Urban Jiří, Jandera Pavel, Langmaier Pavel
Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Pardubice, Czech Republic.
J Sep Sci. 2011 Aug;34(16-17):2054-62. doi: 10.1002/jssc.201100174. Epub 2011 May 19.
The polarity of (poly)methacrylate monolithic capillary columns was varied by using alkylmethacrylate monomers with butyl, cyclohexyl, 2-ethylhexyl, lauryl, and stearyl functional groups in the polymerization mixture. The hydrodynamic properties, as well as the retention characteristics in RP-LC of small molecules (alkylbenzenes) and of proteins under gradient elution conditions were studied. The RP selectivity depends on the type of alkyl chain in methacrylate monomer; however, there was no direct correlation between the size of the monomer molecule and methylene or aromatic selectivity of the monlithic column. The lowest selectivity was found for column based on lauryl methacrylate monomer. On the other hand, butyl methacrylate column shows high phenyl selectivity and the column with stearyl methacrylate possesses the highest methylene selectivity for small molecules. The retention increases with longer alkyl chain in methacrylate monomer, especially for high molar mass proteins on all prepared columns and showed gradient elution behavior of proteins in agreement with the linear solvent strength gradient model. The poly(laurylmethacrylate) column showed lowest hydrophobicity but best efficiency for proteins of all columns tested.
通过在聚合混合物中使用具有丁基、环己基、2-乙基己基、月桂基和硬脂基官能团的甲基丙烯酸烷基酯单体，改变了（聚）甲基丙烯酸酯整体毛细管柱的极性。研究了小分子（烷基苯）和蛋白质在梯度洗脱条件下的流体动力学性质以及在反相液相色谱中的保留特性。反相选择性取决于甲基丙烯酸酯单体中烷基链的类型；然而，单体分子的大小与整体柱的亚甲基或芳香族选择性之间没有直接相关性。基于月桂基甲基丙烯酸酯单体的柱的选择性最低。另一方面，丁基甲基丙烯酸酯柱对苯基具有高选择性，而硬脂基甲基丙烯酸酯柱对小分子具有最高的亚甲基选择性。随着甲基丙烯酸酯单体中烷基链变长，保留增加，特别是对于所有制备的柱上的高摩尔质量蛋白质，并且蛋白质表现出与线性溶剂强度梯度模型一致的梯度洗脱行为。在所有测试的柱中，聚（月桂基甲基丙烯酸酯）柱对蛋白质的疏水性最低但效率最佳。
