title: Automated SPE-RP-HPLC fractionation of biofluids combined to off-line NMR spectroscopy for biomarker identification in metabonomics.
abstract: NMR-based metabonomics is a valuable and straightforward approach to measuring hundreds of metabolites in complex biofluids. However, metabolite identification is sometimes limited by overlapped signals in NMR spectra. We describe a new methodology using an automated hyphenation of solid phase extraction (SPE) with RP-HPLC combined to NMR spectroscopy, which allowed identification of 72 metabolites of various molecular classes in human urine. This methodology was also successfully applied to the fractionation of a cat urine sample to aid identification of aromatic compounds and felinine. The SPE-RP-HPLC method appears to be a reliable tool to support biomarker discovery in metabonomic studies.
authors.affiliation: BioAnalytical Science, Nestlé Research Center, PO Box 44, CH-1000 Lausanne 26, Switzerland. serge.rezzi@rdls.nestle.com
authors.forename: Serge
authors.initials: S
authors.lastname: Rezzi
authors.forename: Francia Arce
authors.initials: FA
authors.lastname: Vera
authors.forename: François-Pierre J
authors.initials: FP
authors.lastname: Martin
authors.forename: Serena
authors.initials: S
authors.lastname: Wang
authors.forename: Dennis
authors.initials: D
authors.lastname: Lawler
authors.forename: Sunil
authors.initials: S
authors.lastname: Kochhar
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis*; D002415:Cats; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019295:Computational Biology / Q000379:methods*; D006801:Humans; D008297:Male; D008660:Metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009862:Online Systems; D052616:Solid Phase Extraction / Q000379:methods*; D014556:Urine / Q000737:chemistry*
chemical_list: D015415:Biomarkers