高效液相-质谱联用法(LC-MS/MS)测定延龄草苷血药浓度及其药物动力学研究

Determination of trillin in mouse plasma by LC-MS/MS method and pharmacokinetic study

目的:进行延龄草苷在小鼠体内的药物动力学研究。方法:采用甲醇沉淀蛋白进行血浆样品预处理,选用人参皂苷 Rh2为内标,以 LC-MS/MS 法测定小鼠血浆中延龄草苷的含量。色谱柱为 MetaChem ODS-3(50mm×2.0mm,5μm),流动相为乙腈-水-甲酸(80:20:0.1),流速为0.2mL·min^(-1)。电喷雾离子化(ESI)方式,采用多反应监测,检测离子为正离子,分别选择 m/z 577.6→253.6amu 和 m/z 621.5→603.7amu 作为延龄草苷和内标物人参皂苷 Rh2的检测离子对。结果:延龄草苷在2.0μg·mL^(-1)～200μg·mL^(-1)范围内线性关系良好(r=0.9964)。方法的准确度范围为-3.0%～1.4%,日内精密度 RSD 范围为1.1%～3.3%,日间精密度 RSD 范围为3.2%～5.1%。小鼠尾静脉注射延龄草苷(7.6mg·kg^(-1))后,药-时数据符合二室模型,主要药物动力学参数:t_(1/2)为5.56h、K_e 为0.12h^(-1)、C_(max)为130.1μg·mL^(-1)、V_d 为4.6mL、Cl 为0.049mL·h·kg^(-1)。结论:建立的含量测定方法快速、准确,适用于延龄草苷在小鼠体内的药物动力学研究。

Objective：To investigate pharmacokinetics of trillin in mouse. Method：Trillin was determined after the methanol - mediated plasma protein precipitation. Ginsenoside Rh2 was used as internal standard. A LC - MS/MS method was established for the determination of trillin in mouse plasma. Metachem ODS -3 column （50mm×2.0min,5μm）was used as analytical column. The mobile phase consisted of acetonitrile water formic acid（ 80： 20： 0. 1, v/v/v）, which was pumped at 0. 2mL·min^-1. The analytes were detected using electro - spray ionization （ESI） interface with positive ionization. Ions were monitored in the multiple reaction -monitoring（MRM） modes, the fragmentation transitions were m/z 577.6→253.6amu for trillin and m/z 621.5→03.7amu for ginsenoside Rh2. Result：The method was linear（ r =0. 9964）within the range of 2. 0 -200 μg·mL^-1. The accuracy of method was range from-3.0% to 1.4% ,the intra- day and inter-day relative standard deviation（ RSD）were in the range of 1.1% -3.3% and 3.2% -5.1%. This disposition was conformed a two - compartment model after intravenous administrations（at a dose of 7.6 mg·kg^-1） to mice. The main pharmacokinetic parameters were： t1/2 ,5.56 h; Ke, 0. 12 h^-1; Cmax130. 1μg·mL^-1 ;Va,4. 6 mL; Cl,0. 049 mL·h·kg^-1. Conclusion：This method is simple and rapid, accurate, specific and precise. It is applicable for pharmacokinetic study of trillin in mouse.