Oligosaccharides analysis

The introduction of g.l.c. for oligosaccharide analysis constituted a major breakthrough in the field. In addition to strategies for accurate and sensitive quantitation of monosaccharide type (19,20), chiral procedures may be adopted for enantiomeric (d and l) determination (21). The sensitivity of h.p.a.e. chromatography with pulsed amperometric detection now provides an alternative to g.l.c. for oligosaccharide compositional analysis (22). 

Oligosaccharide analysis using CE method provides faster run time compared to the HPLC method. Figure 27 shows a comparison of the oligosaccharide analysis using the HPFC and the CE method. 

CE oligosaccharide analysis with APTS labeling has been used in biotech industry for many years and has demonstrated reproducibility. It is now routinely used in GMP environment for release of therapeutic proteins and antibodies. Figure 29 shows the reproducibility of the analysis. 

FIGURE 27 Comparison of the HPLC and the CE method for oligosaccharide analysis. Panel A analysis using the CE method. Panel B analysis using the HPLC method. 

Most of the direct and indirect (transport) interfaces described here use chemical ionization (c.i.) ion-sources, which are not well suited to such polar, non-volatile compounds as tri- and higher oligosaccharides. The thermospray interface, which can operate on an ion-evaporative mode, is capable of producing intact molecular ions from such nonvolatile, polar molecules and should be useful in oligosaccharide analysis. Molecules of this type, however, can also be easily analyzed by fast-atom-bombardment ionization, and use of this technique, coupled to direct liquid introduction and moving-belt interfaces, has been reported. The latter system has been applied to complex oligosaccharide analysis. ... 

Analytical quality is maintained by including at least one known pathological sample in each run. There is no universal quality control scheme for oligosaccharide analysis due to the lack of sufficient patient samples. 

Jiang, Y. and Cole, R.B. (2005) Oligosaccharide analysis using anion attachment in negative mode electrospray mass spectrometry. J. Am. Soc. Mass Spectrom., 16 (1), 60-70. 

C. Bruggink, M. Wuhrer, C.A.M. Koeleman, V. Barreto, Y. Liu, C. Pohl, A. Ingendoh, C.H. Hokke, A.M. Deelder, Oligosaccharide analysis by capillary-scale HPAEC with on-line ion-trapMS, J. Chromatogr. B, 829 (2005) 136. 

N.G. Karlsson, N.L. Wilson, H.-J. Wirth, P. Dawes, Negative ion GCC nano-LC-MS increases sensitivity for glycoprotein oligosaccharide analysis. Rapid Commun. Mass Spectrom., 18 (2004)2282. 

P. Groves, M. Rasmussen, D. Molero, E. Samain, F. J. Canada, H. Driguez, and J. Jimenez-Barbero, Diffusion ordered spectroscopy as a complement to size exclusion chromatography in oligosaccharide analysis, Glycobiology, 14 (2004) 451-456. 

U. Schagerlof, H. Schagerlof, D. Momcilovic, G. Brinkmalm, and F. Tjemeld, Endoglucanase sensitivity for substituents in methyl cellulose hydrolysis studied using MALDI-TOFMS for oligosaccharide analysis and stmctural analysis of enzyme active sites. Biomacromolecules, 8 (2007) 2358-2365. 

A. Hannemann and V. N. Reinhold, Oligosaccharide analysis by mass spectrometry, in Encyclopedia of Biological Chemistry, Elsevier, New York, 2004. 

Oligosaccharide analysis The technique is based on the detection of specific oligosaccharides that are not normally present in the juice but are introduced by the addition of sugars that have undergone inversion. Analysis relies upon liquid chromatographic detection. 

---