Methods for the quantitation of human milk oligosaccharides in bacterial fermentation by mass spectrometry

Milady R. Ninonuevo, Robert E. Ward, Riccardo G. LoCascio, J. Bruce German, Samara L. Freeman, Mariana Barboza Gardner, David A. Mills, Carlito B Lebrilla

Research output: Contribution to journalArticlepeer-review

70 Scopus citations


Oligosaccharides are the third most abundant component in human milk. In the past decades, it became apparent that they would be able to protect against pathogens and participate in the development of the gut microflora for infants. However, their role in infants' nutrition and development remains poorly understood. To better understand this function, it is extremely important to have a quantitative tool for profiling oligosaccharides. In this article, we show the development of a method to quantitatively differentiate the relative amounts of oligosaccharides fermented by different intestinal bacteria. To determine the oligosaccharide consumption, bacteria were grown in a medium using human milk oligosaccharides (HMOs) as the only carbon source purified from breast milk and further analyzed by matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR MS). A method using an internal deuterium-labeled standard was developed and compared with an external standard method, with the internal standard method giving better precision and unambiguous measurements than the external standard method and providing to be a novel and robust tool for following bacterial fermentation of milk oligosaccharides.

Original languageEnglish (US)
Pages (from-to)15-23
Number of pages9
JournalAnalytical Biochemistry
Issue number1
StatePublished - Feb 1 2007


  • Bacterial fermentation
  • Human milk oligosaccharides
  • Prebiotics
  • Quantitation

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology


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