Automated SPE-RP-HPLC fractionation of biofluids combined to off-line NMR spectroscopy for biomarker identification in metabonomics

J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Aug 15;871(2):271-8. doi: 10.1016/j.jchromb.2008.04.001. Epub 2008 Apr 8.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / analysis*
  • Cats
  • Chromatography, High Pressure Liquid / methods*
  • Computational Biology / methods*
  • Humans
  • Male
  • Metabolism*
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Online Systems
  • Solid Phase Extraction / methods*
  • Urine / chemistry*

Substances

  • Biomarkers