N-methyl-O-benzylhydroxylamine derivatives of sialylated oligosaccharides, their synthesis and separation with reversed-phase HPLC

• Published in: Carbohydrate research Vol. 533; p. 108939
• Main Authors: Norberg, Thomas, Kallin, Elisabet
• Format: Journal Article
• Language: English
• Published: London Elsevier Ltd 01.11.2023; Elsevier
• Subjects: Biochemistry & Molecular Biology; Chemistry; Chemistry, Applied; Chemistry, Organic; Derivatization; Hydroxylamines; Life Sciences & Biomedicine; Milk oligosaccharides; Physical Sciences; Reversed-phase chromatography; Science & Technology; Derivatization; Milk oligosaccharides; Reversed-phase chromatography; Hydroxylamines
• ISSN: 0008-6215; 1873-426X; 1873-426X
• DOI: 10.1016/j.carres.2023.108939

The human milk trisaccharide 3′- sialyllactose was reacted with an excess of N-methyl-O-benzylhydroxylamine (MBHA) and the product 3′- sialyllactose-MBHA was isolated in high (91%) yield by solid-phase extraction. The isomeric trisaccharide 6′-sialyllactose-MBHA was also prepared, in this case by enzymatic sialylation of lactose-MBHA. A 50/50 mixture of the two sialyllactose-MBHA derivatives was easily separated by reversed-phase HPLC. The free oligosaccharides were recovered from their respective MBHA derivatives by acid hydrolysis. [Display omitted] •Reducing end oligosaccharides were derivatized with N-methyl-O-benzylhydroxylamine (MBHA) to form MBHA-oligosaccharides.•A mixture of 3′- and 6′-sialyllactose-MBHA oligosaccharides was separated by reversed-phase HPLC.•Sialylated MBHA-oligosaccharides were isolated from enzymatic reaction mixtures by C-18 solid-phase extraction.•Sialylated MBHA-oligosaccharides were easily hydrolyzed back to reducing end oligosaccharides.