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 journalArticle

59 Citations (Scopus)

Abstract

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
Volume361
Issue number1
DOIs
StatePublished - Feb 1 2007

Fingerprint

Human Milk
Oligosaccharides
Fermentation
Mass spectrometry
Mass Spectrometry
Bacteria
Cyclotrons
Cyclotron resonance
Deuterium
Pathogens
Fourier Analysis
Nutrition
Child Development
Ionization
Desorption
Fourier transforms
Milk
Lasers
Carbon
Ions

Keywords

  • Bacterial fermentation
  • Human milk oligosaccharides
  • MALDI-FTICR MS
  • Prebiotics
  • Quantitation

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Methods for the quantitation of human milk oligosaccharides in bacterial fermentation by mass spectrometry. / Ninonuevo, Milady R.; Ward, Robert E.; LoCascio, Riccardo G.; German, J. Bruce; Freeman, Samara L.; Barboza Gardner, Mariana; Mills, David A.; Lebrilla, Carlito B.

In: Analytical Biochemistry, Vol. 361, No. 1, 01.02.2007, p. 15-23.

Research output: Contribution to journalArticle

Ninonuevo, MR, Ward, RE, LoCascio, RG, German, JB, Freeman, SL, Barboza Gardner, M, Mills, DA & Lebrilla, CB 2007, 'Methods for the quantitation of human milk oligosaccharides in bacterial fermentation by mass spectrometry', Analytical Biochemistry, vol. 361, no. 1, pp. 15-23. https://doi.org/10.1016/j.ab.2006.11.010
Ninonuevo, Milady R. ; Ward, Robert E. ; LoCascio, Riccardo G. ; German, J. Bruce ; Freeman, Samara L. ; Barboza Gardner, Mariana ; Mills, David A. ; Lebrilla, Carlito B. / Methods for the quantitation of human milk oligosaccharides in bacterial fermentation by mass spectrometry. In: Analytical Biochemistry. 2007 ; Vol. 361, No. 1. pp. 15-23.
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