Critical Parameters Affecting the Reproducibility of Reversed-Phase Columns for Peptide Separations
Column-to-column variability has been referred to as the Achilles Heel in the development of rugged and reproducible HPLC methods for protein pharmaceuticals. Numerous column choices are available today to the separation chemist with each manufacturer claiming to have the most reproducible product. How does one make the best choice? The critical factors affecting column-to-column reproducibility for small molecule separations have been the topic of numerous studies. These studies are typically performed using small molecular acidic, basic and neutral probes at various mobile phase pH''s. However, very little information has been reported correlating the data generated using these small molecular probes to column-to-column reproducibility for peptide and protein separations.