Monosaccharide composition analysis

The quality of this library was examined by monosaccharide compositional analysis. A portion of the trisaccharide library was treated with aqueous trifluroacetic acid at 100°C for 4 h. Analysis of the resulting mixture of monosaccharides on a Dionex HPLC system with a PA1 column and pulsed amperometric detection (PAD) showed that galactose, glucose, and mannose were present in approximately the required ratio. 

Monosaccharide composition analysis of the intact MAb revealed monosaccharide ratios consistent with the presence of lactosamine type, fucosylated biantennaiy oligosaccharides with less than complete galactosylation (gal man is < 2 3) (Table 1). 

Monosaccharide composition analysis is often done to detect the presence of uncommon or atypical monosaccharides that may be present on the protein backbone, often in O- or C-linked forms. Typically, monosaccharides are released in strong acid at high temperature. However, it must be noted that hexose linkages are usually more stable to heat and strong acid than those of fucose or sialic acids. For instance, sialic acids are known to decompose after prolonged exposure to heat and acid. Optimizing the release of monosaccharides require careful sample... 

The monosaccharide composition of all the samples was determined by GLC analysis of the trimethylsilyl methyl glycoside derivatives [1]. 

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). 

For monitoring the extent of polysaccharide hydrolysis, l.c. methods that sepeu ate and analyze the non-fermentable oligosaccharides (d.p. 3-30) derived from cellulose, hemicellulose, and pectins are useful, and have already been described (see Section III,l,c). For determination of the monosaccharide composition of completely hydrolyzed, plant polysaccharides, l.c. is especially useful and has been applied to the compositional analysis of hydrolyzed plant fiber,wood pulps,plant cell-walls,and cotton fibers.In these representative examples, the major sugars of interest, namely, glucose, xylose, galactose, arabinose, and mannose, have traditionally been difficult to resolve by l.c. The separa-... 

Today (2000), GC/MS is still the method of choice for the determination of the monosaccharide composition of oligosaccharides and is widely used. The analysis starts with the hydrolysis or alcoholysis of the oligosaccharide to monosaccharides, which after derivatization (mostly by trimethylsilylation) are analysed by GC/MS. [221] Monosaccharide identification is based on comparison of retention time and fragmentation pattern with references. 

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