title: 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.
abstract: Ginsenosides exhibit diverse biological activities and are major well-known components isolated from the radix of Panax ginseng C.A. Meyer. In the present work, a rapid and facile method for the separation and purification of eight ginsenosides from P. ginseng by high-speed counter-current chromatography coupled with evaporative light scattering detector (HSCCC-ELSD) was successfully developed. The crude samples for HSCCC 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-protopanaxadiol (PPD) and protopanaxatriol (PPT) fractions were subsequently eluted with 65 and 80% methanol and water gradient elution, respectively. Furthermore, these two fractions were separated by HSCCC-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, yielding the following eight ginsenosides: Rg(1), Re, Rf, Rh(1), Rb(1), Rc Rb(2) and Rd. The purity of these ginsenosides was greater than 97% as assessed by HPLC-ELSD, and their structures were characterized by electrospray-ionization mass spectrometry (ESI-MS) and nuclear magnetic resonance spectroscopy. This is the first report regarding the separation of the ginsenosides Rh(1), Rb(2) and Rc from P. ginseng by HSCCC.
authors.affiliation: College of Pharmacy, Natural Products Research Institute, Seoul National University, Seoul, Korea.
authors.forename: Omer
authors.initials: O
authors.lastname: Shehzad
authors.forename: In Jin
authors.initials: IJ
authors.lastname: Ha
authors.forename: Youmie
authors.initials: Y
authors.lastname: Park
authors.forename: Young Wan
authors.initials: YW
authors.lastname: Ha
authors.forename: Yeong Shik
authors.initials: YS
authors.lastname: Kim
mesh_terms: D003377:Countercurrent Distribution / Q000295:instrumentation / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000737:chemistry / Q000302:isolation & purification*; D036145:Ginsenosides / Q000737:chemistry / Q000302:isolation & purification*; D015394:Molecular Structure; D005894:Panax / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D004365:Drugs, Chinese Herbal; D036145:Ginsenosides