On-line two-dimensional countercurrent chromatography×high performance liquid chromatography system with a novel fragmentary dilution and turbulent mixing interface for preparation of coumarins from Cnidium monnieri.
On-line two-dimensional countercurrent chromatography×high performance liquid chromatography system with a novel fragmentary dilution and turbulent mixing interface for preparation of coumarins from Cnidium monnieri.
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DOI：
10.1016/j.chroma.2015.06.034
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发表时间：
2015-08
期刊：
影响因子：
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通讯作者：
Dong Wang;Long-Jiang Chen;Jing-Lan Liu;Xin-yuan Wang;Yunlong Wu;Meijuan Fang;Zhen Wu;Y. Qiu
中科院分区：
文献类型：
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作者：
Dong Wang;Long-Jiang Chen;Jing-Lan Liu;Xin-yuan Wang;Yunlong Wu;Meijuan Fang;Zhen Wu;Y. Qiu
建立了一种新的在线二维逆流色谱×高效液相色谱（2D CCC × HPLC）系统，用于蛇床子中香豆素类化合物的一步制备分离。选择了具有合适的Kd值和较高的固定相保留值的正庚烷/丙酮/水（31：50：19，v/v）两相溶剂体系分离目标化合物。为了解决CCC和RP-HPLC之间的溶剂不相容性问题，成功开发了一种新型的分段稀释和湍流混合（FD-TM）接口。详细地，来自第一维CCC柱的洗脱液被来自补充泵的稀释水分成级分以在两个切换样品回路中形成“样品-稀释”条带。随后，应用长细管通过管内湍流将CCC洗脱液与水混合，以减少溶剂效应。将每个CCC级分交替捕获在两个保持柱上用于进一步制备型HPLC分离。这种国家设计的FD-TM策略有效地降低了柱后压力，并允许在CCC后末端具有更高的水稀释比，从而提高了样品回收率和稳健的2D CCC × HPLC分离系统。结果，在单次2D分离运行（6.5h）中，从0.5g的C. monnieri，总产率为1.3、2.0、0.5、0.5、0.8、1.5、8.2和15.0%，HPLC纯度分别为90.1、91.1、94.7、99.1、99.2、98.2、97.9和91.9%。基于FD-TM界面的改进型二维CCC × HPLC系统，有望在其他复杂植物源天然产物的多组分同时分离纯化中得到广泛应用。
This study describes a novel on-line two-dimensional countercurrent chromatography × high performance liquid chromatography (2D CCC × HPLC) system for one-step preparative isolation of coumarins from the fruits ofCnidium monnieri. An optimal biphasic solvent system composed ofn-heptane/acetone/water (31:50:19, v/v) with suitableKdvalues and a higher retention of the stationary phase was chosen to separate target compounds. In order to address the solvent incompatibility problem between CCC and RP-HPLC, a novel fragmentary dilution and turbulent mixing (FD-TM) interface was successfully developed. In detail, the eluent from the first dimensional CCC column was divided into fractions to form ‘sample-dilution’ stripes in the two switching sample loops, by the dilution water from the makeup pump. Following this, a long, thin tube was applied to mix the CCC eluent with water by in-tube turbulence, to reduce the solvent effect. Each CCC fraction was alternately trapped on the two holding columns for further preparative HPLC separation. This nationally designed FD-TM strategy effectively reduced post-column pressure and allowed a higher water dilution ratio at the post end of CCC, leading to improved sample recovery and a robust 2D CCC × HPLC isolation system. As a result, in a single 2D separation run (6.5 h), eight target compounds (1–8) were isolated from 0.5 g crude extract ofC. monnieri, in overall yields of 1.3, 2.0, 0.5, 0.5, 0.8, 1.5, 8.2, and 15.0%, with HPLC purity of 90.1, 91.1, 94.7, 99.1, 99.2, 98.2, 97.9, and 91.9%, respectively. We anticipate that this improved 2D CCC × HPLC system, based on the novel FD-TM interface, has broad application for simultaneous isolation and purification of multiple components from other complex plant-derived natural products.