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Preparative Isolation of six Anti‐Tumour Biflavonoids from Selaginella Doederleinii Hieron by High‐Speed Counter‐Current Chromatography
Author(s) -
Li Shaoguang,
Zhao Meifeng,
Li Yuxiang,
Sui Yuxia,
Yao Hong,
Huang Liying,
Lin Xinhua
Publication year - 2013
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/pca.2478
Subject(s) - biflavonoids , chemistry , amentoflavone , selaginella , petroleum ether , dichloromethane , chromatography , ethyl acetate , countercurrent chromatography , ether , chloroform , solvent , high performance liquid chromatography , organic chemistry , extraction (chemistry) , stereochemistry , botany , biology
Biflavonoids are the primary constituents of Selaginella doederleinii Hieron, to which different bioactivities have been attributed, including anti‐cancer, anti‐inflammatory, anti‐oxidant, anti‐fungal and anti‐virus activity. However, effective methods for separation of these compounds are not currently available. Objective To develop a high performance and bioassay‐guided method for preparative isolation of biflavonoids from S . doederleini via high‐speed counter‐current chromatography (HSCCC). Methods The anti‐proliferation effects of four fractions (70% ethanol, petroleum ether, dichloromethane and acetic ether extracts) of S . doederleinii on five human cancer cells were monitored. The dichloromethane and acetic ether extracts showed good cytotoxicities to the studied cancer cell lines, guiding the subsequent separation. Two solvent systems composed of n ‐hexane:ethyl acetate:methanol:water (1:2:1.5:1.5, v/v) and n ‐hexane:ethyl acetate:methanol:water (3:2:3:2, v/v) were developed for separation of the active fractions, respectively. Identification of the biflavonoids was performed by EI‐MS n , 1 H‐ and 13 C‐NMR.' Results Under the optimised conditions, 12.6 mg amentoflavone (91.4%), 6.6 mg robustaflavone (90.4%), 7.5 mg 2'', 3''‐dihydro‐3', 3'''‐biapigenin (98.2%) and 7.3 mg 3', 3'''‐binaringenin (90.3%) from acetic ether extract (500 mg) and 6.3 mg heveaflavone (93.5%) and 5.3 mg 7, 4', 7'', 4'''‐tetra‐ O ‐methyl‐amentoflavone (94.5%) from dichloromethane extract (200 mg) were obtained, respectively. The anti‐proliferation effects of the six biflavonoids on the five human cancer cells were further verified. Conclusion The study provides methodological references for simultaneously preparative isolation of several bioactive biflavones from the herbal family of Selaginella . It is the first report discovering 2'', 3''‐dihydro‐3', 3'''‐biapigenin and 3', 3'''‐binaringenin from this herb and describing their cytotoxicities to human cancer cell lines. Copyright © 2013 John Wiley & Sons, Ltd.