Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, China; Graduate University of the Chinese Academy of Sciences, Beijing 100049, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Nov 1;938:75-9. doi: 10.1016/j.jchromb.2013.08.012. Epub 2013 Sep 1.
High-speed counter-current chromatography (HSCCC) was successfully applied for the first time to isolate and purify four cis-trans isomers of coumaroylspermidine analogs from Safflower. HSCCC separation was achieved with a two-phase solvent system composed of chloroform-methanol-water (1:1:1, v/v/v) with the upper phase as the mobile phase. In a single run, a total of 1.3mg of N(1), N(5), N(10)-(E)-tri-p-coumaroylspermidine (EEE), 4.4mg of N(1)(E)-N(5)-(Z)-N(10)-(E)-tri-p-coumaroylspermidine (EZE), 7.2mg of N(1)(Z)-N(5)-(Z)-N(10)-(E)-tri-p-coumaroylspermidine (ZZE), and 11.5mg of N(1),N(5),N(10)-(Z)-tri-p-coumaroylspermidine (ZZZ) were obtained from 100mg of crude sample. High Performance Liquid Chromatography (HPLC) analysis showed that the purities of these four components are 95.5%, 98.1%, 97.5% and 96.2%, respectively. The chemical structures were identified by ESI-MS, (1)H NMR and (13)C NMR.
高速逆流色谱（HSCCC）首次成功应用于从红花中分离和纯化 4 种顺反式香豆酰基亚精胺类似物的异构体。HSCCC 分离是在氯仿-甲醇-水（1:1:1，v/v/v）两相溶剂系统中进行的，上相为流动相。在单次运行中，从 100mg 粗样品中总共得到 1.3mg 的 N(1)、N(5)、N(10)-(E)-三对香豆酰基亚精胺（EEE）、4.4mg 的 N(1)(E)-N(5)-(Z)-N(10)-(E)-三对香豆酰基亚精胺（EZE）、7.2mg 的 N(1)(Z)-N(5)-(Z)-N(10)-(E)-三对香豆酰基亚精胺（ZZE）和 11.5mg 的 N(1)、N(5)、N(10)-(Z)-三对香豆酰基亚精胺（ZZZ）。高效液相色谱（HPLC）分析表明，这四个成分的纯度分别为 95.5%、98.1%、97.5%和 96.2%。通过 ESI-MS、（1）H NMR 和（13）C NMR 确定了它们的化学结构。
