Itaru Yazawa and Bryan Evans, Imtakt USA

Porous ODS columns are the most popular reversed-phase columns used in today's laboratories. Unfortunately, there are several shortcomings for porous ODS columns for polymer applications, including: poor peak shape of solutes due to wide range in pore size distribution; poor recovery of solutes due to micro-pores and meso-pores; and reduced column efficiency due to high mass transfer resistance. We have developed a 2 μm nonporous high resolution ODS column, Presto FF-C18, which can overcome these shortcomings for polymer separation.

Figure 1

Experimental and Results

All data was generated with semi-micro HPLC system equipped with UV or ELS detection. Figure 1 shows peptide separations (try ptic diges t of α-cas ein). Twice as many peaks were generated on Presto FF-C18 (2 μm nonporous) than with the 3 μm porous ODS column. Due to the nature of nonporous material (no diffusion in pores), slower flow rates can be used, which result in an increase in resolution and sensitivity. Figure 2 shows Immunoglobulin G (IgG) antibody analysis. Presto FF-C18 can provide improved peak shape and recognition between mAb (homogenous) and polyclonal antibody (heterogeneous). Figure 3 shows Hyaluronic acid analysis. Using gradient elution, Presto FF-C18 shows improved peak shape over SEC columns for polysaccharide analysis.

Figure 2

Conclusion

Presto FF-C18 shows improved resolution, peak shape, and sensitivity for polymer analysis. Presto FF-C18 can expand the separation possibilities when complimented with porous ODS columns.

Figure 3

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