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1

Department of Biotechnology, University of Rijeka, Radmile Matejčić 2, Rijeka 51000, Croatia

2

Waters Corporation, Stamford Avenue, Altrincham Road, Wilmslow SK9 4AX, UK

3

Institute for Hygiene, University of Muenster, Robert-Koch-Strasse 41, Muenster D-48149, Germany

4

Institute for medical Physics and Biophysics, University of Muenster, Robert-Koch-Strasse 31, Muenster D-48149, Germany





*

Author to whom correspondence should be addressed.



Abstract Human milk oligosaccharides HMO represent the bioactive components of human milk, influencing the infant’s gastrointestinal microflora and immune system. Structurally, they represent a highly complex class of analyte, where the main core oligosaccharide structures are built from galactose and N-acetylglucosamine, linked by 1-3 or 1-4 glycosidic linkages and potentially modified with fucose and sialic acid residues. The core structures can be linear or branched. Additional structural complexity in samples can be induced by endogenous exoglycosidase activity or chemical procedures during the sample preparation. Here, we show that using matrix-assisted laser desorption-ionization MALDI quadrupole-time-of-flight Q-TOF collision-induced dissociation CID as a fast screening method, diagnostic structural information about single oligosaccharide components present in a complex mixture can be obtained. According to sequencing data on 14 out of 22 parent ions detected in a single high molecular weight oligosaccharide chromatographic fraction, 20 different oligosaccharide structure types, corresponding to over 30 isomeric oligosaccharide structures and over 100 possible HMO isomers when biosynthetic linkage variations were taken into account, were postulated. For MS-MS data analysis, we used the de novo sequencing approach using diagnostic ion analysis on reduced oligosaccharides by following known biosynthetic rules. Using this approach, de novo characterization has been achieved also for the structures, which could not have been predicted. View Full-Text

Keywords: human milk oligosaccharides; MALDI Q-TOF matrix-assisted laser desorption-ionization quadrupole-time-of-flight MS mass spectrometry; CID collision-induced dissociation; diagnostic ion MS; de novo sequencing human milk oligosaccharides; MALDI Q-TOF matrix-assisted laser desorption-ionization quadrupole-time-of-flight MS mass spectrometry; CID collision-induced dissociation; diagnostic ion MS; de novo sequencing





Autor: Marko Jovanović 1,* , Richard Tyldesley-Worster 2, Gottfried Pohlentz 3 and Jasna Peter-Katalinić 1,4

Fuente: http://mdpi.com/



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