不同剂量薯蓣皂苷元的大鼠体内药动学研究

Study on pharmacokinetics of diosgenin at different doses in rats

目的:研究不同剂量薯蓣皂苷元的大鼠体内药物动力学特征,为阿尔茨海默病的临床前研究提供理论依据。方法:大鼠灌胃低(17.5 mg·kg^-1)、中(35 mg·kg^-1)、高剂量(70 mg·kg^-1)薯蓣皂苷元后,于不同时间眼眶取血,以延龄草苷作为内标,血浆样品经液-液萃取后采用LC-MS/MS进行测定。利用DAS 2.0计算药动学参数。色谱、质谱条件:采用Sun Fire C18(150 mm×2.1 mm,5μm)为色谱柱,以甲醇-乙腈^-10 mmol·L^-1乙酸铵(86∶11∶3)为流动相,流速为0.3 mL·min^-1,柱温40℃。采用电喷雾离子源(ESI源),以选择反应监测(MRM)方式并在正离子模式下进行检测,用于定量的离子对分别为m/z 415.5→271.5(薯蓣皂苷元)和m/z 577.5→271.5(内标延龄草苷)。结果:薯蓣皂苷元血药浓度在1~2000 ng·mL^-1内线性关系良好,方法回收率在81.7%~83.9%之间,定量下限(LLOQ)为1 ng·mL^-1,日内、日间精密度RSD小于10%。药动学结果表明不同剂量薯蓣皂苷元药-时曲线符合二室模型,主要药动学参数为AUC0-t分别为1872,3144,6625 ng·min·mL^-1;Cmax分别为136,276,470 ng·mL^-1;t1/2分别为5.73,5.31,6.42 h。结论:薯蓣皂苷元在17.5~70.0 mg·kg^-1范围内其药动学行为呈线性相关,无动力学行为差异。

OBJECTIVE To study the pharmacokinetics of diosgenin at different doses in rats and to provide theoretical basis for the pre-clinical study of Alzheimer’s disease.METHODS After oral administration of diosgenin at different doses,serial blood samples were obtained.Tillin was employed as an internal standard,and the plasma sample was treated by liquid-liquid extraction.Then the blood concentration of diosgenin was determined by LC-MS/MS and the pharmacokinetic parameters were calculated by DAS 2.0.HPLC-MS/MS CONDITION The mobile phase consisted of methanol-acetonitrile^-10 mmol·L^-1 aqueous ammonium acetate(86∶11∶3)which was pumped at 0.3 mL·min^-1.The analytical column(150 mm×2.1 mm,5μm)was packed with C18 material and the column temperature was set at 40℃.The ion pairs used for quantification were m/z 415.5→271.5(diosgenin)and m/z 577.5→271.5(internal standard tillin)using an electrospray ionization source(ESI source)in a selected reaction monitoring(MRM)manner and detection in positive ion mode.RESULTS The linearity of diosgenin in plasma was good from 1 to 2000 ng·mL^-1.The recovery of the method was between 81.7%-83.9%.The lower limit of quantitation(LLOQ)was 1 ng·mL^-1(LLOQ).The pharmacokinetic results showed that the concentration-time curves of different doses of diosgenin fitted the two-compartment model.The main pharmacokinetic parameters were as follows:AUC0-t was 1872,3144 and 6625 ng·min·mL^-1;Cmax was 136,276 and 470 ng·mL^-1;t1/2 was 5.73,5.31 and 6.42 h.CONCLUSION The pharmacokinetic behavior of diosgenin was linearly correlated within the range of 17.5-70 mg·kg^-1.