以自行合成的8.0 μm大孔单分散亲水性交联聚甲基丙烯酸环氧丙酯微球为基质,采用化学改性方法合成了邻二醇配基新型高效疏水作用色谱填料.详细评价了该填料对蛋白质的分离性能、动力学吸附容量、质量回收率、硫酸铵起始浓度和流动相pH值对蛋白保留的影响.结果表明,这种高效疏水作用色谱填料的分离性能优良,分离了5种标准蛋白,对溶菌酶的活性回收率为96%±5%,吸附容量高达36.5 g/L.
Based on the 8.0 μm monodisperse poly (glycidyl methacrylate-co-ethylenedimethacrylate) beads (PGMA/EDMA) with macroperous as a media, a new hydrophobic interaction chromatographic (HIC) stationary phase for high performance liquid chromatography (HPLC) was synthesized by reaction of the hydroxyl groups of hydrolyzed macroperous PGMA/EDMA beads with epichlorohydrin followed by another hydrolysis of the newly introduced epexide groups. The stationary phase was evaluated with the properties of separability, dynamic loading capacity, mass recovery, effect of concentration of ammonium sulfate and pH on the retention of proteins in detail. It is found that it follows HIC retention mechanism. The measured bioactivity recovery for lysozyme was (96 ± 5 )%. The dynamic protein loading capacity of the synthesized HIC packings was 36.5 mg/g. Five proteins were completyly separated with 20 min linear gradient elution using the synthesized HIC stationary phase.