Chromatography oligosaccharide detection

The Lobry de Bruyn reaction can be carried out on the paper, followed by chromatography of the products.167 168 The procedure of Bayly and Bourne22 for forming benzylamine derivatives of oligosaccharides on filter paper has been mentioned previously. Enzymic hydrolysis of phosphate esters, in conjunction with paper chromatography, aids detection of the spots.126... 

Oligosaccharides were isolated preparatively by high-pH anion exchange chromatography carried out on a LC-system (Dionex Corporation, Sunnyvale CA) equipped with a CarboPac PA-1 column (9 x 250 mm), coupled to a Spectra System AS 3500 auto sampler. The detection was carried out using Pulsed Amperometric Detection (PAD-II). 

Hydrophilic interaction chromatography on Asahipak NH2P or Excel-pak CHA-P44 with pulsed amperometric detection has been used to fractionate malto-oligosaccharides.266 The Asahipak NH2P is a polyvinyl alcohol support with a polyamine bonded phase, and the Excelpak is a sulfonated polystyrene in the Zn+2 form. Amine adsorption of sialic acid-containing oligosaccharides was performed on a Micropak AX-5 column (Varian) using acetonitrile-water-acetic acid-triethylamine.267... 

Hardy, M. R. and Townsend, R. R., Separation of positional isomers of oligosaccharides and glycopeptides by high-performance anion-exchange chromatography with pulsed amperometric detection, Proc. Natl. Acad. Sci. U.S.A., 85, 3289, 1988. 

Kotani, N. and Takasaki, S., Analysis of 2-aminobenzamide-labeled oligosaccharides by high-pH anion-exchange chromatography with fluorometric detection, Anal. Biochem., 264, 66, 1998. 

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). 

Before discussing the various derivatives that have been used, it should be remembered that oligosaccharides are often obtained by chromatography on paper or on cellulose columns, and this may cause them to become contaminated by xylan this may be eliminated by extraction of the crude fractions with hot ethanol.474 For similar reasons, glucose that is really extraneous may be detected in fractions separated by electrophoresis on cellulose acetate.475 Gas-liquid chromatography was used to show the presence in pituitary glyco-... 

Humbel R, Collart M (1975) Oligosaccharides in the urine of patients with glycoprotein storage diseases. I. Rapid detection by thin-layer chromatography. Clin Chim Acta 60 143-145... 

To date, all TOPCAT donors prepared in our laboratory were isolated by column chromatography as pure (3-anomers (see Scheme 3), and they exhibited much higher stability compared with glycosyl 2-pyridylcarbonates. They are often crystalline and they can be stored for extended periods without a detectable change. These features are extremely important for their synthetic applications as versatile glycosyl donors in oligosaccharide and glycoside synthesis. 

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