title: Towards high-throughput metabolomics using ultrahigh-field Fourier transform ion cyclotron resonance mass spectrometry.
abstract: With unmatched mass resolution, mass accuracy, and exceptional detection sensitivity, Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FTICR-MS) has the potential to be a powerful new technique for high-throughput metabolomic analysis. In this study, we examine the properties of an ultrahigh-field 12-Tesla (12T) FTICR-MS for the identification and absolute quantitation of human plasma metabolites, and for the untargeted metabolic fingerprinting of inbred-strain mouse serum by direct infusion (DI). Using internal mass calibration (mass error ≤1 ppm), we determined the rational elemental compositions (incorporating unlimited C, H, N and O, and a maximum of two S, three P, two Na, and one K per formula) of approximately 250 out of 570 metabolite features detected in a 3-min infusion analysis of aqueous extract of human plasma, and were able to identify more than 100 metabolites. Using isotopically-labeled internal standards, we were able to obtain excellent calibration curves for the absolute quantitation of choline with sub-pmol sensitivity, using 500 times less sample than previous LC/MS analyses. Under optimized serum dilution conditions, chemical compounds spiked into mouse serum as metabolite mimics showed a linear response over a 600-fold concentration range. DI/FTICR-MS analysis of serum from 26 mice from 2 inbred strains, with and without acute trichloroethylene (TCE) treatment, gave a relative standard deviation (RSD) of 4.5%. Finally, we extended this method to the metabolomic fingerprinting of serum samples from 49 mice from 5 inbred strains involved in an acute alcohol toxicity study, using both positive and negative electrospray ionization (ESI). Using these samples, we demonstrated the utility of this method for high-throughput metabolomics, with more than 400 metabolites profiled in only 24 h. Our experiments demonstrate that DI/FTICR-MS is well-suited for high-throughput metabolomic analysis.
authors.affiliation: J. Han, C. H. Borchers, University of Victoria - Genome BC Proteomics Centre, Vancouver Island Technology Park, #3101-4464 Markham St., Victoria, BC, Canada V8Z 7X8.
authors.forename: Jun
authors.initials: J
authors.lastname: Han
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Danell
authors.forename: Jayanti R
authors.initials: JR
authors.lastname: Patel
authors.forename: Dmitry R
authors.initials: DR
authors.lastname: Gumerov
authors.forename: Cameron O
authors.initials: CO
authors.lastname: Scarlett
authors.forename: J Paul
authors.initials: JP
authors.lastname: Speir
authors.forename: Carol E
authors.initials: CE
authors.lastname: Parker
authors.forename: Ivan
authors.initials: I
authors.lastname: Rusyn
authors.forename: Steven
authors.initials: S
authors.lastname: Zeisel
authors.forename: Christoph H
authors.initials: CH
authors.lastname: Borchers