Chen Xin, Tolley H Dennis, Lee Milton L
J Sep Sci. 2011 Aug;34(16-17):2063-71.
A stable poly(2-carboxyethyl acrylate-co-poly(ethylene glycol) diacrylate) monolith was synthesized inside a 75-μm id capillary by direct in situ photo-initiated polymerization in a binary porogenic solvent consisting of methanol and ethyl ether. The resulting monolith was evaluated for weak cation-exchange capillary liquid chromatography of peptides and proteins. A high dynamic binding capacity of 72.7 mg lysozyme per cm3 column volume was measured. Fast mass transfer was demonstrated by steep breakthrough curves. The resulting monolith exhibited negligible hydrophobicity, leading to good separation of peptides and proteins. Peak capacities of 11 for peptides with a 10-min salt gradient and 39 for proteins with a 20-min salt gradient were measured. An efficiency of 37,000 plates/m for proteins was obtained under isocratic conditions. The effects of functional group concentration, porogenic solvent composition, mobile phase pH, salt gradient rate, and hydrophobicity on the retention of analytes were investigated. Good run-to-run relative standard deviation (RSD) <1.93% and column-to-column RSD <4.63% were achieved.
通过在由甲醇和乙醚组成的二元致孔溶剂中直接原位光引发聚合，在内径75μm的毛细管内合成了一种稳定的聚（丙烯酸2 - 羧乙酯 - 共 - 聚乙二醇二丙烯酸酯）整体柱。对所得整体柱进行了用于肽和蛋白质的弱阳离子交换毛细管液相色谱分析评估。测得每立方厘米柱体积的溶菌酶高动态结合容量为72.7毫克。陡峭的穿透曲线证明了快速传质。所得整体柱的疏水性可忽略不计，从而实现了肽和蛋白质的良好分离。对于具有10分钟盐梯度的肽，测得的峰容量为11；对于具有20分钟盐梯度的蛋白质，测得的峰容量为39。在等度条件下，蛋白质的柱效为37,000理论塔板数/米。研究了官能团浓度、致孔溶剂组成、流动相pH值、盐梯度速率和疏水性对分析物保留的影响。实现了良好的批间相对标准偏差（RSD）<1.93%和柱间RSD<4.63%。
