Bushey M M, Jorgenson J W
Department of Chemistry, University of North Carolina, Chapel Hill 27599-3290.
Anal Chem. 1990 Jan 15;62(2):161-7. doi: 10.1021/ac00201a015.
A comprehensive two-dimensional (2-D) liquid chromatographic separation system is presented. The system uses a microbore cation exchange column, operated under gradient conditions, as the first dimension separation. Effluent from this first column alternately fills one of two loops on a computer-controlled eight-port valve. A second pump then forces loop material onto a second column, a size exclusion column. UV detection is used, and the system is applied to the separation of protein standards and serum proteins. The 2-D system has a higher resolving power and peak capacity than either of the two columns used alone. The entire first column effluent is analyzed on the second column in virtually the same time it takes to complete the first column separation, without the use of stopped flow methods. The entire system is automated and operated under computer control. Three-dimensional (3-D) data representation provides a means of viewing peak profiles in either separation dimension and contour mapping of the 3-D data provides a more reliable means of peak identification from run to run than that provided by single-column elution times.
本文介绍了一种综合二维（2-D）液相色谱分离系统。该系统使用在梯度条件下运行的微径阳离子交换柱作为第一维分离。第一根色谱柱的流出物交替填充计算机控制的八通阀上的两个定量环之一。然后，第二个泵将定量环中的物质泵入第二根色谱柱，即尺寸排阻色谱柱。采用紫外检测，该系统应用于蛋白质标准品和血清蛋白的分离。二维系统比单独使用的两根色谱柱中的任何一根都具有更高的分离能力和峰容量。在完成第一根色谱柱分离所需的几乎相同时间内，无需使用停流方法，即可在第二根色谱柱上对第一根色谱柱的全部流出物进行分析。整个系统是自动化的，并在计算机控制下运行。三维（3-D）数据表示提供了一种查看任一分离维度上的峰形图的方法，并且与单柱洗脱时间相比，3-D数据的等高线图提供了一种在不同运行之间更可靠的峰识别方法。
