Quantification of individual oligosaccharide compounds from human milk using high-pH anion-exchange chromatography

A quantification method suitable for determination of individual oligosaccharide compounds from human milk has been established. The crude milk oligosaccharide fraction was separated into acidic oligosaccharides, neutral oligosaccharides, and lactose by gel permeation chromatography. After this separation step neutral and acidic oligosaccharides were analyzed separately by high-pH anion-exchange chromatography with pulsed amperometric detection. The concentrations of 14 neutral oligosaccharides, of 6 acidic oligosaccharides, and of N-acetylneuraminic acid were determined on the internal standards stachyose and galacturonic acid, respectively. Thus, previously applied quantification methods for milk oligosaccharides based on gel permeation chromatography have been decisively improved.