Considerations in Selecting Reversed-Phase Columns for Peptide Separations

Bill Warren, Waters Corp, MA, USA

The ability to effectively separate, identify, and quantitate peptides at substantially different concentrations is important in applications ranging from the analyses of biotherapeutic proteins to proteomics and biomarker investigations.  As such, LC and LC/MS using reversed-phase chromatography is frequently the separation method of choice due to its ability to resolve species that may differ by only a few amino acids.

This technical seminar will highlight various physical and chemical considerations that influence peptide separations using reversed-phase chromatography.

The following variables important in peptide separations will be addressed:

- Peak Capacity

- Retentivity

- Selectivity

- Particle Technologies (Solid-Core vs. Porous)

- Particle Size

- Selection of trap / elute and analytical RP columns for proteomics investigations