液-质联用法同时测定L-02肝细胞中五味子甲素和五味子乙素含量

Simultaneous determination of deoxyschisandrin and schisandrin B in L-02 cell by HPLC-MS

目的:建立HPLC-MS同时测定L-02肝细胞中五味子甲素和五味子乙素的分析方法,并应用于研究2个木脂素成分在L-02肝细胞的摄取过程。方法:以睾丸酮为内标,细胞样品经乙腈沉淀蛋白后进行HPLC-MS分析,采用依利特ODS-C_(18)色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.05%甲酸水溶液(45∶55)为流动相,流速1.0 mL·min^(-1);正离子选择性检测(SIM)模式,选择的检测离子质荷比(m/z)为五味子甲素401.0、五味子乙素417.0和睾丸酮289.0。结果:在L-02肝细胞中,2个木脂素成分在测定的线性范围内线性关系良好(r≥0.996 6);方法的精密度和稳定性RSD均小于15%;回收率为94.99%~100.8%,RSD小于15%,符合化学药物非临床药代动力学研究技术指导原则。2个木脂素成分均能够被L-02肝细胞所摄取,并呈现先升高后下降的趋势。结论:本文方法应用于研究2种木脂素成分在L-02肝细胞的摄取过程,获得了满意的结果。

Objective： To develop an HPLC-MS method for schisandrin B in L-02 cell. The method was applied to study the simultaneous determination of deoxyschisandrin and absorption of two lignans in L-02 cell. Methods： The cell samples were precipitated with acetonitrile and analyzed by HPLC-MS with acetonitrile-0.05% formic acid solution （ 45 ： 55 ）. Testosterone was used as an internal standard. The mass spectrometer was operated in selected reaction monitoring （SIM）mode with positive electrospray ionization, the transitions m/z 401.0, m/z 417.0 and m/z 289.0 were used for deoxyschisandrin, schisandrin B and testosterone, respectively. Results： Excellent linearity （ r ≥ 0,996 6 ） was observed in two studied lignans in their linear range. The RSD of precision and stability of the developed method was less than 15%. The recovery of the method was 94.99%-100.8% with RSD less than 15%. Deoxyschisandrin and schisandrin B were absorbed in L-02 cell. The concentration was increased first and then decreased with time. Conclusion： Satisfactory results were obtained by applying this method to the cell absorption study of deoxyschisandrin and schisandrin B.