Moore A W, Jorgenson J W
Department of Chemistry, University of North Carolina-Chapel Hill 27599, USA.
Anal Chem. 1995 Oct 1;67(19):3448-55. doi: 10.1021/ac00115a013.
Coupled-column two dimensional (2D) separation systems offer potentially high peak capacity and are amenable to automation. Reversed phase liquid chromatography (RPLC) and capillary zone electrophoresis (CZE) are complementary techniques well suited for use in a 2D system. Optically gated CZE is a means of performing rapid CZE analyses. In a 2D system, these rapid CZE analyses enable more frequent sampling of the RPLC separation and thus more freedom in control of the RPLC analysis conditions. Complete 2D separations can be done in the time usually required to do the RPLC analysis alone. With the present system, a complete 2D analysis can be done in under 10 min. Because the peak capacity of a 2D method is inherently high, some of the available peak capacity can be exchanged for speed of analysis. Acceleration of the RPLC elution gradient will decrease analysis time, but RPLC resolution will suffer. However, because it is a 2D system, some of the resolution lost in the RPLC can be regained in the second dimension CZE analysis. Here, the RPLC gradient was done over only 2 min, but CZE analyses done every 2.5 s restore some of the resolution lost in the rapid RPLC gradient.
联用柱二维（2D）分离系统具有潜在的高柱效，并且适合自动化操作。反相液相色谱（RPLC）和毛细管区带电泳（CZE）是非常适合用于二维系统的互补技术。光控CZE是一种进行快速CZE分析的方法。在二维系统中，这些快速CZE分析能够更频繁地对RPLC分离进行采样，从而在RPLC分析条件的控制上拥有更大的自由度。完整的二维分离可以在通常仅进行RPLC分析所需的时间内完成。使用当前系统，完整的二维分析可以在10分钟内完成。由于二维方法的柱效本质上很高，一些可用的柱效可以用来换取分析速度。加快RPLC洗脱梯度会减少分析时间，但RPLC分辨率会受到影响。然而，因为这是一个二维系统，在RPLC中损失的一些分辨率可以在第二维CZE分析中恢复。在这里，RPLC梯度仅在2分钟内完成，但每2.5秒进行一次的CZE分析恢复了在快速RPLC梯度中损失的一些分辨率。
