title: Characterization of glycosphingolipids by supercritical fluid chromatography-mass spectrometry.
abstract: Gangliosides have been characterized by supercritical fluid chromatography-chemical ionization mass spectrometry (SFC-CIMS) as permethyl and pertrimethylsilyl derivatives, using carbon dioxide as the SFC mobile phase and CI reagent gas. Ganglioside classes and ceramide heterogeneity within each class are well resolved by SFC. Direct SFC-interfacing allows the analytical manipulations of single-ion monitoring, total-ion plots, background subtraction, library searches, and spectral reconstruction algorithms. Addition of ammonia to the CI ion chamber (NH3 as a CI reagent gas) yields abundant molecular-weight-related ions, (MH)+ and (MNH4)+ from analyte derivatives. Substitution of methanol for ammonia yields considerable parent-ion fragmentation, providing structural information on carbohydrate sequence, fatty acid, and sphingoid components. Under these latter conditions a unique alpha-cleavage fragment is observed which differentiates fatty acid from sphingosine heterogeneity. For ganglioside samples, the carboxyl group of neuraminyl residue(s) have been esterified with pentafluorobenzyl bromide and the products analyzed by negative ion chemical ionization MS. This modification improves chemical selectivity and greatly enhances detecting sensitivity. These "soft" ionization conditions provide abundant molecular-weight-related anions for collision-induced dissociation and subpicogram detection.
authors.affiliation: Department of Nutrition, Harvard School of Public Health, Boston, Massachusetts 02115.
authors.forename: M V
authors.initials: MV
authors.lastname: Merritt
authors.forename: D M
authors.initials: DM
authors.lastname: Sheeley
authors.forename: V N
authors.initials: VN
authors.lastname: Reinhold
mesh_terms: D000641:Ammonia; D000818:Animals; D001923:Brain Chemistry; D002245:Carbon Dioxide; D005464:Fluorobenzenes; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006028:Glycosphingolipids / Q000032:analysis*; D015394:Molecular Structure; D005466:Fluorocarbons
chemical_list: D000641:Ammonia; D002245:Carbon Dioxide; D005464:Fluorobenzenes; D006028:Glycosphingolipids; C025063:pentafluorobenzyl bromide; D005466:Fluorocarbons