Automated SPE-RP-HPLC fractionation of biofluids combined to off-line NMR spectroscopy for biomarker identification in metabonomics.
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.
Rezzi
Serge
S
BioAnalytical Science, Nestlé Research Center, PO Box 44, CH-1000 Lausanne 26, Switzerland. serge.rezzi@rdls.nestle.com
Vera
Francia Arce
FA
Martin
François-Pierre J
FP
Wang
Serena
S
Lawler
Dennis
D
Kochhar
Sunil
S
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*
D015415:Biomarkers