RAPID COMPREHENSIVE 2-DIMENSIONAL SEPARATIONS OF PEPTIDES VIA RPLC OPTICALLY GATED CAPILLARY ZONE ELECTROPHORESIS
RAPID COMPREHENSIVE 2-DIMENSIONAL SEPARATIONS OF PEPTIDES VIA RPLC OPTICALLY GATED CAPILLARY ZONE ELECTROPHORESIS
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DOI：
10.1021/ac00115a013
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发表时间：
1995-10-01
影响因子：
7.4
通讯作者：
JORGENSON, JW
中科院分区：
文献类型：
--
作者：
MOORE, AW;JORGENSON, JW
偶联柱二维（2D）分离系统提供潜在的高峰容量并且易于自动化。反相液相色谱（RPLC）和毛细管区带电泳（CZE）是互补的技术，适用于二维系统。光学门控CZE是进行快速CZE分析的一种手段。在2D系统中，这些快速CZE分析能够更频繁地对RPLC分离进行采样，从而更自由地控制RPLC分析条件。完全的2D分离可以在通常单独进行RPLC分析所需的时间内完成。使用本系统，可以在10分钟内完成完整的2D分析。由于2D方法的峰值容量固有地高，因此可以将一些可用的峰值容量交换为分析速度。加速RPLC洗脱梯度将减少分析时间，但RPLC分离度将受到影响。然而，因为它是一个2D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。这里，RPLC梯度仅在2分钟内完成，但每2.5秒进行一次CZE分析，恢复了快速RPLC梯度中损失的一些分辨率。
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 wed 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.