Reversed-Phase Analysis of Polysaccharides Using a 2-µm Non-Porous ODS Phase

Bryan Evans and Itaru Yazawa, Imtakt USA

Polysaccharides are often separated using SEC columns. However, SEC columns do have limitations, specifically: poor column efficiency or peak shape. An alternative to SEC (and CE) for polysaccharide analysis is reversed-phase mode, utilizing Presto FF-C18 (2 μm nonporous ODS).

Figure 1: Hyaluronic acid.

Experimental and Results

All data was generated with semi-micro HPLC system equipped with ELS detection. Figure 1 shows analysis for hyaluronic acid. Different retention times were obtained by adjusting the beginning or ending organic composition. Figure 2 shows analysis for an endotoxin (a lipopolysaccharide from E. Coli). Figure 3 shows analysis for dextrans (up to 40 MDa). Even though the molecular weight is extremely large, the retention time and peak shape is acceptable for quantification.

Figure 2: Lipopolysaccharide (from E. coli O127).

Figure 3: Dextrans.

Conclusion

Presto FF-C18 (2 μm nonporous ODS) provides an alternative to SEC columns for biopolymer separations.

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