School of Light Industry and Chemical Engineering, Dalian Polytechnic University, Dalian, 116034, China; Laboratory of Bioseparation Technology, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, 20892, USA.
School of Light Industry and Chemical Engineering, Dalian Polytechnic University, Dalian, 116034, China.
J Chromatogr A. 2018 May 25;1551:69-74. doi: 10.1016/j.chroma.2018.03.052. Epub 2018 Mar 28.
The intermittently pressed tubing was introduced in type-I counter-current chromatographic system as the separation column to improve the separation performance in the present study. The separations were performed with two different solvent systems composed of 1-butanol-acetic acid-water (4:1:5, v/v) (BAW) and hexane-ethyl acetate-methanol-0.1 M HCl (1:1:1:1, v/v) (HEMW) using dipeptides and DNP-amino acids as test samples, respectively. The chromatographic performance was evaluated in terms of retention of the stationary phase (Sf), theoretical plate (N) and peak resolution (Rs). In general, the type-I planetary motion with the multilayer coil of non-modified standard tubing can yield the best separation at a low revolution speed of 200 rpm with lower flow rate. The present results with intermittently pressed tubing indicated that the performance was also optimal at the revolution speed of 200 rpm where the lower flow rate was more beneficial to retention of stationary phase and resolution. In the moderately hydrophobic two-phase solvent system composed of hexane-ethyl acetate-metanol-0.1 M hydrochloric acid (1:1:1:1, v/v), DNP-amino acids were separated with Rs at 1.67 and 1.47, respectively, with 12.66% of stationary phase retention at a flow rate of 0.25 ml/min. In the polar solvent system composed of 1-butanol-acetic acid-water (4:1:5, v/v), dipeptide samples were resolved with Rs at 2.18 and 18.75% of stationary phase retention at a flow rate of 0.25 ml/min. These results indicate that the present system substantially improves the separation efficiency of type-I counter-current chromatographic system.
本文将间歇加压管引入到 I 型逆流色谱系统中作为分离柱，以改善分离性能。分别以 1-丁醇-乙酸-水（4:1:5，v/v）（BAW）和正己烷-乙酸乙酯-甲醇-0.1 M HCl（1:1:1:1，v/v）（HEMW）两个不同的溶剂体系，以二肽和 DNP-氨基酸为测试样品进行分离。通过固定相（Sf）、理论塔板数（N）和峰分辨率（Rs）来评价色谱性能。通常，使用非改性标准管的多层线圈的 I 型行星运动可以在低转速 200 rpm 下产生最佳分离，且流量较低。使用间歇加压管的本研究结果表明，在转速 200 rpm 下，流量较低更有利于固定相保留和分辨率，性能也是最佳的。在由正己烷-乙酸乙酯-甲醇-0.1 M 盐酸（1:1:1:1，v/v）组成的中等疏水性两相溶剂体系中，DNP-氨基酸的分离 Rs 分别为 1.67 和 1.47，在 0.25 ml/min 的流速下，固定相保留率分别为 12.66%。在由 1-丁醇-乙酸-水（4:1:5，v/v）组成的极性溶剂体系中，二肽样品的分离 Rs 分别为 2.18 和 18.75%，在 0.25 ml/min 的流速下，固定相保留率分别为 18.75%。这些结果表明，本系统大大提高了 I 型逆流色谱系统的分离效率。
