UPLC-MS/MS法用于大鼠血浆中20(S)-原人参二醇药动学研究

Determination of 20(S)-Protopanaxdiol by UPLC-MS/MS and Its Application in Pharmacokinetics in Rat Plasma

目的:建立沉淀蛋白法结合超高效液相色谱-三重四级杆质谱法灵敏检测大鼠血浆中20(S)-原人参二醇的分析方法。方法:血浆样品用0.05%甲酸/乙腈沉淀蛋白处理后,采用Waters ACQUITY BEH C18(50 mm×2.1 mm,1.7μm)色谱柱进行分离,以乙腈-水(80∶20)为流动相进行等度洗脱,在电喷雾负离子模式下,采用多反应监测模式进行定性定量分析。结果:20(S)-原人参二醇在1~2500 ng/mL浓度范围内呈现良好线性(r≥0.998),定量限为0.5 ng/mL;方法批内与批间精密度均小于15%,准确度RE在±12.2%以内;方法提取回收率在84.8%~94.1%;大鼠单次灌胃20(S)-原人参二醇样品后,血浆中20(S)-PPD的C_(max)、t_(max)、t_(1/2)和AUC_(0-∞)分别为3520 ng/mL、2 h、10.65 h和21760 ng·mL^(-1)·h^(-1)。结论:本方法准确度、灵敏度高,样品前处理方法简单、快速,可用于血浆样品中20(S)-原人参二醇的药动学研究。

Objective:A promising detection approach,based on protein precipitation method coupled by ultra performance liquid chromatography-triple quadrupole mass spectrometry,was established for determination of 20(S)-protopanaxdiol in rat plasma and using in pharmacokinetics study.Methods:20(S)-protopanaxdiol was effectively extracted by acetonitrile and separated at the Waters ACQUITY BEH C18 column(50 mm×2.1 mm,1.7μm)using acetonitrile-water(80∶20)as the mobile phase.Under the electrospray negative ion mode,the multi-reaction monitoring mode was used for qualitative and quantitative analysis for 20(S)-protopanaxdiol.Results:The results indicated that 20(S)-protopanaxdiol in plasma had a good linear relationship in a wide range of 1-2500 ng/mL and a lower detect limit of 0.5 ng/mL.The intra-and inter-batch precision were less than 15%,the accuracy RE was within±12.2%and the recoveries were 84.8%-94.1%for spiked plasma.Finally,the method was successfully used in pharmacokinetics of 20(S)-protopanaxdiol in rat plasma.The results showed C_(max) in plasma was 3520 ng/mL,t_(max) was 2 h,t_(1/2) was 10.65 h and AUC_(0-∞)was 21760 ng·mL^(-1)·h^(-1).Conclusion:This method is accurate,sensitive,simple and fast and can use for determination of 20(S)-protopanaxdiol and application in its pharmacokinetics.