Capillary Electrochromatographic Separation of Proteins with Covalently Bonded Ionic Liquid Column
Date Issued
2008
Date
2008
Author(s)
Wang, Hui-Ling
Abstract
To prepare ionic liquid columns, the capillary was first silanzed with (3-chloropropyl)trimethoxysilane (CPTMS). Imidazole was then subjected to be bonded covalently to the CPTMS. Finally, bonded imidazole in the column was linked with 1-bromooctane to form a positively charged diakylimidazolium layer for the generation of reversed electroosmotic flow (EOF) over a pH range of 4 to 8.5. Using phosphate as background electrolyte (BEG), the strong ion-pair interaction between diakylimidazolium and phosphate caused the bleeding of stationary phase. We selected five proteins, such as ovalbumin (OVA), apo-transferrin (apoTf), bovine serum albumin (BSA), rat serum albumin (RSA) and conalbumin (CON) as model compounds, whose molecular weights and pI values are alike to each other. Both acetate and formate buffer have been systematically studiced as mobile phase. Formate buffer gave better performance than acetate buffer. With running buffer of formate (pH 3.5, 40 mM) and effective length of 85 cm, five proteins showed four absorption peaks within 40 min with RSD ≤ 4.59% (n = 3). In this study, the retention behavior of imidazole coated column was also compared with those of the ionic liquid column. The best is the C8 substituent of imidazolium ionic liquid. High resolving power of the ionic liquid column seems to be predominantly from hydrophobic force, hydrogen bonding, π-π electron and electrostatic interactions.
Subjects
capillary electrochromatographic separation
ionic liquid
protein
Type
thesis
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