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The role of liquid chromatography and flow injection analyses coupled to isotope ratio mass spectrometry for studying human in vivo glucose metabolism

Authors


  • Presented at the Liquid Chromatography-Isotope Ratio Mass Spectrometry Users Meeting, held 23–24 November 2010 at the University of Oxford, Oxford, UK.

J.-P. Godin, Nestlé Research Center, Nestec, Ltd., PO Box 44, 1000 Lausanne 26, Switzerland.

E-mail: jean-philippe.godin@rdls.nestle.com

Abstract

Under most physiological conditions, glucose, or carbohydrate (CHO), homeostasis is tightly regulated. In order to mechanistically appraise the origin of circulating glucose (e.g. via either gluconeogenesis, glycogenolysis or oral glucose intake), and its regulation and oxidation, the use of stable isotope tracers is now a well-accepted analytical technique. Methodologically, liquid chromatography coupled to isotope ratio mass spectrometry (LC/IRMS) can replace gas chromatography coupled to combustion-isotope ratio mass spectrometry (GC/C/IRMS) for carrying out compound-specific 13C isotopic analysis. The LC/IRMS approach is well suited for studying glucose metabolism, since the plasma glucose concentration is relatively high and the glucose can readily undergo chromatography in an aqueous mobile phase. Herewith, we report two main methodological approaches in a single instrument: (1) the ability to measure the isotopic enrichment of plasma glucose to assess the efficacy of CHO-based treatment (cocoa-enriched) during cycling exercise with healthy subjects, and (2) the capacity to carry out bulk isotopic analysis of labeled solutions, which is generally performed with an elemental analyzer coupled to IRMS. For plasma samples measured by LC/IRMS the data show a isotopic precision SD(δ13C) and SD(APE) of 0.7 ‰ and 0.001, respectively, with δ13C and APE values of −25.48 ‰ and 0.06, respectively, being generated before and after tracer administration. For bulk isotopic measurements, the data show that the presence of organic compounds in the blank slightly affects the δ13C values. Despite some analytical limitations, we clearly demonstrate the usefulness of the LC/IRMS especially when 13C-glucose is required during whole-body human nutritional studies. Copyright © 2011 John Wiley & Sons, Ltd.

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