School of Pharmaceutical Sciences, University of Geneva, Geneva, Switzerland.
J Sep Sci. 2010 Mar;33(6-7):752-64. doi: 10.1002/jssc.200900749.
In recent years, numerous works have been reported on hydrophilic interaction chromatography (HILIC) columns, but there is a growing demand for fast separations with improved resolution. Such analyses can only be obtained with the new generation of columns and instrumentation. In the present study, HILIC columns packed with porous sub-2 microm particles or with superficially porous particles of 2.7 microm were evaluated. A set of 15 compounds with diverse physicochemical properties was selected. The aim of this study was to compare the chromatographic behavior of three selected columns. Concerning kinetic performance, the fused-core column offers a backpressure two times lower than the sub-2-microm particle columns. However, these superficially porous particles also generated a 30% lower efficiency. The complex mechanism of HILIC was also investigated on bare silica columns, confirming that the mechanism is a mixture of partitioning, adsorption and ion exchange in different proportions according to the nature of the analytes, the composition of the mobile phase and the characteristics of the analytical support. Finally, a rapid separation of three drugs and their corresponding metabolites was carried out. The analysis time was reduced to less than 4 min while maintaining acceptable backpressure and sufficient resolution.
近年来，已有大量关于亲水作用色谱（HILIC）柱的报道，但人们对快速分离和提高分辨率的需求日益增长。只有使用新一代的柱和仪器才能实现这种分析。本研究评估了填充有亚 2 微米多孔颗粒或 2.7 微米表面多孔颗粒的 HILIC 柱。选择了一组具有不同物理化学性质的 15 种化合物。本研究的目的是比较三种选定柱的色谱行为。关于动力学性能，核壳柱的背压比亚 2 微米颗粒柱低两倍。然而，这些表面多孔颗粒的效率也降低了 30%。在裸硅胶柱上还研究了复杂的 HILIC 机制，证实该机制是根据分析物的性质、流动相的组成和分析支持物的特性，以不同比例分配、吸附和离子交换的混合物。最后，对三种药物及其相应代谢物进行了快速分离。分析时间减少到不到 4 分钟，同时保持可接受的背压和足够的分辨率。
