Preparative separation and purification of four cis-trans isomers of coumaroylspermidine analogs from safflower by high-speed counter-current chromatography

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 1.3 mg of N-1, N-5, N-10-(E)-tri-p-coumaroylspermidine (EEE), 4.4 mg of N-1(E)-N-5-(Z)-N-10-(E)-tri-p-coumaroylspermidine (EZE), 7.2 mg of N-1(Z)-N-5-(z)-N-10-(E)-tri-p-coumaroylspermidine (ZZE), and 11.5 mg of N-1,N-5,N-10-(Z)-tri-p-coumaroylspermidine (ZZZ) were obtained from 100 mg 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, H-1 NMR and C-13 NMR. (c) 2013 Elsevier B.V. All rights reserved.