Brezinová Lenka, Vlasínová Helena, Havel Ladislav, Humpa Otakar, Slanina Jirí
Department of Biochemistry, Masaryk University, Brno, Czech Republic.
Biomed Chromatogr. 2010 Sep;24(9):954-60. doi: 10.1002/bmc.1391.
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 x 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, gamma-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, gamma-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.
建立并验证了一种用于测定五味子植物细胞培养物中六种木脂素的简单快速方法。木脂素用甲醇从植物样品中提取，提取物用Strata C18 - E（Phenomenex）柱进行固相萃取有效净化。色谱分离在Chromolith Performance RP - 18e整体柱（100×4.6 mm，默克）上进行，使用乙腈和水体积比为50:50的等度流动相。在220 nm处监测洗脱液。五味子醇甲、戈米辛A、脱氧五味子醇甲、γ-五味子醇甲、戈米辛N和五味子素C在相对较短的时间内（20分钟）实现了基线分离，这得益于流动相相对较高的流速（2 mL/min）。五味子醇甲和戈米辛A的定量下限为0.1 mg/L，脱氧五味子醇甲、γ-五味子醇甲和戈米辛N为0.3 mg/L，五味子素C为1 mg/L。对含有六种木脂素的加标样品进行分析，在所有情况下绝对回收率在93%至101%之间。该验证方法成功应用于五味子胚性植物细胞培养物中木脂素的测定。
