Negative Ion Fast-Atom Bombardment Tandem Mass Spectrometry To Determine Sulfate and Linkage Position in Glycosaminoglycan-Derived Disaccharides

Abstract

Negative ion fast-atom bombardment tandem mass spectrometry has been used in the analysis of monosulfated. disaccharides. These commercially obtained disaccharides have been enzymatically prepared from glycosaminoglycans using polysaccharide lyases. Three disaccharides from chondroitin sulfate and dermatan sulfate and two disaccharides from heparan sulfate and chemically derivatized heparin were analyzed. All five disaccharides were isomeric, with differences in sulfate position and linkage position. The full-scan mass spectra are useful in differentiating isomers when the sulfate group resides on different saccharide units. This structural information was obtained from fragment ions produced through cleavage at the glycosidic linkage. The full-scan mass spectra of each monosulfated disaccharide also produced intense molecular anions having long lifetimes. Collisional activation of these resulted in tandem mass spectra rich in significant product ions. Some of these fragment ions were formed through ring cleavage and were useful in the determination of both sulfate and linkage position.

Tandem mass spectrometric strategies for determination of sulfation positions and uronic acid epimerization in chondroitin sulfate oligosaccharides

“…Intact molecular ions could, however, be produced using electrospray ionization (ESI) in the negative mode, and tandem MS has been applied to the CS oligosaccharide ions so generated [23,24]. Because CS disaccharides sulfated at the 4-or 6-position of the GalNAc residue, respectively, produce distinct CID product ion patterns [25][26][27], quantitative measurements based on ion abundances provide useful information on the global domain organization of CS chains [28]. It has also been shown that the information content of tandem mass spectra generated from CS oligosaccharides is strongly dependent on the charge state of the precursor ion [24].…”

Structure Elucidation of Hexasaccharide Derivatives Obtained from Keratan Sulfates by Ion Trap Mass Spectrometry.

“…1)῍11) For example, fast atom bombardment ionization followed by collisioninduced dissociation tandem mass spectrometry ῌFAB-CID-(MS) 2 ῍ has been applied to the characterization of glycosaminoglycan-derived oligosaccharides. 12), 13) We have demonstrated that FAB-CID-(MS) 2 can be successfully applied to the characterization of several oligosaccharides originating from chondroitin sulfates, 14), 15) hyaluronan, 15) heparin and heparan sulfates, 16), 17) tetrasaccharide tri-and tetra-sulfates from immunoreactive chondroitin sulfate fragments, 18) and oligosaccharides of the lactosamine series derived from keratan sulfates. 19) In this paper we would like to demonstrate the structure elucidation of a modified hexasaccharide obtained from keratan sulfates by the ESI-ion trap (MS) n technique.…”

ESI-MS differential fragmentation of positional isomers of sulfated oligosaccharides derived from carrageenans and agarans

“…In this context, a high number of publications have been focused on the study of sulfated sugar-containing macromolecules through the mass spectrometric analysis of their enzymatically or chemically produced oligosaccharides [8 -12]. The earlier studies [13,14] in this field were mostly conducted using FAB ionization. More recently, a number of combinations of ionization techniques (MALDI [15][16][17] and ESI [18 -22] and mass analyzers (TOF [15][16][17]22] quadrupoles and/or ion traps [8 -12, 18 -22] have been utilized.…”

“…In this context, a high number of publications have been focused on the study of sulfated sugar-containing macromolecules through the mass spectrometric analysis of their enzymatically or chemically produced oligosaccharides [8 -12]. The earlier studies [13,14] [8,9], and mucopolysaccharides [24] have served as models for the improvement of the knowledge of mass spectrometric methods involving these types of molecules.Many of the aforementioned studies have been dedicated to determine the sulfation position of isomeric oligosaccharides. Evaluation of the losses of monosaccharide units by sequencing mass spectrometry is normally sufficient to determine the sulfation pattern of isomers bearing sulfate groups on different monosaccharide units [10,17].…”

“…In this context, a high number of publications have been focused on the study of sulfated sugar-containing macromolecules through the mass spectrometric analysis of their enzymatically or chemically produced oligosaccharides [8 -12]. The earlier studies [13,14] [8,9], and mucopolysaccharides [24] have served as models for the improvement of the knowledge of mass spectrometric methods involving these types of molecules.…”