Optimising Core-Shell UHPLC Columns for Improving Protein and Peptide Separations

The introduction by several chromatography column manufacturers of sub-2 µm fully-porous as well as core-shell sub-3 µm UHPLC columns over the last several years have profoundly impacted the chromatography industry in the analysis of small molecules. These advances in silica particle technologies have for the most part focused on using increased efficiency as a means of reducing run times while maintaining resolution of existing separations.1,2 The fundamental concept for all new silica media (sub-2 µm or core-shell) has been to reduce analyte diffusion leading to sharper peaks. By narrowing particle size distribution and optimising packing band broadening effects due to eddy diffusion (the so-called ’A‘ term of the Van Deemter curve) are minimised leading to an overall increase in performance independent of mobile phase velocity.