Development of a gas chromatography/mass spectrometry based metabolomics protocol by means of statistical experimental design
Development of a gas chromatography/mass spectrometry based metabolomics protocol by means of statistical experimental design
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DOI：
10.1007/s11306-011-0283-6
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发表时间：
2012-02-01
期刊：
影响因子：
3.6
通讯作者：
Spegel, Peter
中科院分区：
文献类型：
--
作者：
Danielsson, Anders P. H.;Moritz, Thomas;Spegel, Peter
代谢组学是一个不断发展的研究领域，新的协议正在迅速发展，新的应用程序被发现。常见的应用包括生物标志物的发现和药物代谢的阐明。然而，这种方案的开发很少包括系统优化，然后用真实的样品进行验证。在这里，GC/MS为基础的协议，使用甲氧基化，然后与N-叔丁基二甲基甲硅烷基-N-甲基三氟乙酰胺（MTBSTFA）的甲硅烷基化的血浆代谢物的分析是彻底开发和优化从衍生化到检测与统计实验设计（DOE）。用血浆样品进行验证，证明该方法是有效的，快速和可靠的，在24小时内共进行了51次分析，线性响应，低检测限和良好的精密度。由于不存在葡萄糖过载，获得的色谱图更加清晰，并且发现MTBSTFA衍生化的数据漂移比MTBSTFA衍生化的数据漂移更小。
Metabolomics is a growing research field where new protocols are rapidly developed and new applications discovered. Common applications include biomarker discovery and elucidation of drug metabolism. However, the development of such protocols rarely includes a systematic optimization followed by validation with real samples. Here a GC/MS-based protocol using methoximation followed by silylation with N-tert-butyldi-methylsilyl-N-methyltrifluoroacetamide (MTBSTFA) for analysis of blood plasma metabolites is thoroughly developed and optimized from derivatization to detection with statistical design of experiments (DOE). Validation was performed with blood plasma samples and proved the methodology to be efficient, rapid and reliable with a total of 51 analyses performed in 24 h, with linear responses, low detection limits and good precision. The obtained chromatograms were much cleaner, due to the absence of glucose overloading, and the data was found to drift less with MTBSTFA derivatisation than with MTBSTFA derivatisation.