The importance of accurately correcting for the natural abundance of stable isotopes

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• 作者：Firas S. Midani^a ; ^{firas.midani@duke.edu} ; Michelle L. Wynn^b ; ^c ; ^{mlwynn@umich.edu} ; Santiago Schnell^b ; ^{schnells@umich.edu}
• 关键词：Stable isotopes ; Natural abundance correction ; Metabolic flux analysis ; Reproducibility
• 刊名：Analytical Biochemistry
• 出版年：2017
• 出版时间：1 March 2017
• 年：2017
• 卷：520
• 期：Complete
• 页码：27-43
• 全文大小：1710 K
• 卷排序：520

文摘

The use of isotopically labeled tracer substrates is an experimental approach for measuring in vivo and in vitro intracellular metabolic dynamics. Stable isotopes that alter the mass but not the chemical behavior of a molecule are commonly used in isotope tracer studies. Because stable isotopes of some atoms naturally occur at non-negligible abundances, it is important to account for the natural abundance of these isotopes when analyzing data from isotope labeling experiments. Specifically, a distinction must be made between isotopes introduced experimentally via an isotopically labeled tracer and the isotopes naturally present at the start of an experiment. In this tutorial review, we explain the underlying theory of natural abundance correction of stable isotopes, a concept not always understood by metabolic researchers. We also provide a comparison of distinct methods for performing this correction and discuss natural abundance correction in the context of steady state 13C metabolic flux, a method increasingly used to infer intracellular metabolic flux from isotope experiments.