Ultrahigh-Efficiency LC-MS/MS Separations of Proteolytic Digests on PepSwift Monolithic Columns

This application note evaluates the LC performance of 50 mm, 250 mm, and 1 m capillary poly(styrene-co-divinylbenzene) monolithic columns for LC-MS/MS peptide-mapping experiments, and discusses the optimization of LC conditions (including flow rate and gradient time) and column length. When analyzing complex peptide mixtures, the identification and quantification of low abundance peptides by MS is often hindered by insufficient separation and ion-suppression effects. To improve the separation of complex proteomic samples, increasing effort is directed to the development of novel column technology. The robustness of these high-efficiency monolithic materials, covalently bonded to the capillary wall, and their excellent retention-time stability makes this column technology very attractive in comparison to packed capillary columns.