Spiral disk assembly for HSCCC: Column design and basic studies on chromatographic resolution and stationary phase retention
Spiral disk assembly for HSCCC: Column design and basic studies on chromatographic resolution and stationary phase retention
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DOI：
10.1081/jlc-120021255
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发表时间：
2003-01-01
影响因子：
1.3
通讯作者：
Sullivan, JV
中科院分区：
文献类型：
--
作者：
Ito, Y;Yang, FQ;Sullivan, JV
为提高极性溶剂体系固定相的保留率，设计了一套四块螺旋通道的J型高速逆流色谱分离盘。测试了四个不同的螺旋盘：两个具有不同深度的单个螺旋通道，另外两个具有串联连接的四个螺旋通道以提供更大的螺旋节距。使用三种不同的两相溶剂系统，包括1-丁醇/乙酸/水，在色谱分离度和/或固定相保留方面测试每个盘的性能（4：1：5，v/v/v）用于二肽分离; 12.5%（w/w）聚乙二醇（PEG）1000-12.5%（w/w）磷酸氢二钾用于蛋白分离;和4%（w/w）PEG 8000 -5%（w/w）葡聚糖T500的10 mM磷酸氢二钠溶液，用于测定固定相保留。结果表明，即使在相对高的移动的相流速下（其中螺距起重要作用），对于所有溶剂系统，螺旋柱仍保留令人满意的固定相量。使用10 mL/min流速的移动的相和具有较大螺距的4-螺旋盘，用丁醇溶剂系统分离二肽是可能的。在使用PEG磷酸盐系统的蛋白质分离中，单螺旋盘在1 mL/min下使用尾对头洗脱模式的上相产生最佳分离，而4螺旋盘显示出更高的固定相保留。PEG-右旋糖酐体系的保留在4-螺旋盘中得到改善，在0.5 mL/min的流速下超过60%。讨论了影响螺旋盘分离柱性能的各种参数。
A set of four separation disks equipped with spiral channel(s) was designed for our type-J high speed countercurrent chromatography (J-HSCCC) centrifuge to improve retention of the stationary phase of polar solvent systems. Four different spiral disks were tested: two had a single spiral channel with different depths and the other two had four spiral channels connected in series to provide a greater spiral pitch. Performance of each disk was tested in terms of chromatographic resolution and/or stationary phase retention using three different two-phase solvent systems, including 1-butanol/acetic acid/water (4:1:5, v/v/v) for dipeptide separation; 12.5% (w/w) polyethylene glycol (PEG) 1000-12.5% (w/w) dibasic potassium phosphate for protein separation; and 4% (w/w) PEG8000-5% (w/w) dextran T500 in 10 mM dibasic sodium phosphate for determination of stationary phase retention. The results show that the spiral column retains a satisfactory amount of stationary phase for all solvent systems, even at a relatively high mobile phase flow rate where the spiral pitch plays a significant role. Separation of dipeptides with the butanol solvent system was possible using 10 mL/min flow rate of the mobile phase and 4-spiral-disks with their greater spiral pitch. In protein separations with the PEG phosphate system, the single-spiral disks yielded the best separation using the upper phase in tail-to-head elution mode at I mL/min, while the 4-spiral disks show higher retention of the stationary phase. The retention of the PEG-dextran system is improved in the 4-spiral disks, which exceed 60% at a How rate of 0.5 mL/min. Various parameters, which affect the performance of the spiral disk separation column are discussed.