title: Development and performance of a gas chromatography-time-of-flight mass spectrometry analysis for large-scale nontargeted metabolomic studies of human serum.
abstract: 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/).
authors.affiliation: Bioanalytical Sciences Group, School of Chemistry, Manchester Interdisciplinary Biocentre, University of Manchester, M1 7DN, UK. paul.begley@manchester.ac.uk
authors.forename: Paul
authors.initials: P
authors.lastname: Begley
authors.forename: Sue
authors.initials: S
authors.lastname: Francis-McIntyre
authors.forename: Warwick B
authors.initials: WB
authors.lastname: Dunn
authors.forename: David I
authors.initials: DI
authors.lastname: Broadhurst
authors.forename: Antony
authors.initials: A
authors.lastname: Halsall
authors.forename: Andy
authors.initials: A
authors.lastname: Tseng
authors.forename: Joshua
authors.initials: J
authors.lastname: Knowles
authors.forename: Royston
authors.initials: R
authors.lastname: Goodacre
authors.forename: Douglas B
authors.initials: DB
authors.lastname: Kell
mesh_terms: D001798:Blood Proteins / Q000032:analysis; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D055432:Metabolomics; D011786:Quality Control; D015203:Reproducibility of Results
chemical_list: D001798:Blood Proteins