Ito Yoichiro, Clary Robert
Laboratory of Bioseparation Technology, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892. USA.
Machine Instrumentation Design and Fabrication, National Institutes of Health, Bethesda, MD 20892, USA.
Separations. 2016 Dec;3(4). doi: 10.3390/separations3040031. Epub 2016 Oct 14.
High-speed countercurrent chromatography with a spiral tube assembly can retain a satisfactory amount of stationary phase of polymer phase systems used for protein separation. In order to improve the partition efficiency a simple tool to modify the tubing shapes was fabricated, and the following four different tubing modifications were made: intermittently pressed at 10 mm width, flat, flat-wave, and flat-twist. Partition efficiencies of the separation column made from these modified tubing were examined in protein separation with an aqueous-aqueous polymer phase system at flow rates of 1-2 ml/min under 800 rpm. The results indicated that the column with all modified tubing improved the partition efficiency at a flow rate of 1 ml/min, but at a higher flow rate of 2 ml/min the columns made of flattened tubing showed lowered partition efficiency apparently due to the loss of the retained stationary phase. Among all the modified columns, the column with intermittently pressed tubing gave the best peak resolution. It may be concluded that the intermittently pressed and flat-twist improve the partition efficiency in a semi-preparative separation while other modified tubing of flat and flat-wave configurations may be used for analytical separations with a low flow rate.
采用螺旋管组件的高速逆流色谱法能够保留用于蛋白质分离的聚合物相体系中足量的固定相。为提高分配效率，制作了一种用于改变管形状的简易工具，并进行了以下四种不同的管形状改变：以10毫米宽度间歇挤压、扁平状、扁平波浪状和平扭状。在800转/分钟的转速下，以1 - 2毫升/分钟的流速，用双水相聚合物相体系对由这些改变形状后的管制成的分离柱在蛋白质分离中的分配效率进行了考察。结果表明，所有改变形状后的管制成的柱在1毫升/分钟的流速下提高了分配效率，但在2毫升/分钟的较高流速下，由扁平管制成的柱由于保留的固定相损失而明显显示出分配效率降低。在所有改变形状后的柱中，间歇挤压管制成的柱给出了最佳的峰分辨率。可以得出结论，间歇挤压管和平扭状管在半制备分离中提高了分配效率，而其他扁平状和扁平波浪状的改变形状后的管可用于低流速的分析分离。
