Structural characterization of fucose-containing oligosaccharides by high-performance liquid chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Citation
M. Suzuki et A. Suzuki, Structural characterization of fucose-containing oligosaccharides by high-performance liquid chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, BIOL CHEM, 382(2), 2001, pp. 251-257
Citations number
29
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOLOGICAL CHEMISTRY
ISSN journal
1431-6730 → ACNP
Volume
382
Issue
2
Year of publication
2001
Pages
251 - 257
Database
ISI
SICI code
1431-6730(200102)382:2<251:SCOFOB>2.0.ZU;2-V
Abstract
Eight pyridylamino (PA) derivatives of fucose-containing oligosaccharides, which occur as free oligosaccharides in human milk and also are derived fro m glycosphingolipids, have been analyzed by high-performance liquid chromat ography (HPLC) on normal-phase and reversed-phase columns, and by matrix-as sisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrom etry, Six out of eight PA-oligosaccharides were clearly separated by both n ormal- and reversed-phase HPLC at a column temperature of 40 degreesC, but two PA-oligosaccharides, lacto-N-fucopentaose II [Gal beta1-3(Fuc alpha1-4) GlcNAc beta1-3Gal beta1-4GlcPA] and lacto-N-fucopentaose III [Gal beta1-4(F uc alpha1-3)GlcNAc beta1-3Gal beta1-4GlcPA], were not separated. The two un resolved PA-oligosaccharides were finally separated by reversed-phase HPLC at a column temperature of 11 degreesC, MALDI-TOF mass spectra of PA-oligosaccharides demonstrated pseudo-molecular ions as the predominant signals, therefore information about the molecular mass of each PA-oligosaccharide was easily obtained. Post-source decay (PS D) MALDI-TOF mass spectra of PA-oligosaccharides gave information about the carbohydrate sequences and carbohydrate species of each PA-oligosaccharide by detecting the ions responsible for the cleavage of the glycosidic bonds . The detection limits of the PA-oligosaccharides by HPLC, MALDI-TOF mass s pectrometry, and PSD MALDI-TOF mass spectrometry were 20 fmol, 20 fmol, and 2 pmol, respectively, These results suggest that a system including HPLC a nd MALDI-TOF mass spectrometry or HPLC and PSD MALDI-TOF mass spectrometry is quite useful for the structural characterization of sub-pmol or pmol lev els of fucose-containing oligosaccharides, and that these methods could be used for the analysis of Various types of oligosaccharides derived from gly coproteins and glycosphingolipids.