Method for Determining Molar Concentrations of Metabolites in Complex Solutions from Two-Dimensional 1H-13C NMR Spectra

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• 作者：Ian A. Lewis ; Seth C. Schommer ; Brendan Hodis ; Kate A. Robb ; Marco Tonelli ; William M. Westler ; Michael R. Sussman ; John L. Markley
• 刊名：Analytical Chemistry
• 出版年：2007
• 出版时间：December 15, 2007
• 年：2007
• 卷：79
• 期：24
• 页码：9385 - 9390
• 全文大小：359K
• 年卷期：v.79,no.24(December 15, 2007)
• ISSN：1520-6882

文摘

One-dimensional (1D) ¹H nuclear magnetic resonance(NMR) spectroscopy is used extensively for high-throughput analysis of metabolites in biological fluids and tissueextracts. Typically, such spectra are treated as multivariatestatistical objects rather than as collections of quantifiablemetabolites. We report here a two-dimensional (2D) ¹H-¹³C NMR strategy (fast metabolite quantification, FMQ,by NMR) for identifying and quantifying the ~40 mostabundant metabolites in biological samples. To validatethis technique, we prepared mixtures of synthetic compounds and extracts from Arabidopsis thaliana, Saccharomyces cerevisiae, and Medicago sativa. We showthat accurate (technical error 2.7%) molar concentrationscan be determined in 12 min using our quantitative 2D¹H-¹³C NMR strategy. In contrast, traditional 1D ¹H NMRanalysis resulted in 16.2% technical error under nearlyideal conditions. We propose FMQ by NMR as a practicalalternative to 1D ¹H NMR for metabolomics studies inwhich 50-mg (extract dry weight) samples can be obtained.