Lignans of Schisandra chinensis: isolation and analysis in plant cell cultures

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Title in English Future trends in Phytochemistry, A young scientists symposium, Book of abstracts
Authors

BŘEZINOVÁ Lenka VLAŠÍNOVÁ Helena HAVEL Ladislav HUMPA Otakar CHVÁTALOVÁ Kateřina SLANINA Jiří

Year of publication 2007
Type Article in Proceedings
Conference Book of abstracts
MU Faculty or unit

Faculty of Medicine

Citation
Field Analytic chemistry
Keywords Schisandra; lignans; HPLC; plant culture;
Description The fruit of Schisandra chinensis, a woody liana, has been used for centuries in traditional Chinese medicine. The fruit is prescribed for the treatment of hepatitis in China. Active principles are lignans with unusual structures derived from dibenzo[a,c]cyclooctadiene. These lignans have been shown to possess a broad range of biological effects. To obtain pure standards of lignans, 349 g of Schisandra chinensis were extracted with petroleum ether. The fraction rich in lignans was obtained by extraction of petroleum ether fraction with methanol. Methanol extract was fractionated on silica gel column and subsequently by high performance liquid chromatography on reversed phase columns 250 x 25 and 250 x 8 mm (Tessek SGX C18, Prague) with methanol and water as a mobile phase. Seven lignans, schizandrin, tigloylgomisin H, gomisin A, deoxyschizandrin, gamma-schizandrin, gomisin N and wuweizisu C, were isolated. Their structures were identified by spectral methods (1D and 2D NMR, UV, MS) and confirmed with those published previously. High performance liquid chromatography coupled with ultraviolet detection was used for the determination of lignans in the samples of cell cultures of Schisandra chinensis. Methanol was found to be the most efficient solvent for the extraction of lignans from freeze-dried samples. Methanolic extracts and media samples were cleaned by solid-phase extraction with Strata C18-E (Phenomenex) cartridges. After the loading of the samples, the cartridges were washed with 40 % aqueous methanol (v/v) to remove interfering compounds. The lignans were eluted from the cartridges with methanol and examined by HPLC. The chromatographic separation was carried out on Chromolith Performance RP-18e monolith column (100 x 4.6 mm, Merck) using isocratic mobile phase of acetonitrile and water in the ratio 50:50 (v/v). The base line separation of lignans was achieved within a short time (20 minutes) owing to use of the monolithic column, which enables the high flow rate of mobile phase (2 ml/min).
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