A strategy for annotating the human milk glycome

Milady R. Ninonuevo, Youmie Park, Hongfeng Yin, Jinhua Zhang, Robert E. Ward, Brian H. Clowers, J. Bruce German, Samara L. Freeman, Kevin Killeen, Rudolf Grimm, Carlito B Lebrilla

Research output: Contribution to journalArticlepeer-review

356 Scopus citations


Oligosaccharides in human milk represent a group of bioactive molecules that have evolved to be an abundant and diverse component of human milk, even though they have no direct nutritive value to the infant. A recent hypothesis proposes that they could be substrates for the development of the intestinal microflora and the mucosal immune system. The inability to determine the exact composition of these oligosaccharides limits research and the ability to understand their biological functions. Oligosaccharides isolated from the lipids and proteins of individual human milk samples were analyzed by a combination of techniques including microchip liquid chromatography mass spectrometry (HPLC-Chip/MS) and matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FT ICR MS). Accurate mass measurements obtained using an orthogonal time-of-flight (o-TOF) mass spectrometry provided oligosaccharide composition for approximately 200 individual molecular species. Comparison of HPLC-Chip/MS profiles from five different women revealed variations in milk oligosaccharide compositions. HPLC-Chip/MS profiling provides a method for routinely identifying milk oligosaccharides. Tandem MS in combination with exoglycosidase digestion provides unambiguous differentiation of structural isomers.

Original languageEnglish (US)
Pages (from-to)7471-7480
Number of pages10
JournalJournal of Agricultural and Food Chemistry
Issue number20
StatePublished - Oct 4 2006


  • Exoglycosidases
  • HPLC-Chip/MS technology
  • Human milk oligosaccharides
  • Infrared multiphoton dissociation
  • Matrix-assisted laser desorption/ionization Fourier transform mass spectrometry
  • Milk glycome

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Food Science
  • Chemistry (miscellaneous)


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