Russo Rossana, Guillarme Davy, T-T Nguyen Dao, Bicchi Carlo, Rudaz Serge, Veuthey Jean-Luc
Laboratory of Phytoanalysis, Dipartimento di Scienza e Tecnologia del Farmaco, Turin University, Via P. Giuria 9-10125 Turin, Italy.
J Chromatogr Sci. 2008 Mar;46(3):199-208. doi: 10.1093/chromsci/46.3.199.
During the last few years, there has been a great interest in the development of fast liquid chromatography. Among the reported approaches, the reduction of the particle size to attain sub-2microm diameter represents a good solution for achieving both increased separation power and faster analysis time. This paper demonstrates the chromatographic performance of such supports using plate-height curves and reveals the possibility for obtaining ultra-fast or highly efficient separations, according to the column geometry and system pressure limitations. The stability of these columns is initially evaluated using a system suitability experiment. The chromatographic performance remains stable in terms of retention, efficiency, and pressure for more than 1700 injections with pressure conditions ranging from 200 to 800 bar. Several fast and ultra-fast pharmaceutical applications are reported. In isocratic mode, a 5- to 10-fold reduction in analysis time is obtained with limited influence on efficiency and resolution. The run time is further reduced by 30-fold with the shorter available columns (i.e., 30 mm length). In gradient mode, the separation of a complex mixture containing an active pharmaceutical compound and related impurities is significantly improved with column length equal to 100 mm, to increase peak capacity and resolution.
在过去几年中，人们对快速液相色谱的发展产生了浓厚兴趣。在已报道的方法中，减小颗粒尺寸以获得直径小于2微米的颗粒是实现更高分离能力和更快分析时间的良好解决方案。本文使用板高曲线展示了此类载体的色谱性能，并揭示了根据柱几何形状和系统压力限制获得超快速或高效分离的可能性。这些色谱柱的稳定性最初通过系统适用性实验进行评估。在200至800巴的压力条件下，经过超过1700次进样，色谱性能在保留时间、柱效和压力方面保持稳定。本文还报道了一些快速和超快速的药物分析应用。在等度模式下，分析时间可减少5至10倍，而对柱效和分离度的影响有限。使用较短的可用色谱柱（即30毫米长度）时，运行时间可进一步减少30倍。在梯度模式下，对于含有活性药物成分和相关杂质的复杂混合物，使用100毫米长的色谱柱可显著改善分离效果，从而提高峰容量和分离度。
