Polysaccharide Characterization by Aqueous Size Exclusion Chromatography and Low-Angle Laser Light Scattering

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Abstract
Aqueous size exclusion chromatography coupled with on-line low-angle laser light scattering (SEC/LALLS) is a valuable analytical tool for characterization of polysaccharides and other important biopolymers. This work reviews the fundamental size separation mechanism of polymers chromatographed via SEC, the development of SEC/LALLS methods for characterization of eluted polymers, and applications of this technique to determine polysaccharide physical and chemical properties. Important nonsize exclusion effects encountered in aqueous SEC of polysaccharides are discussed and attributed to intramolecular and polymer-support interactions, as well as flow-related anomalies. The necessity of absolute molecular weight detection as a direct means of calibration is presented. Low-angle laser light scattering coupled to SEC provides a simple method of direct calibration and allows determination of polymer molecular weight and molecular weight distribution. Recent applications of SEC/LALLS to determine polysaccharide branching characteristics are detailed. The combined knowledge of molecular weight distributions and branching distributions provides insight into the molecular kinetic events of polysaccharide processing operations.