Design of experiments:: an efficient strategy to identify factors influencing extraction and derivatization of Arabidopsis thaliana samples in metabolomic studies with gas chromatography/mass spectrometry
Design of experiments:: an efficient strategy to identify factors influencing extraction and derivatization of Arabidopsis thaliana samples in metabolomic studies with gas chromatography/mass spectrometry
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DOI：
10.1016/j.ab.2004.04.037
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发表时间：
2004-08-15
影响因子：
2.9
通讯作者：
Moritz, T
中科院分区：
文献类型：
--
作者：
Gullberg, J;Jonsson, P;Moritz, T
代谢组学研究的通常目的是量化一系列生物样品中每个样品的整个代谢组。为了对复杂的生物基质做到这一点，例如，植物组织，必须开发有效的和可重复的提取方案。然而，如果涉及GC/MS（代谢组学最广泛使用的分析方法之一），还必须制定衍生化方案。本研究的目的是探讨弓不同的化学和物理因素（提取溶剂，衍生试剂，温度）的影响提取和衍生化的植物拟南芥叶片的代谢物组。采用实验设计方法，系统地引入了变异，并利用回归模型分析了变异的影响。结果表明，这种方法允许一个可靠的协议，拟南芥代谢组学分析，以确定与相对有限数量的实验。在两次不同的研究之后，选择了提取和衍生化方案。此外，对66种内源性化合物分析的再现性进行了研究，结果表明，亲水性和亲脂性化合物均以高再现性检测到。(C)2004年爱思唯尔公司All rights reserved.
The usual aim in metabolomic studies is to quantify the entire metabolome of each of a series of biological samples. To do this for complex biological matrices, e.g., plant tissues, efficient and reproducible extraction protocols must be developed. However, derivatization protocols must also be developed if GC/MS (one of the mostly widely used analytical methods for metabolomics) is involved. The aim of this study was to investigate bow different chemical and physical factors (extraction solvent, derivatization reagents, and temperature) affect the extraction and derivatization of the metabolome from leaves of the plant Arabidopsis thaliana. Using design of experiment procedures, variation was systematically introduced, and the effects of this variation were analyzed using regression models. The results show that this approach allows a reliable protocol for metabolomic analysis of Arabidopsis to be determined with a relatively limited number of experiments. Following two different investigations an extraction and derivatization protocol was chosen. Further, the reproducibility of the analysis of 66 endogenous compounds was investigated, and it was shown that both hydrophilic and lipophilic compounds were detected with high reproducibility. (C) 2004 Elsevier Inc. All rights reserved.