Method for determining molar concentrations of metabolites in complex solutions from two-dimensional 1H-13C NMR spectra

被引：217

作者：

Lewis, Ian A. ^{[1
]}

Schommer, Seth C. ^{[1
]}

Hodis, Brendan ^{[1
]}

Robb, Kate A. ^{[1
]}

Tonelli, Marco ^{[1
]}

Westler, William M. ^{[1
]}

Suissman, Michael R. ^{[1
]}

Markley, John L. ^{[1
]}

机构：

[1] Univ Wisconsin, Dept Biochem, Madison, WI 53706 USA

来源：

ANALYTICAL CHEMISTRY | 2007年 / 79卷 / 24期

关键词：

D O I：

10.1021/ac071583z

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

One-dimensional (1D) H-1 nuclear magnetic resonance (NMR) spectroscopy is used extensively for high-throughput analysis of metabolities in biological fluids and tissue extracts. Typically, such spectra are treated as multivariate statistical objects rather than as collections of quantifiable metabolites. We report here a two-dimensional (2D) H-1- C-13 NMR strategy (fast metabolite quantification, FMQ, by NMR) for identifying and quantifying the similar to 40 most abundant metabolites in biological samples. To validate this technique, we prepared mixtures of synthetic compounds and extracts from Arabidopsis thaliana, Saccharomyces cerevisiae, and Medicago sativa. We show that accurate (technical error 2.7%) molar concentrations can be determined in 12 min using our quantitative 2D H-1-C-13 NMR strategy. In contrast, traditional 1D H-1 NMR analysis resulted in 16.2% technical error under nearly ideal conditions. We propose FMQ by NMR as a practical alternative to 1D H-1 NMR for metabolomics studies in which 50-mg (extract dry weight) samples can be obtained.

引用

页码：9385 / 9390

页数：6

共 20 条

[11] Validation of quantitative NMR [J].