Development of a rapid and convenient method to separate eight ginsenosides from Panax ginseng by high-speed counter-current chromatography coupled with evaporative light scattering detection

다국어 초록 (Multilingual Abstract)

Ginsenosides are well-known major components isolated from the radix of Panax ginseng C. A. Meyer, known as Korean ginseng, having diverse biological activities. They have recently gained much attention for their biomedical applications. In the present work, a fast and simple method for the separation and purification of 8 ginsenosides from Panax ginseng by counter current chromatography coupled with evaporative light scattering detector (CCC-ELSD) was successfully established. The crude samples for CCC separation were first purified from ginseng extract using a macroporous resin. The extract was loaded onto a Diaion-HP20 column and fractionated by methanol and water gradient elution. The ginsenosides-triols and diols fractions were subsequently eluted with 65% & 80% methanol and water gradient elution, respectively. Furthermore, these two fractions were separated by CCC-ELSD. The two phase solvent system used for separation was composed of chloroform-methanol-water-isopropanol at a volume ratio of 4:3:2:1. Each fraction obtained was collected and dried, which yielded eight ginsenosides namely, Rg1, Re, Rf, Rh1, Rb1, Rc, Rb2 & Rd. The purity of these ginsenosides was more than 97% assessed by HPLC-ELSD system, and their structures were characterized by electrospray-ionization mass spectrometry (ESI-MS), 1H NMR and 13C NMR spectroscopy. This is the first report regarding the CCC separation of ginsenosides Rh1, Rb2 & Rc from Panax ginseng.