Development and Performance of a Gas Chromatography−Time-of-Flight Mass Spectrometry Analysis for Large-Scale Nontargeted Metabolomic Studies of Human Serum

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• 作者：Paul Begley ; Sue Francis-McIntyre ; Warwick B. Dunn ; David I. Broadhurst ; Antony Halsall ; Andy Tseng ; Joshua Knowles ; ; Royston Goodacre ; Douglas B. Kell
• 刊名：Analytical Chemistry
• 出版年：2009
• 出版时间：August 15, 2009
• 年：2009
• 卷：81
• 期：16
• 页码：7038-7046
• 全文大小：208K
• 年卷期：v.81,no.16(August 15, 2009)
• ISSN：1520-6882

文摘

A method for the preparation and GC−TOF-MS analysis of human serum samples has been developed and evaluated for application in long-term metabolomic studies. Serum samples were deproteinized using 3:1 methanol/serum, dried in a vacuum concentrator, and chemically derivatized in a two-stage process. Samples were analyzed by GC−TOF-MS with a 25 min analysis time. In addition, quality control (QC) samples were used to quantify process variability. Optimization of chemical derivatization was performed. Products were found to be stable for 30 h after derivatization. An assessment of within-day repeatability and within-week reproducibility demonstrates that excellent performance is observed with our developed method. Analyses were consistent over a 5 month period. Additional method testing, using spiked serum samples, showed the ability to define metabolite differences between samples from a population and samples spiked with metabolites standards. This methodology allows the continuous acquisition and application of data acquired over many months in long-term metabolomic studies, including the HUSERMET project (http://www.husermet.org/).