Clinical Chemistry
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Clinical Chemistry 55: 527-532, 2009. First published January 30, 2009; 10.1373/clinchem.2008.113654
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
clinchem.2008.113654v1
55/3/527    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Zhang, H.
Right arrow Articles by Millington, D. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zhang, H.
Right arrow Articles by Millington, D. S.
(Clinical Chemistry. 2009;55:527-532.)
© 2009 American Association for Clinical Chemistry, Inc.

Endocrinology and Metabolism

A Convenient LC-MS Method for Assessment of Glucose Kinetics In Vivo with D-[13C6]Glucose as a Tracer

Haoyue Zhang1, Robert D. Stevens2, Sarah P. Young1, Richard Surwit3, Anastasia Georgiades3, Raymond Boston4 and David S. Millington1,a

1 Department of Pediatrics, Medical Genetics Division, 2 Stedman Center for Nutrition, and 3 Department of Psychiatry and Behavioral Sciences, Medical Psychology Division, Duke University Medical Center, Durham, NC; 4 School of Veterinary Medicine, University of Pennsylvania, Kennet Square, PA.

aAddress correspondence to this author at: DUMC Biochemical Genetics Laboratory, 801 Capitola Dr. Suite 6, Durham, NC 27713. Fax 919-549-0709; e-mail dmilli{at}duke.edu.

Background: The isotope-labeled intravenous glucose tolerance test (IVGTT) combined with computer modeling is widely used to derive parameters related to glucose metabolism in vivo. Most of these methods involve use of either 2H2-labeled or 13C1-labeled D-glucose as a tracer with GC-MS to measure the isotope enrichment. These methods are challenging, both technologically and economically. We have developed a novel approach that is suitable for labeled-IVGTT studies involving a large cohort of individuals.

Methods: The tracer, D-[13C6]glucose, is a low-cost alternative with the significant advantage that the sixth isotope of natural glucose has virtually zero natural abundance, which facilitates isotopomer analysis with <1% labeled glucose in the infusate. After deproteinization of plasma samples collected at various times, glucose is converted to a stable derivative, purified by solid-phase extraction (SPE), and analyzed by HPLC–electrospray ionization mass spectrometry to accumulate the isotope-abundance data for the A+2, A+3, and A+6 ions of the glucose derivative. A 2-pool modeling program was used to derive standard kinetic parameters.

Results: With labeled-IVGTT data from 10 healthy male individuals, the values for insulin sensitivity, glucose effectiveness, and the plasma clearance rate estimated with the 2-pool minimal model compared well with values obtained via traditional methods.

Conclusions: The relative simplicity and robustness of the new method permit the preparation and analysis of up to 48 samples/day, a throughput equivalent to 2 complete IVGTT experiments, and this method is readily adaptable to existing 96 well–format purification and analytical systems.






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2009 by the American Association for Clinical Chemistry.