MACROPOROUS POLYMERIC STATIONARY-PHASE ROD AS CONTINUOUS SEPARATION MEDIUM FOR REVERSED-PHASE CHROMATOGRAPHY
MACROPOROUS POLYMERIC STATIONARY-PHASE ROD AS CONTINUOUS SEPARATION MEDIUM FOR REVERSED-PHASE CHROMATOGRAPHY
复制标题
DOI：
10.1021/ac00065a013
复制
发表时间：
1993-09-01
影响因子：
7.4
通讯作者：
FRECHET, JMJ
中科院分区：
文献类型：
--
作者：
WANG, QC;SVEC, F;FRECHET, JMJ
大孔聚（苯乙烯-二乙烯基苯）棒是通过在色谱柱范围内对含有单体、引发剂和致孔溶剂的混合物进行自由基聚合制备的，并首次用于蛋白质的极快速反相高效液相色谱法。通过压汞孔隙率测定法对连续棒的孔结构进行表征，发现存在大量直径约1μm至100nm以下的孔。尺寸排阻色谱法和扫描电子显微镜证实了不寻常的孔径分布。大孔的存在使得杆很容易被洗脱液渗透，因此，即使在高流速下，杆柱的背压也是适度的。聚合柱的效率几乎与流速无关。确定了几种蛋白质的显示容量因子与流动相组成的关系线的斜率，并开发了用于分离其混合物的梯度。正如分辨率数据所记录的，即使在 25 mL/min 的高流速下也能实现出色的分离。将柱长度增加三倍并没有提高蛋白质分离中的柱分辨率。
A macroporous poly(styrene-co-divinylbenzene) rod has been prepared by a free-radical polymerization of a mixture containing monomers, initiator, and porogenic solvent in the confines of a chromatographic column and used for the first time in the very fast reversed-phase HPLC of proteins. Characterization of the pore structure of the continuous rod by mercury intrusion porosimetry revealed a large volume of pores with a diameter of about 1 mum to pores below 100 nm. Size exclusion chromatography and scanning electron microscopy confirmed the unusual pore size distribution. The presence of large pores make the rod easily permeable to eluents, and therefore, the back pressure of the rod column is modest even at high flow rates. The efficiency of the polymerized column is almost independent of the flow rate. The slope of the line showing capacity factor vs composition of the mobile phase was determined for several proteins, and a gradient for the separation of their mixtures was developed. Excellent separation was achieved even at a high flow rate of 25 mL/min as documented by the resolution data. Tripling the length of the column did not improve the column resolution in protein separation.