Polysaccharides liquid chromatography fractionation

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. 

The initial PIA purification method was developed by Mack et al. (3). These authors used a different, two-step chromatography protocol involving size-exclusion and ion exchange chromatography on Sephadex G-200, Q-Sepharose, and S-Sepharose. A similar purification method has been described recently to isolate a PIA-related polysaccharide polymer in E. coli (7). Briefly, E. coli cells were incubated in 50 raM Tris-HCL buffer (pH 8.0), 100 mg lysozyme, and 0.1 M EDTA at room temperature for 2 h. Phenol/chloroform extraction steps were performed to separate protein and debris contamination from the polysaccharide. Samples were concentrated by ultrafiltration devices (10,000 MW cut off) and fractionated on a fast protein liquid chromatography (FPLC) system with a Sephacryl S-2000 column (equilibration and elution buffer 0.1 MPBS, pH 7.4). 

High- performance liquid chromatography High resolution, different columns are available to separate polysaccharides with different properties Need initial fractionation (purification) by GFC or anion exchange methods [99-101]... 

Methods of Separation and Purification of Polysaccharides There are two main types of liquid chromatography techniques that are employed for the purification of polysaccharides, namely, ion-exchange chromatography and size-exclusion chromatography (SEC) [99]. Ion-exchange chromatography is widely used for the fractionation of polar and nonpolar polysaccharides. SEC is used for the purification of polysaccharides on the basis of molecular weights. 

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