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Validated method for bioactive lignans in Schisandra chinensis in vitro cultures using a solid phase extraction and a monolithic column application
Authors:Lenka Březinová  Helena Vlašínová  Ladislav Havel  Otakar Humpa  Jiří Slanina
Institution:1. Department of Biochemistry, Faculty of Medicine, Masaryk University, Kamenice 5, Building A16, CZ‐62500 Brno, Czech Republic;2. Department of Plant Biology, Faculty of Agronomy, Mendel University of Agriculture and Forestry Brno, Zemědělská 1, CZ‐61300 Brno, Czech Republic;3. National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Kamenice 5, Building A4, CZ‐62500 Brno, Czech Republic
Abstract:A simple and rapid method for determination of six lignans found in plant cell cultures of Schisandra chinensis was developed and validated. The lignans were extracted from plant samples with methanol and the extracts were effectively cleaned by solid‐phase extraction using Strata C18‐E (Phenomenex) cartridges. Chromatographic separation was carried out on a Chromolith Performance RP‐18e monolithic column (100 × 4.6 mm, Merck) using an isocratic mobile phase of acetonitrile and water in a 50:50 (v/v) ratio. The eluent was monitored at 220 nm. The baseline separation of schizandrin, gomisin A, deoxyschizandrin, γ‐schizandrin, gomisin N and wuweizisu C was achieved in a relatively short time period (20 min), which was made possible by the relatively high flow rate of the mobile phase (2 mL/min). The lower limit of quantitation was 0.1 mg/L for schizandrin and gomisin A, 0.3 mg/L for deoxyschizandrin, γ‐schizandrin, and gomisin N and 1 mg/L for wuweizisu C. The analysis of spiked samples containing six lignans provided absolute recoveries between 93 and 101% in all cases. The validated method was successfully applied to the determination of lignans in embryogenic plant cell cultures of Schisandra chinensis. Copyright © 2010 John Wiley & Sons, Ltd.
Keywords:Schisandra chinensis  HPLC  monolithic column  dibenzocyclooctadiene lignans  plant cell culture
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