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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 establish an HPLC-MS method for the simultaneous determination of Schisandrin A and Schisandrin B in L-02 hepatocytes and to investigate the uptake of two lignans in L-02 hepatocytes. METHODS: Testosterone was used as an internal standard. The samples were precipitated by acetonitrile and analyzed by HPLC-MS. The elution was performed on a column of ODS-C 18 (250 mm × 4.6 mm, 5 μm) with acetonitrile-0.05% formic acid 45:55) was used as the mobile phase at a flow rate of 1.0 mL · min -1. In the SIM mode, the mass-to-charge ratio (m / z) of the test mass was 401.0 for Schisandrin, 417.0 and testosterone 289.0. Results: In L-02 hepatocytes, the linear relationship between the two lignans was linear (r≥0.996 6). The RSD of the method was less than 15% and the recovery was 94.99% ~ 100.8%, RSD less than 15%, in line with non-clinical pharmacokinetic study of technical guidelines. Both lignan components were uptake by L-02 hepatocytes and showed a trend of first increasing and then decreasing. Conclusion: The method proposed in this paper was applied to study the uptake of two lignan components in L-02 hepatocytes with satisfactory results.