超高效液相串联质谱法测定大鼠血浆中胡麻属苷浓度

目的: 建立超高效液相串联质谱法快速测定大鼠体内胡麻属苷的浓度。方法: 用乙腈沉淀蛋白方法处理血浆,色谱柱为ACQUITY UPLC BEH C₁₈ 柱(50 mm×2.1 mm,1.7 μm),流动相为乙腈-0.1% 甲酸水,梯度洗脱,流速为0.4 mL·min^-1;用正离子多离子反应监测(MRM)扫描,内标为芦丁。结果: 血浆中胡麻属苷的线性范围为1~250 ng·mL^-1(r=0.998 6),最低定量限为0.2 ng·mL^-1;胡麻属苷的日内、日间RSD 均<10%。大鼠单次灌胃胡麻属苷后,胡麻属苷在大鼠体内的t_1/2 为(1.35±0.26)h,CL 为(8.62±1.25)L·h^-1·kg^-1,AUC_0-t 为(349.57±48.5)ng·h·mL^-1;大鼠舌下静注胡麻属苷后,胡麻属苷在大鼠体内的t_1/2 为(1.45±0.28)h,CL 为(2.38±0.57)L·h^-1·kg^-1,AUC_0-t 为(128.79±30.52)ng·h·mL^-1,得胡麻属苷的绝对生物利用度为26.6%。结论: 该方法经方法学验证,适于大鼠血浆中胡麻属苷浓度的测定及其药代动力学研究。

Objective: To develop an ultra performance liquid chromatography-tandem mass spectrometry method for the content determination of sesamoside in rat plasma. Methods: Plasmas were precipitated with 200μL acetonitrile. An ACQUITY UPLC BEH C₁₈(50 mm×2.1 mm,1.7 μm)column was used as the stationary phase. The mobile phase was consisted of acetonitrile and 0.1% formic acid water with gradient elution at a flow rate of 0.4 mL·min^-1. The analytes were detected with positive electrospray ionization in multiple reaction monitoring(MRM)mode and rutin was used as internal standard. Results: Excellent linear calibration curve of sesamoside was obtained in the concentration range of 1-250 ng·mL^-1(r=0.998 6). The lowesr limit of quantification was 0.2 ng·mL^-1. t_1/2 of sesamoside in rats was(1.35±0.26)h,CL of sesamoside in rats was (8.62±1.25)L·h^-1·kg^-1,AUC_0-t of sesamoside in rats was(349.57±48.5)ng·h·mL^-1 following intragastric administration of sesamoside;t_1/2 of sesamoside in rats was(1.45±0.28)h,CL of sesamoside in rats was (2.38±0.57)L·h^-1·kg^-1,AUC_0-t of sesamoside in rats was(128.79±30.52)ng·h·mL^-1 following sublingual intravenous administration of sesamoside,whose absolute bioavailabilityrate was 26.6%. The intra and inter day relative standard deviations of sesamoside were all below 10%. Conclusion: The established method is proved by methodology validation that it is suitable for pharmacokinetics study of sesamoside in rats.