基于超高效液相色谱-质谱联用法检测大鼠血浆中的钩吻绿碱和常绿钩吻碱及其药动学研究

Determination of gelsevirine and sempervirine in rat plasma based on ultra-performance liquid chromatography-tandem mass spectrometry and study on its pharmacokinetics

目的研究在口服5 mg/kg和舌下静脉注射0.1 mg/kg 2种不同给药方式下,钩吻绿碱和常绿钩吻碱在大鼠体内的药代动力学差异。方法色谱柱为UPLC BEH C_(18)(50 mm×2.1 mm,1.7μm),柱温设为40℃。以甲醇-水(含0.1%甲酸)为流动相,梯度洗脱,流速设为0.4 ml/min,洗脱时间为5 min。电喷雾(ESI)正离子模式检测,多反应监测(MRM)模式对钩吻绿碱m/z 353.1→322.1,常绿钩吻碱m/z 273.2→257.0和内标m/z 335.2→184.2进行定量分析。结果经口服给药后,钩吻绿碱的t_(1/2)为(13.8±13.7)h,AUC_((0-t))为(215.8±96.2)ng/(ml·h),CL_(z)为(21.2±10.0)L/(h·kg);舌下静脉注射给药后,钩吻绿碱的t_(1/2)为(11.7±13.9)h,AUC_((0-t))为(70.5±10.9)ng/(ml·h),CLz为(1.2±0.5)L/(h·kg)。经口服给药后,常绿钩吻碱的t_(1/2)为(5.5±4.8)h,AUC_((0-t))为(44.9±18.7)ng/(ml·h),CLz为(113.4±64.3)L/(h·kg);舌下静脉注射给药后,常绿钩吻碱的t_(1/2)为(7.6±4.1)h,AUC_((0-t))为(23.6±6.5)ng/(ml·h),CLz为(4.0±1.2)L/(h·kg)。结论建立UPLC-MS/MS技术测定大鼠血浆中钩吻绿碱和常绿钩吻碱的方法并成功应用于药动学研究,生物利用度低,分别为6.1%、3.8%。

Objective To study the pharmacokinetics of gelsevirine and sempervirine in rat plasma under two different administration ways(oral administration of 5 mg/kg and intravenous administration of 0.1 mg/kg).Methods The chromatographic column was UPLC BEH C_(18)(50 mm×2.1 mm,1.7μm),and the column temperature was set at 40℃.The mobile phase was methanol-water(containing 0.1%formic acid),the flow rate was 0.4 ml/min,and elution time was 5 min.Multiple reaction monitoring(MRM)in electrospray ionization(ESI)positive mode was used for quantitative analysis,m/z 353.1→322.1 for gelsevirine,m/z 273.2→257.0 for sempervirine and m/z 335.2→184.2 for internal standard.Results After oral administration,the t_(1/2)of gelsevirine was(13.8 viri)h,AUC_((0-t))was(215.8±96.2)ng/(ml·h),and CLzwas(21.2±10.0)L/(h·kg);after intravenous injection,the t_(1/2)of gelsevirine was(11.7±13.9)h,the AUC_((0-t))was(70.5±10.9)ng/(ml·h),and the CLzwas(1.2±0.5)L/(h·kg).After oral administration,the t_(1/2)of the sempervirine was(5.5±4.80)h,the AUC_((0-t))was(44.9±18.7)ng/(ml·h),and the CLzwas(113.4±64.3)L/(h·kg);after intravenous injection,the t_(1/2)of the sempervirine was(7.6±4.1)h,the AUC_((0-t))was(23.6±6.5)ng/(ml·h),and the CLzwas(4.0±1.2)L/(h·kg).Conclusion A UPLC-MS/MS method was established for the determination of gelsevirine and sempervirine in rat plasma and was successfully applied to pharmacokinetic studies.The bioavailability of gelsevirine and sempervirine was low,6.1%and 3.8%,respectively.