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Application of accelerated solvent extraction coupled with high-performance counter-current chromatography to extraction and online isolation of chemical constituents from Hypericum perforatum L.
- Zhang, Yuchi, Liu, Chunming, Yu, Min, Zhang, Zhengkun, Qi, Yanjuan, Wang, Jing, Wu, Guimei, Li, Sainan, Yu, Jing, Hu, Yang
- Journal of chromatography 2011 v.1218 no.20 pp. 2827-2834
- Hypericum perforatum, chemical composition, chemical structure, chlorogenic acid, countercurrent chromatography, high performance liquid chromatography, hydrophilicity, hydrophobicity, hyperforin, nuclear magnetic resonance spectroscopy, raw materials, solvents
- Accelerated solvent extraction (ASE) coupled with high-performance counter-current chromatography (HPCCC) was successfully used for the extraction and online isolation of five chemical constituents from the plant Hypericum perforatum L. The upper phase of the solvent system of ethyl acetate–methanol–water (5:2:5, v:v:v) was used as both the ASE solvent and the HPCCC stationary phase. Two hydrophobic compounds including 28.4mg of hyperforin with a HPLC purity of 97.28% and 32.7mg of adhyperforin with a HPLC purity of 97.81% were isolated. The lower phase of ethyl acetate–methanol–n-butanol–water (5:2:2.5:12, v:v:v:v) was used as both the ASE solvent and CCC stationary phase. Three hydrophilic compounds of 12.7mg of 3,4,5-O-tricaffeoylquinic acid with a HPLC purity of 98.82%, 15.2mg of 1,3,5-O-tricaffeoylquinic acid with a HPLC purity of 99.46% and 42.5mg of 3-O-caffeoylquinic acid with a HPLC purity of 96.90%, were obtained in a one-step extraction–separation process with less than 3h from 10.02g of raw material of H. perforatum. The targeted compounds isolated, collected and purified by HPCCC were analyzed by high performance liquid chromatography (HPLC), the chemical structures of all five compounds above mentioned were identified by UV, MS and NMR.