Method for determining molar concentrations of metabolites in complex solutions from two-dimensional 1H-13C NMR spectra
Method for determining molar concentrations of metabolites in complex solutions from two-dimensional 1H-13C NMR spectra
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DOI：
10.1021/ac071583z
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发表时间：
2007-12-15
影响因子：
7.4
通讯作者：
Markley, John L.
中科院分区：
文献类型：
--
作者：
Lewis, Ian A.;Schommer, Seth C.;Markley, John L.
一维 (1D) H-1 核磁共振 (NMR) 波谱广泛用于生物体液和组织提取物中代谢物的高通量分析。通常，此类光谱被视为多元统计对象，而不是可量化代谢物的集合。我们在这里报告了一种二维 (2D) H-1-C-13 NMR 策略（快速代谢物定量，FMQ，通过 NMR），用于识别和定量生物样品中类似于 40 种最丰富的代谢物。为了验证这项技术，我们制备了合成化合物和拟南芥、酿酒酵母和苜蓿提取物的混合物。我们证明，使用我们的定量 2D H-1-C-13 NMR 策略可以在 12 分钟内确定准确的摩尔浓度（技术误差 2.7%）。相比之下，传统的 1D H-1 NMR 分析在接近理想的条件下会产生 16.2% 的技术误差。我们建议通过 NMR 进行 FMQ 作为 1D H-1 NMR 的实用替代方案，用于代谢组学研究，可以获得 50 mg（提取物干重）的样品。
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.