Neutral, charged and stratified polar monoliths for hydrophilic interaction capillary electrochromatography

• Various monoliths were successfully introduced and evaluated in HI-CEC.
• A hydroxy monolith based on diol functions yielded cathodal EOF.
• A cationic monolith based on primary amine functions (AP-monolith).
• Three sugar modified monoliths (SMAP-monoliths) called stratified monoliths blending the amine and hydroxy monoliths.
• The polarity of the various monoliths followed the order: hydroxy monolith < AP-monolith < SMAP-monoliths.

Novel polar monoliths were introduced for hydrophilic interaction capillary electrochromatography (HI-CEC). In one case, a neutral polar monolith resulted from the in situ polymerization of glyceryl methacrylate (GMM) and pentaerythritol triacrylate (PETA) in a ternary porogenic solvent. GMM and PETA possess hydroxyl functional groups, which impart the monolith with hydrophilic interaction sites. This monolith is designated as hydroxy monolith. Although the hydroxy monolith is neutral and void of fixed charges on the surface, a relatively strong cathodal EOF was observed due to the electric double layer formed by the adsorption of ions from the mobile phase, producing a bulk mobile phase flow. The second monolith is charged and referred to as AP-monolith that possesses amine/amide functionalities on its surface, and was prepared by the in situ polymerization of N-(3-aminopropyl) methacrylamide hydrochloride (NAPM) and ethylene dimethacrylate (EDMA) in the presence of cyclohexanol, dodecanol and methanol as porogens. Over the pH range studied a strong anodal EOF was observed. The AP-monolith was further exploited in HI-CEC by modifying its surface with neutral mono- and oligosaccharides to produce a series of the so called sugar modified AP-monoliths (SMAP-monolith), which are considered as stratified hydrophilic monoliths possessing a sub-layer of polar amine/amide groups and a top layer of sugar (a polyhydroxy top layer). The SMAP-monoliths can be viewed as a blend of both the hydroxy monolith and the AP-monolith. The polarity of the various monoliths seems to follow the order: hydroxy monolith < AP-monolith < SMAP-monolith. The novel monoliths were characterized over a wide range of elution conditions with a variety of polar solutes including phenols, substituted phenols, nucleic acid bases, nucleosides and nucleotides.