Natural polysaccharides exhibit a wide range of biological activities, which are closely related to their structural characteristics, including their molecular weight distribution, size, monosaccharide composition, glycosidic bond types and spatial conformation, etc. Size exclusion chromatography (SEC) and asymmetrical flow field-flow fractionation (AF4), as two potent separation techniques, both harbor potential for continuous development and enhancement. This manuscript reviewed the fundamental principles and separation applications of SEC and AF4. The structural information and spatial conformation of polysaccharides can be obtained using SEC or AF4 coupled with multiple detectors. In addition, this manuscript elaborates in detail on the shear degradation of samples such as polysaccharides separated by SEC. In addition, the abnormal elution that occurs during the application of the two methods is also discussed. Both SEC and AF4 possess considerable potential for ongoing development and refinement, thereby offering increased possibilities and opportunities for polysaccharide separation and characterization.
Keywords: AF4; Abnormal elution; Polysaccharide; SEC; Shear degradation; Structural characterization.
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