High-pH anion-exchange chromatography with pulsed amperometric detection and molar response factors of human milk oligosaccharides

A method is described to separate and characterize neutral and acidic lactose-derived oligosaccharides without prior derivatization or reduction by high-pH anion-exchange chromatography and pulsed amperometric detection (HPAEC-PAD). This method has been applied to human milk oligosaccharides from do...

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Bibliographic Details
Published in:Journal of chromatography. B, Biomedical applications Biomedical applications, 1996-10, Vol.685 (2), p.211-221
Main Authors: Kunz, Clemens, Rudloff, Silvia, Hintelmann, Annette, Pohlentz, Gottfried, Egge, Heinz
Format: Article
Language:English
Subjects:
Biological and medical sciences
Carbohydrate Sequence
Chromatography, Gel
Chromatography, Ion Exchange
Chromatography, Liquid - methods
Chromatography, Thin Layer
Female
Fundamental and applied biological sciences. Psychology
Human milk
Humans
Hydrogen-Ion Concentration
Lewis Blood-Group System
Milk, Human - chemistry
Mother. Fetoplacental unit. Mammary gland. Milk
Oligosaccharides
Oligosaccharides - analysis
Oligosaccharides - chemistry
Pregnancy. Parturition. Lactation
Vertebrates: reproduction
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Summary:A method is described to separate and characterize neutral and acidic lactose-derived oligosaccharides without prior derivatization or reduction by high-pH anion-exchange chromatography and pulsed amperometric detection (HPAEC-PAD). This method has been applied to human milk oligosaccharides from donors with different blood group specificity (A, Le a and A, Le b). Neutral and acidic components were separated from each other by anion-ecchange chromatography. A distinct separation of individual components was obtained by size-exclusion chromatography on Fractogel TSK HW 50S (acidic oligosaccharides) or Fractogel TSK HW 40S (neutral oligosaccharides containing up to 6 monomers) and Bio-Gel P-4 size exclusion (neutral oligosaccharides containing more than 6 monomers). Furthermore, the molar response factors after HPAEC-PAD have been determined for 8 components.
ISSN:0378-4347
1572-6495
DOI:10.1016/S0378-4347(96)00181-8