Dual Labeling of Metabolites for Metabolome Analysis (DLEMMA): A New Approach for the Identification and Relative Quantification of Metabolites by Means of Dual Isotope Labeling and Liquid Chromatography-Mass Spectrometry
Dual Labeling of Metabolites for Metabolome Analysis (DLEMMA): A New Approach for the Identification and Relative Quantification of Metabolites by Means of Dual Isotope Labeling and Liquid Chromatography-Mass Spectrometry
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DOI：
10.1021/ac901495a
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发表时间：
2009-11-15
影响因子：
7.4
通讯作者：
Aharoni, Asaph
中科院分区：
文献类型：
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作者：
Feldberg, Liron;Venger, Ilya;Aharoni, Asaph
预计先进的代谢组学技术将允许在全代谢组规模上鉴定和定量代谢物。然而，大多数代谢物要么仍然未知，要么无法明确识别。此外，由于样品制备和基质效应，目前的方法遭受相对定量的不准确性。在这里，我们提出了双标记代谢物的代谢组分析（DLEMMA）作为一个有价值的工具，它与DNA阵列分析类似，使鉴别和相对定量的差异代谢物在一个单一的样品。DLEMMA被证明是一种有效的方法，用于减少可能的化学结构分配，表现出相同的元素组成。它的力量体现在10种新型色氨酸衍生物的发现上。此外，通过饲喂两种苯丙氨酸标记的前体，我们可以利用DLEMMA检测转基因植物和对照植物之间的差异代谢产物。由于DLEMMA为两种样品提供了相同的基质，因此也提高了相对定量的准确性，从而避免了不同复杂生物基质对电喷雾电离的影响。因此，DLEMMA将补充和促进代谢组学技术的进步，并促进不同生物体的代谢途径发现。
Advanced metabolomics technologies are anticipated to permit the identification and quantification of metabolites at the whole-metabolome scale. Yet, most of the metabolites either remain unknown or cannot be identified unambiguously. Moreover, the present approaches suffer from inaccuracies in relative quantification because of sample preparation and matrix effects. Here we present Dual Labeling of Metabolites for Metabolome Analysis (DLEMMA) as a valuable tool, which with analogy to DNA array assays enables the identification and relative quantification of differential metabolites in a single sample. DLEMMA was demonstrated as an efficient method for reducing the number of possible chemical structures assigned that exhibit the same elemental composition. Its strength was exemplified by the discovery of 10 novel Tryptophan derivatives. Furthermore, employing DLEMMA by feeding two Phenylalanine-labeled precursors, we could detect differential metabolites between trans-genic and control plants. The accuracy of relative quantification is also enhanced since DLEMMA provides identical matrixes for both samples, thus avoiding the effects of different complex biological matrixes on electrospray ionization. Hence, DLEMMA will complement and contribute to the advancement of metabolomics technologies and boost metabolic pathway discovery in diverse organisms.