Chromatography of polysaccharides

ANALYTICAL AND PREPARATIVE COLUMNS FOR AQUEOUS SIZE EXCLUSION CHROMATOGRAPHY OF POLYSACCHARIDES... 

Yu, L. -P, Rollangs, J. E. (1987). Low-angle laser light scattering-aqueous size exclusion chromatography of polysaccharides molecular weight distribution and polymer branching determination. Journal of Applied Polymer Science, 33, 1909-1921. 

Chromatography of polysaccharides on silica gel and on carbon columns has been used to a limited extent. The use of ion-exchange resins has been investigated by Deuel and coworkers. The colunms had low capacities, and part of the material appeared to be irreversibly adsorbed. Deuel and coworkers, however, have reported better results when columns of cationic derivatives of cellulose, for instance (diethylaminoethyl)-cellulose, are used. Electrophoresis both by the Tiselius method on columns and on filter paper or glass-fiber sheets can give good separa-... 

Gel chromatography on Sephadex GlOO (2.8x50cm) of polysaccharide fraction I (Sample 2). The polysaccharide fraction was dissolved in a phosphate buffer at pH 6. After centrifugation, the supernatant was applied to the column at a flow rate of 0.8 ml/min. The elution was performed with a phosphate buffer and fractions of 10 ml each were collected. The refraction of each fraction was measured interferometrically. Fractions with coincident peaks were collected and analyzed for content of galacrturonic acid, neutral sugars and protein. 

For many of the organic materials in seawater, some form of chemical pretreatment is necessary before analysis is possible. The obvious cases are the hydrolysis of polysaccharides and proteins before the analysis for monomeric constituents, and the formation of volatile derivatives to permit analysis by gas chromatography. These methods will be discussed further in Chap. 9. 

Matthijs, N., Perrin, C., Maftouh, M., Massart, D.L., Vander Heyden, Y. Definition and system implementation of strategies for method development of chiral separations in normal- or reversed-phase liquid chromatography using polysaccharide-based stationary phases. 7. Chromatogr. A 2004, 1041, 119-133. 

Stringham, R.W Chiral separation of amines in subcritical fluid chromatography using polysaccharide stationary phases and acidic additives. J. Chromatogr. A 2005, 1070, 163-170. 

Englyst, H.N. and Cummings, J.H. (1984) Simplified method for the measurement of total non-starch polysaccharides by gas-liquid chromatography of constituent sugars as alditol acetates. Analyst 109, 937-942. 

In addition to classical reverse phase separation of peptides on octadecyl derivatized silica monoliths, sugars and peptides as well as proteins and nucleosides have been analyzed on a 20-cm-long silica-based poly(acrylic acid) column (ID. 200 pm), employing HILIC and weak cation-exchange chromatography, respectively [194]. Furthermore, HILIC fractionation of polysaccharides delivered remarkable and promising results [84,194]. 

Cellulose is another example of polysaccharides which is used as support in affinity chromatography. Cellulose has a historical significance. Phospo- and DNA-cellulose are used especially for DNA related separations [14]. Antibody and enz)nne purifications have also been carried out. However its fibrous and non-rmiform character limits its use since cellulose detains macromolecules [11]. 

In examining the structure of a polysaccharide, it is convenient to consider the methods involved under the three main headings (a) quantitative analysis, (b) methylation, and (c) periodate oxidation. These techniques may be supplemented by partial or enzymic hydrolysis as the circumstances indicate. Each of these aspects of polysaccharide chemistry may be aided by the application of gas-liquid chromatography, either qualitative or quantitative, or both. Thus, separations impossible by other techniques may often be achieved, and analytical data obtained in a fraction of the time demanded by other methods. 

In an attempt to make this account as complete as possible, certain related aspects, such as the hydrolysis of polysaccharides, are briefly discussed, but, in general, the only references cited are to those papers that have also mentioned use of gas-liquid chromatography. These ancillary sections must, therefore, not be considered exhaustive in treatment, but rather to be representative. 

Although this article is primarily concerned with gas-liquid chromatography of simple carbohydrates, it is worth noting that certain polysaccharides have been trimethylsilylated. Thus, chlorotrimethyl-silane in pyridine yielded fully trimethylsilylated amylose, cellulose, and pullulan, but failed with dextran and amylopectin.196 Cellulose, amylose, and polyvinyl alcohol have also been trimethylsilylated in molten N-(trimethylsilyl)acetamide.197 As examples of a new class of... 

---