Development of a Single Column Method for the Separation of Lipid- and Protein-Derived Oligosaccharides

• DOI: 10.1021/pr800704t
• Publication Date: 2008-12-19T21:23:16Z
• Authors: David C. A. Neville; Raymond A. Dwek; Terry D. Butters
• Keywords: Oligosaccharide; Anthranilic acid

Fluorescent labeling of oligosaccharides with anthranilic acid (2-aminobenzoic acid; 2AA), or 2-aminobenzamide (2AB) permits the rapid, sensitive analysis structures present in cells and tissues. Normal-phase (NP)/hydrophilic interaction chromatography (HILIC) is commonly used to separate fluorophore-derivatized oligosaccharides. Column elution expressed as glucose units (GU) following calculation relative retention when compared an external oligomer standard. However, there significant overlap between sialylated neutral anion-exchange (NP-AE) HPLC can differing classes according number charged residues, but times GU cannot be calculated across entire gradient. We have overcome this difficulty by use a Dionex AS11 column that combines both hydrophilic chromatographies, termed HIAX, which enables values for carry other negatively groups. The same method may also employed 2AB fluorophore-labeled Additionally, eluants are columns. Therefore, HILIC- HIAX-separated performed using single system set eluents simple change.