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