Browsing by Subject "rakenneanalyysi"

The aim of the present study was to implement a tandem mass spectrometry (MS/MS) method for determining glycosidic bond linkage positions of neutral isomeric glucooligosaccharides. The methods for structural analysis and the fragmentation mechanisms of oligosaccharides in MS/MS-analysis was reviewed. The MS/MS-spectra of oligosaccharide contains glycosidic bond and cross ring cleavage fragment ions. The linkage position can be determined by cross ring cleavage fragment ions of cationized or anionized oligosaccharides. The study started by analyzing the typical MS/MS-spectra of model isomeric disaccharides with different linkage positions. The data was then used to assess the MS/MS-spectra of model tri- and tetrasaccharides. The similarity of MS/MS fragmentation patterns of the model disaccharides and the tri- and tetrasaccharides would enable the linkage position analysis by the applied MS/MS method. The MS/MS-analyses were carried out by using electron spray ionization with ion trap mass spectrometer in both positive and negative mode. In positive mode the oligosaccharides were analyzed as sodium and lithium adduct ions while chloride adduct ions were used in negative mode analysis. The different linkage positions of disaccharides were characterized by the different MS/MS fragmentation profiles in positive and negative ionization methods. The reducing end linkage positions of tri- and tetrasaccharides were easily identified by comparing their MS/MS fragmentation patterns with the MS/MS-spectra of model disaccharides. The other linkages in the tri- and tetrasaccharide were identified by using negative mode. In positive mode the identification of other linkages was not possible because the cross ring cleavage fragment ions were formed mostly from the reducing end. The linkages of tri- and tetrasaccharides could not be identified by MS3-analysis due to formation of isomeric glycosidic bond fragment ions as the charge can be retained in the reducing or non-reducing end. The results indicated that the applied MS/MS method was suitable for determing the glycosidic bond linkages of oligosaccharides in negative mode. In positive mode only the reducing end linkage can be determined.