Cao Xueli, Hu Guanghui, Huo Liangsheng, Zhu Xiaoping, Li Ting, Powell Jimmie, Ito Yoichiro
Beijing Technology and Business University, Beijing Key Lab of Plant Resource Research, Beijing 100037, China.
J Chromatogr A. 2008 Apr 25;1188(2):164-70. doi: 10.1016/j.chroma.2008.02.073. Epub 2008 Mar 4.
A spiral disk assembly composed of five single-channel units was designed for high-speed counter-current chromatography (HSCCC). The retention of different solvent systems ranging from moderately polar to polar organic-aqueous systems to aqueous two-phase systems (ATPs) was investigated under different elution modes. The results indicated that the spiral disk assembly can produce excellent retention of stationary phase for moderately polar organic-aqueous solvent systems, such as chloroform-methanol-water (4:3:2) and hexane-ethyl acetate-methanol-water (1:1:1:1) by pumping lower mobile phase from head (H) to tail (T), and upper mobile phase from tail (T) to head (H) even at a high flow-rate (8 mL/min, Sf>70%), regardless of whether the inlet is at the inner or outer terminal of the channel. This makes it possible for fast analysis of some small molecular compounds. This has been proved in the separation of mixtures of three flavones, including isorhamnetin, kaempferol, and quercetin. The spiral disk assembly can also provide satisfactory retention for polar to ATPS such as 1-butanol-acetic acid-water (4:1:5) (<3 mL/min, Sf>70%), 12.5% poly(ethylene glycol) (PEG) 1000-12.5% K2HPO4-75% water (< or =1 mL/min, Sf>70%) and 4% PEG 8000-5% Dextran T500-91% water (< or =0.5 mL/min, Sf>50%) by pumping lower mobile phase from inner terminal (I) to outer terminal (O), and upper mobile phase from outer terminal (O) to inner terminal (I) at a low flow-rate, while this is not possible with the multilayer coil column. Acceptable resolutions were achieved when it was used for the separation of peptides such as Leu-Tyr and Val-Tyr, and proteins including cytochrome c and myoglobin, lysozyme and myoglobin, and fresh chicken egg-white proteins.
设计了一种由五个单通道单元组成的螺旋盘组件用于高速逆流色谱（HSCCC）。研究了在不同洗脱模式下，从中等极性到极性有机-水体系再到双水相体系（ATPs）等不同溶剂体系的保留情况。结果表明，对于中等极性有机-水溶剂体系，如氯仿-甲醇-水（4:3:2）和己烷-乙酸乙酯-甲醇-水（1:1:1:1），通过从头部（H）向尾部（T）泵送较低流速的流动相，以及从尾部（T）向头部（H）泵送较高流速的流动相，即使在高流速（8 mL/min，Sf>70%）下，螺旋盘组件也能实现对固定相的出色保留，无论入口位于通道的内端还是外端。这使得快速分析某些小分子化合物成为可能。这在分离包括异鼠李素、山柰酚和槲皮素在内的三种黄酮混合物中得到了证明。对于极性至ATPS体系，如1-丁醇-乙酸-水（4:1:5）（<3 mL/min，Sf>70%）、12.5%聚乙二醇（PEG）1000 - 12.5%磷酸氢二钾-75%水（≤1 mL/min，Sf>70%）和4% PEG 8000 - 5%葡聚糖T500 - 91%水（≤0.5 mL/min，Sf>50%），通过在低流速下从内端（I）向外端（O）泵送较低流速的流动相，以及从外端（O）向内端（I）泵送较高流速的流动相，螺旋盘组件也能提供令人满意的保留，而多层螺旋柱则无法做到这一点。当用于分离如亮氨酸-酪氨酸和缬氨酸-酪氨酸等肽类以及包括细胞色素c和肌红蛋白、溶菌酶和肌红蛋白以及新鲜鸡蛋白蛋白等蛋白质时，获得了可接受的分离度。
