title: Femtomole oligosaccharide detection using a reducing-end derivative and chemical ionization mass spectrometry.
abstract: A bimodal reagent (pentafluorobenzyl aminobenzoate) has been synthesized to improve oligosaccharide isolation, detection, and structural characterization. The reagent is glycosidically attached to the reducing end of glycan residues, imparts fluorescent and uv properties for chromatographic detection, and functions as an efficient electron trap under negative ion chemical ionization mass spectrometry for femtomole detectability. Facile ester cleavage and pentafluorobenzyl elimination provides a single molecular-weight-related fragment in high abundance. Procedures are described for reagent synthesis, purification, and oligosaccharide conjugation. Carbohydrate samples derivatized with this reagent are evaluated by high-performance liquid chromatography and supercritical fluid chromatography (SFC) and for sensitivity by SFC negative ion chemical ionization mass spectrometry.
authors.affiliation: Harvard University School of Public Health, Boston, Massachusetts 02115.
authors.forename: J P
authors.initials: JP
authors.lastname: Caesar
authors.forename: D M
authors.initials: DM
authors.lastname: Sheeley
authors.forename: V N
authors.initials: VN
authors.lastname: Reinhold
mesh_terms: D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents; D006031:Glycosylation; D013058:Mass Spectrometry; D008722:Methods; D015394:Molecular Structure; D008970:Molecular Weight; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification*; D062366:para-Aminobenzoates; D005464:Fluorobenzenes; D005466:Fluorocarbons
chemical_list: D003432:Cross-Linking Reagents; D009844:Oligosaccharides; D062366:para-Aminobenzoates; C068258:pentafluorobenzyl-4-aminobenzoic acid; D005464:Fluorobenzenes; D005466:Fluorocarbons