University of Hohenheim, Institute of Food Chemistry, Department of Food Chemistry (170b), D-70593 Stuttgart, Germany.
University of Hohenheim, Institute of Food Chemistry, Department of Food Chemistry (170b), D-70593 Stuttgart, Germany.
J Chromatogr A. 2018 Jun 29;1556:88-96. doi: 10.1016/j.chroma.2018.04.069. Epub 2018 May 1.
Countercurrent chromatography (CCC) is an all liquid based separation technique typically used for the isolation and purification of natural compounds. The simplicity of the method makes it easy to scale up CCC separations from analytical to preparative and even industrial scale. However, scale-up of CCC separations requires two different instruments with varying coil dimensions. Here we developed two variants of the CCC multiple injection mode as an alternative to increase the throughput and enhance productivity of a CCC separation when using only one instrument. The concept is based on the parallel injection of samples at different points in the CCC column system and the simultaneous separation using one pump only. The wiring of the CCC setup was modified by the insertion of a 6-port selection valve, multiple T-pieces and sample loops. Furthermore, the introduction of storage sample loops enabled the CCC system to be used with repeated injection cycles. Setup and advantages of both multiple injection modes were shown by the isolation of the furan fatty acid 11-(3,4-dimethyl-5-pentylfuran-2-yl)-undecanoic acid (11D5-EE) from an ethyl ester oil rich in 4,7,10,13,16,19-docosahexaenoic acid (DHA-EE). 11D5-EE was enriched in one step from 1.9% to 99% purity. The solvent consumption per isolated amount of analyte could be reduced by ∼40% compared to increased throughput CCC and by ∼5% in the repeated multiple injection mode which also facilitated the isolation of the major compound (DHA-EE) in the sample.
逆流色谱（CCC）是一种基于全液体的分离技术，通常用于天然化合物的分离和纯化。该方法简单，易于从分析规模放大到制备规模，甚至工业规模。然而，CCC 分离的放大需要两种不同的仪器，其线圈尺寸也不同。在这里，我们开发了两种 CCC 多次进样模式的变体，作为在仅使用一种仪器时增加 CCC 分离通量和提高生产力的替代方法。该概念基于在 CCC 柱系统的不同点同时平行进样，并仅使用一台泵进行同时分离。通过插入 6 通选择阀、多个 T 型件和样品环来修改 CCC 装置的布线。此外，引入存储样品环使 CCC 系统能够用于重复进样循环。通过从富含 4,7,10,13,16,19-二十二碳六烯酸乙酯（DHA-EE）的乙酯油中分离出呋喃脂肪酸 11-（3,4-二甲基-5-戊基呋喃-2-基）-十一烷酸（11D5-EE）来展示两种多次进样模式的设置和优点。11D5-EE 一步从 1.9%纯度提高到 99%纯度。与增加的 CCC 通量相比，每个分析物的分离量的溶剂消耗可减少约 40%，在重复多次进样模式下也可减少约 5%，这也有利于样品中主要化合物（DHA-EE）的分离。
