Separation of Peptides and Oligonucleotides Using a Monolithic Polymer Layer and Pressurized Planar Electrophoresis and Electrochromatography

Abstract

The rapid separation of mixtures of six peptides using porous polymer monolithic layers in electrophoresis and pressurized planar electrochromatography modes has been achieved. The separations in the former mode were performed on a generic hydrophobic poly(butyl methacrylate-co-ethylene dimethacrylate) layer with no ionizable functionalities and required 2 min. This layer also enabled the separation of three oligonucleotides. The separation in the pressurized planar electrochromatographic mode was carried out using a negatively charged layer prepared via cografting of 2-acrylamido-2-methyl-1-propanesulfonic acid and 2-hydroxyethyl methacrylate on top of the generic hydrophobic monolith and was completed in 1 min.

High performance stationary phases for planar chromatography

“…Electroosmosis is used to drive the mobile phase flow through the layer in pressurized planar electrochromatography and requires an aqueous buffer as a component of the mobile phase [23,[57][58][59][60][61][62]]. An electric field applied across the layer is used to control the mobile phase velocity which is independent of the solvent front migration distance.…”

“…Monolithic films of silica gel [13,81,82] or organic polymers [58,83,84] are thin, continuous biporous structures cast on a supporting surface. These structures do not require a binder to form a stable layer.…”

“…Porous polymer monoliths are simply prepared in a thin film format using photoinitiated polymerization in a simple mold consisting of two glass plates, one of which provides the support structure for the monolithic film, separated with Teflon strips defining the film thickness [58,83,84]. Monolithic films 50-200 m thick were prepared from poly(butylmethacyrlate-co-ethylene dimethacylate) monomers with other functional groups introduced by grafting to provide different retention mechanisms or a charged surface for pressurized planar electrochromatography (Section 2.4) [58].…”

“…Monolithic films 50-200 m thick were prepared from poly(butylmethacyrlate-co-ethylene dimethacylate) monomers with other functional groups introduced by grafting to provide different retention mechanisms or a charged surface for pressurized planar electrochromatography (Section 2.4) [58]. There are no details of the kinetic performance of these layers or of their pore structure.…”

“…There are no details of the kinetic performance of these layers or of their pore structure. They were used for the separation of simple protein, peptide, and dye mixtures by one-and two-dimensional thin-layer chromatography with capillary flow (except for [58]). Different mass spectrometric techniques were generally used for detection.…”