液-质联用研究五味子醇甲、五味子甲素和五味子乙素在大鼠体内的组织分布

目的: 建立HPLC-MS同时测定大鼠组织中五味子醇甲、五味子甲素和五味子乙素的分析方法,并应用于3个木脂素成分在大鼠体内的组织分布研究。 方法: 以睾丸酮为内标,组织样品经乙腈沉淀蛋白后进行HPLC-MS分析,采用依利特ODS-C₁₈色谱柱(250 mm×4.6 mm,5 μm),乙腈(A)-0.05%甲酸水溶液(B)为流动相,梯度洗脱;正离子选择性检测(SIM)模式,选择的检测离子质荷比(m/z)为五味子醇甲433.2、五味子甲素401.0、五味子乙素417.0和睾丸酮289.0。 结果: 在大鼠不同组织中,3个木脂素成分在测定的线性范围内线性关系良好(r≥0.9951);方法的精密度和稳定性RSD均小于15%;回收率为81.11%~99.49%,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。3个木脂素成分在大鼠体内分布比较广泛,主要分布在肝和肺组织中,在脾中的分布浓度最低。 结论: 建立的方法简便易行、准确可靠,应用于大鼠体内木脂素成分的组织分布研究,获得了满意的结果。

Objective: To develop an HPLC-MS method for simultaneous determination of schisandrin,deoxyschisandrin and schisandrin B in rat tissues,and apply the method to tissue distribution study of three studied lignans. Methods: The tissue samples were precipitated with acetonitrile and analyzed by HPLC-MS with acetonitrile-0.05% formic acid solution as a gradient elution mobile phase.Testosterone was used as an internal standard.The mass spectrometer was operated in selected reaction monitoring(SIM)mode with positive electrospray ionization,the transition m/z 433.2,m/z 401.0,m/z 417.0 and m/z 289.0 was used for schisandrin,deoxyschisandrin,schisandrin B and testosterone,respectively. Results: Excellent linearity(r≥0.9951)was observed in all three 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 81.11%-99.49% with RSD less than 15%.Schisandrin,deoxyschisandrin and schisandrin B were widely distributed in rat tissues.Majority was found in liver and lung,and lowest in spleen. Conclusion: The established method is simple and accurate.Satisfactory results are obtained applying this method to the tissue distribution study of schisandrin,deoxyschisandrin and schisandrin B.