Comparison of newly developed hydroxyl-functionalized monodisperse HILIC columns new HILIC column

Abstract

ABSTRACTIn this study, new hydroxyl-functionalized monodisperse polymeric hydrophilic interaction chromatography (HILIC) columns were developed using different derivatization agents. In addition, the influences of derivatization temperature of the best agent and polymer composition on the separation were investigated under HILIC conditions. Monodisperse–porous hydrophilic particles were synthesized by the seeded polymerization method using 3-chloro-2-hydroxypropile methacrylate (HPMA-CL) and ethylene glycol dimethacrylate (EGDMA) monomers. The chloropropyl terminal ends of the poly(HPMA-Cl-co-EGDMA) particles were derivatized with amine group of ethanolamine (EA), diethanolamine, and triethanolamine (TEA) at 80°C through nucleophilic reaction. The performance of synthesized particles was evaluated with the amount of ligand on the particle surfaces, column backpressure, and separation power under HILIC condition. TEA was found to be the best derivatization agent for the separation of toluene, acrylamide, thymine, adenine, and cytosine in respect to resolution factors (>1.5 for all analytes) and theoretical plate numbers (64.562 N/m for acylamide). Upon determination of the best ligand, then the effect of different derivatization temperatures and polymer composition on TEA performance was investigated. Of all the tested polymer compositions, the chromatographic performance of TEA-M-80 (the derivatization of TEA at 80°C together with M polymer composition) was found to be the best.