Recovery of free oligosaccharides from derivatives labeled by reductive amination

Abstract

This study examined chemical regeneration of free oligosaccharides from their fluorescent derivatives prepared by reductive amination with various aromatic amines. Maltose derivatives of ethyl 4-aminobenzoate ( p-ABEE), 2-aminobenzonitrile ( o-ABN), 4-aminobenzonitrile ( p-ABN), 7-amino-4-methylcoumarin (AMC), 2-aminobenzoic acid ( o-ABA), 2-aminobenzamide ( o-ABAD), 2-aminopyridine (AP), and 8-aminonaphthalene-1,3,6-trisulfonate (ANTS) were incubated at 30 °C with an aqueous solution of hydrogen peroxide/acetic acid. Recoveries of maltose from p-ABEE, p-ABN, and AMC derivatives were fairly good and gave approximately 90% of maltose. Recoveries of maltose from its o-substituted aniline ( o-ABA, o-ABAD, and o-ABN) derivatives were 5–40%, but maltose was unrecoverable from AP and ANTS derivatives. Nevertheless, prior treatment of an AP derivative with cyanogen bromide enabled the regeneration of maltose in high yields. As an application, p-ABEE-labeled N-glycans from some glycoproteins separated on an amide column were identified by converting peak components to their AP derivatives via free saccharides and following mapping by reversed-phase chromatography.