Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussel, Belgium.
Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussel, Belgium.
Anal Chim Acta. 2022 Jul 25;1218:339962. doi: 10.1016/j.aca.2022.339962. Epub 2022 May 21.
Separation performance in chromatography has been extensively studied since the dawn of the technique. Although the basic principles of band broadening and the resulting separation performance in isocratic elution are in general well known and understood, this is much less the case for gradient separations. In this tutorial, first the basic principles, concepts and parameters that determine separation performance, peak width and variance and analysis time in isocratic separations are reviewed. This is subsequently used to discuss the parameters that affect peak width in gradient elution, together with the concepts of plate count and plate height in this elution mode. In addition, the effect of peak compression in gradient elution is elaborated. Finally, the effect of extra-column dispersion on separation performance in gradient elution is discussed, and an overview of how these contributions can be experimentally evaluated is given.
自该技术出现以来，色谱分离性能已得到广泛研究。尽管在等度洗脱中带展宽的基本原理和由此产生的分离性能通常众所周知且已被充分理解，但在梯度分离中情况则要差得多。在本教程中，首先回顾了决定等度分离中分离性能、峰宽和方差以及分析时间的基本原理、概念和参数。随后，将使用这些原理来讨论影响梯度洗脱中峰宽的参数，以及在这种洗脱模式下的塔板数和塔板高度的概念。此外，还详细阐述了梯度洗脱中峰压缩的影响。最后，讨论了柱外扩散对梯度洗脱分离性能的影响，并概述了如何通过实验评估这些贡献。
