UPLC-MS/MS法同时测定大鼠血浆中赤芍提取物三种单萜类成分及其药代动力学研究

Simultaneous Determination of Main Monoterpenoids in Radix Paeoniae Rubra Extract by UPLC-MS/MS Method and Analysis of Their Pharmacokinetics in Plasma of Rats

目的：研究赤芍提取物中3种有效成分芍药苷、芍药内酯苷和氧化芍药苷在大鼠体内药动学特点。方法：色谱采用Agilent Eclipse Plus C18色谱柱（4.6 mm×100 mm,3.5μm）,体积流量0.5 m L/min,流动相采用0.05%甲酸乙腈-0.05%甲酸水,梯度洗脱,质谱采用电喷雾电离源,扫描方式为多反应离子监测。大鼠一次性经皮注射赤芍提取物后,采用UPLC-MS/MS法测定大鼠血浆、胆汁、尿液、粪便中芍药苷、芍药内酯苷和氧化芍药苷的含量,用药动学软件DAS 3.0分析3种成分的药动学特点。结果：3种成分的峰面积和质量浓度之间呈良好线性关系（r〉0.99）,提取回收率在89.57%~111.21%之间,精密度、准确度、基质效应和稳定性均良好。结论：芍药苷和芍药内酯苷在大鼠体内具有吸收快,消除快的特点,氧化芍药苷半衰期稍长。这3种成分的主要排泄方式为粪便〉胆汁〉尿液。

Objective： To develop anultra performance liquid chromatography tandem mass sepctrometry（ UPLC-MS/MS）method for simultaneously determining the contents of paeoniflorin,albiflorin,Oxypaeoniflorin and benzoylpaeoniflorin in rat plasma and their pharmacokinetics. Methods： Four components were simultameously determined by UPLC with Agilent Eclipse Plus C18（ 3. 5 μm,4. 6 mm × 100 mm）. The mobile phase,consisting of acetonitrile and 0. 05% aqueous formic acid,was programmed as a linear gradient. The flow rate was 0. 5 m L/min. The components were ionized in the electrospray ionization ion source of the mass spectrometer and detected in the multiple reaction model. We determined the level of the four components in plasma,bile,urine and feces in rats by UPLC-MS/MS and analyzed the pharmacokinetic features of the four components by the pharmacokinetic software DAS3. 0. Results： The relationship between the concentration and the peak areas of the three components was all linear（ r 0. 99）. The precision,accuracy,extract recoveries,matrix effects and stability of the analytes met the requirements. Conclusion： The experiment indicates the four components distribute and eliminate rapidly in rats. The main way of excretion is confirmed by feces.