Establishing the analytical platforms for characterizing human milk oligosaccharides is important to fully assess their specific functionalities. The characterization of these biomolecules, however, is still considered challenging, owing to their overall complexity and diversity. Addressed here are the technical difficulties with an emphasis on the application of mass spectrometry to rapidly profile and quantify human milk oligosaccharides. Fundamental concepts and improvements in instrumentation and an overview of the biological functions and structures of these compounds are also discussed. Results reveal that small-chain oligosaccharides, evident in abundance in the early stage of lactation, are selectively consumed by specific stains of Bifidobacterium longum biovar, infantis.
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