-Mass Spectrometry
Glucaric Acid - chemistry
Glucose - analogs & derivatives
Glucose - analysis
Glucose - metabolism
Nitriles - analysis
Oses
Other biological molecules
Oxidation-Reduction
Radioligand Assay
Rats
Rats, Sprague-Dawley
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Summary:We developed gas chromatography-mass spectrometric methods for assaying the enrichment of 99 at.% [6,6- 2H 2]glucose and 30 at.% [U- 13C 6]glucose, although both tracers are mostly M + 2. 13C enrichment is determined either by the C-1 to C-5 fragment of glucose aldonitrile pentaacetate or by oxidation of glucose to glucarate. 2H enrichment is assayed as the difference between the 13C enrichment of glucarate and the 2H + 13C enrichment of glucose. The techniques, which were validated in in vivo experiments, are applicable to the determination of simultaneous or sequential measurements of the rate of glucose appearance before and after an intervention. They could also be applied to the simultaneous determination of (i) gluconeogenesis by incorporation of a 13C-labeled precursor into glucose and (ii) the rate of glucose appearance by [6,6- 2H 2]glucose infusion.
ISSN:0003-2697
1096-0309
DOI:10.1006/abio.1994.1159