title: Gas chromatography/flame ionisation detection mass spectrometry for the detection of endogenous urine metabolites for metabonomic studies and its use as a complementary tool to nuclear magnetic resonance spectroscopy.
abstract: Metabonomics is a relatively new field of research in which the total pool of metabolites in body fluids or tissues from different patient groups is subjected to comparative analysis. Nuclear magnetic resonance (NMR) spectroscopy is the technology that is currently most widely used for the analysis of these highly complex metabolite mixtures, and hundreds of metabolites can be detected without any upfront separation. We have investigated in this study whether gas chromatography (GC) separation in combination with flame ionisation detection (FID) and mass spectrometry (MS) detection can be used for metabolite profiling from urine. We show that although GC sample preparation is much more involved than for NMR, hundreds of metabolites can reproducibly be detected and analysed by GC. We show that the data quality is sufficiently high--particularly if appropriate baseline correction and time-warping methods are applied--to allow for data comparison by chemometrics methods. A sample set of urines from eleven healthy human volunteers was analysed independently by GC and NMR, and subsequent chemometrics analysis of the two datasets showed some similar features. As judged by NIST database searches of the GC/MS data some of the major metabolites that are detected by NMR are also visible by GC/MS. Since in contrast to NMR every peak in GC corresponds to a single metabolite, the electron ionisation spectra can be used to quickly identify metabolites of interest if their reference spectra are present in a searchable database. In summary, we show that GC is a method that can be used as a complementary tool to NMR for metabolite profiling of urine samples.
authors.affiliation: Pfizer Global R&D, Ramsgate Road, Sandwich CT13 9NJ, UK.
authors.forename: Sally-Ann
authors.initials: SA
authors.lastname: Fancy
authors.forename: Olaf
authors.initials: O
authors.lastname: Beckonert
authors.forename: Gareth
authors.initials: G
authors.lastname: Darbon
authors.forename: Warren
authors.initials: W
authors.lastname: Yabsley
authors.forename: Rosalind
authors.initials: R
authors.lastname: Walley
authors.forename: David
authors.initials: D
authors.lastname: Baker
authors.forename: George L
authors.initials: GL
authors.lastname: Perkins
authors.forename: Frank S
authors.initials: FS
authors.lastname: Pullen
authors.forename: Klaus
authors.initials: K
authors.lastname: Rumpel
mesh_terms: D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008297:Male; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D016482:Urinalysis / Q000379:methods*
chemical_list: D020543:Proteome