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Novel Design for Centrifugal Counter-Current Chromatography: VI. Ellipsoid Column
Journal of Liquid Chromatography & Related Technologies ( IF 1.0 ) Pub Date : 2014-10-02 , DOI: 10.1080/10826076.2014.883533
Dongyu Gu 1 , Yi Yang 2 , Xuelei Xin 3 , Haji Akber Aisa 3 , Yoichiro Ito 4
Affiliation  

A novel ellipsoid column was designed for centrifugal counter-current chromatography. Performance of the ellipsoid column with a capacity of 3.4 mL was examined with three different solvent systems composed of 1-butanol–acetic acid–water (4:1:5, v/v/v) (BAW), hexane–ethyl acetate–methanol-0.1 M HCl (1:1:1:1, v/v/v/v) (HEMH), and 12.5% (w/w) PEG1000 and 12.5% (w/w) dibasic potassium phosphate in water (PEG-DPP) each with suitable test samples. In dipeptide separation with BAW system, both stationary phase retention (Sf) and peak resolution (Rs) of the ellipsoid column were much higher at 0° column angle (column axis parallel to the centrifugal force) than at 90° column angle (column axis perpendicular to the centrifugal force), where elution with the lower phase at a low flow rate produced the best separation yielding Rs at 2.02 with 27.8% Sf at a flow rate of 0.07 mL/min. In the DNP-amino acid separation with HEMW system, the best results were obtained at a flow rate of 0.05 ml/min with 31.6% Sf yielding high Rs values at 2.16 between DNP-DL-glu and DNP-β-ala peaks and 1.81 between DNP-β-ala and DNP-L-ala peaks. In protein separation with PEG-DPP system, lysozyme and myolobin were resolved at Rs of 1.08 at a flow rate of 0.03 mL/min with 38.9% Sf. Most of those Rs values exceed those obtained from the figure-8 column under similar experimental conditions previously reported.

中文翻译:

离心逆流色谱的新设计:VI。椭圆柱

一种新型椭圆柱被设计用于离心逆流色谱。使用由 1-丁醇-乙酸-水 (4:1:5, v/v/v) (BAW)、己烷-乙酸乙酯-组成的三种不同溶剂系统检查容量为 3.4 mL 的椭球柱的性能。甲醇-0.1 M HCl (1:1:1:1, v/v/v/v) (HEMH), 和 12.5% (w/w) PEG1000 和 12.5% (w/w) 磷酸氢二钾水溶液 (PEG -DPP) 每个都有合适的测试样本。在使用 BAW 系统的二肽分离中,椭球柱的固定相保留 (Sf) 和峰分辨率 (Rs) 在 0° 柱角(柱轴平行于离心力)下比在 90° 柱角(柱轴垂直于离心力),在低流速下用较低相洗脱产生最佳分离,Rs 为 2.02,Rs 为 27。8% Sf,流速为 0.07 mL/min。在使用 HEMW 系统的 DNP-氨基酸分离中,在 0.05 ml/min 的流速下获得了最佳结果,Sf 为 31.6%,在 DNP-DL-glu 和 DNP-β-ala 峰之间的 Rs 值为 2.16 和 1.81 DNP-β-ala 和 DNP-L-ala 峰之间。在使用 PEG-DPP 系统进行蛋白质分离时,溶菌酶和肌红蛋白在 0.03 mL/min 的流速和 38.9% 的 Sf 下以 1.08 的 Rs 分离。大多数 Rs 值超过了在之前报道的类似实验条件下从图 8 列中获得的值。在使用 PEG-DPP 系统进行蛋白质分离时,溶菌酶和肌红蛋白在 0.03 mL/min 的流速下以 1.08 的 Rs 分离,Sf 为 38.9%。大多数 Rs 值超过了在之前报道的类似实验条件下从图 8 列中获得的值。在使用 PEG-DPP 系统进行蛋白质分离时,溶菌酶和肌红蛋白在 0.03 mL/min 的流速下以 1.08 的 Rs 分离,Sf 为 38.9%。大多数 Rs 值超过了在之前报道的类似实验条件下从图 8 列中获得的值。
更新日期:2014-10-02
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