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Separation of triterpenoid saponins from the root of Bupleurum falcatum by counter current chromatography: The relationship between the partition coefficients and solvent system composition
Author(s) -
Lee Kyoung Jin,
Xu MeiYing,
Shehzad Omer,
Seo Eun Kyoung,
Kim Yeong Shik
Publication year - 2014
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201400871
Subject(s) - chemistry , chromatography , partition coefficient , bupleurum , elution , countercurrent chromatography , saponin , methanol , solvent , chloroform , column chromatography , organic chemistry , medicine , alternative medicine , pathology , traditional chinese medicine
A separation method using counter current chromatography coupled with an evaporative light‐scattering detection system was developed to purify five triterpenoid saponins from the roots of Bupleurum falcatum . The methanol extract was loaded onto a Diaion® HP20 column and fractionated by a methanol and water gradient elution. The saikosaponin‐enriched fraction was obtained by elution with 100% methanol. The two‐phase solvent systems used for separation were composed of chloroform/methanol/isopropanol/water at a volume ratio of 60:60:1:60 and 6:6:1:6. The relationship between the isopropanol ratio of each phase and the partition coefficients of the target compounds was investigated by calculating partition coefficient by high‐performance liquid chromatography and measuring the accurate composition of each phase by 1 H NMR spectroscopy. Each fraction obtained was collected and dried, which yielded the following five saikosaponins from 700 mg of injected sample: saikosaponin B 1 (8.7 mg), saikosaponin A (86 mg), saikosaponin B 3 (17 mg), saikosaponin B 2 (41 mg), and saikosaponin C (33 mg). Saikosaponin A showed the most potent cytotoxicity against human cancer cells (gastric cancer, AGS cells; breast cancer, MCF‐7 cells; and hepatoma, HepG2 cells) after 24 h. The IC 50 values for the above three cell types were 34.6, 33.3, and 23.4 μmol/L, respectively.