Extraction and GC/MS analysis of the human blood plasma metabolome
Extraction and GC/MS analysis of the human blood plasma metabolome
复制标题
DOI：
10.1021/ac051211v
复制
发表时间：
2005-12-15
影响因子：
7.4
通讯作者：
Moritz, T
中科院分区：
文献类型：
--
作者：
Jiye, A;Trygg, J;Moritz, T
分析整个低分子量化合物（LMC），代谢组，可以为疾病的机制和新的诊断标志物提供更深入的见解。在调查中，我们采用实验设计理论（实验设计），制定了一种提取衍生化方案，用于GC/MS分析人血浆代谢组。通过使用多元统计工具（包括主成分分析和潜在结构的偏最小二乘投影）评估500多个已分辨峰的数据，对方案进行了优化。考察了甲醇、乙醇、乙腈、丙酮、氯仿五种有机溶剂单独和联合提取的性能，以优化提取效果。PLS分析表明甲醇萃取效率高，重现性好。优化了提取条件和衍生化条件。32种内源化合物的定量数据具有良好的精度和线性。此外，8种选定化合物的测定量与经认可实验室独立方法的分析结果吻合良好，并且大多数化合物可以在类似于0.1 pmol注射的绝对水平下检测到，对应于0.1至1 μ m的血浆浓度。结果表明，该方法可以有效地整合到代谢组学研究中，用于各种目的，例如识别与疾病相关的生物标志物。
Analysis of the entire set of low molecular weight compounds (LMC), the metabolome, could provide deeper insights into mechanisms of disease and novel markers for diagnosis. In the investigation, we developed an extraction and derivatization protocol, using experimental design theory (design of experiment), for analyzing the human blood plasma metabolome by GC/MS. The protocol was optimized by evaluating the data for more than 500 resolved peaks using multivariate statistical tools including principal component analysis and partial least-squares projections to latent structures (PLS). The performance of five organic solvents (methanol, ethanol, acetonitrile, acetone, chloroform), singly and in combination, was investigated to optimize the LMC extraction. PLS analysis demonstrated that methanol extraction was particularly efficient and highly reproducible. The extraction and derivatization conditions were also optimized. Quantitative data for 32 endogenous compounds showed good precision and linearity. In addition, the determined amounts of eight selected compounds agreed well with analyses by independent methods in accredited laboratories, and most of the compounds could be detected at absolute levels of similar to 0.1 pmol injected, corresponding to plasma concentrations between 0.1 and 1 mu M. The results suggest that the method could be usefully integrated into metabolomic studies for various purposes, e.g., for identifying biological markers related to diseases.