UPLC-MS/MS method for the simultaneous quantification of pravastatin, fexofenadine, rosuvastatin, and methotrexate in a hepatic uptake model and its application to the possible drug-drug interaction study of triptolide
UPLC-MS/MS method for the simultaneous quantification of pravastatin, fexofenadine, rosuvastatin, and methotrexate in a hepatic uptake model and its application to the possible drug-drug interaction study of triptolide
复制标题
UPLC-MS/MS 方法同时定量肝摄取模型中的普伐他汀、非索非那定、瑞舒伐他汀和甲氨蝶呤及其在雷公藤甲素可能的药物相互作用研究中的应用
DOI：
10.1002/bmc.5093
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发表时间：
2021
影响因子：
1.8
通讯作者：
Huang Xin
中科院分区：
文献类型：
--
作者：
Wu Wei;Cheng Rui;Jiang Zhenzhou;Zhang Luyong;Huang Xin
开发了一种快速、特异的UPLC-MS/MS方法，总运行时间为3.5 min，用于测定大鼠原代肝细胞中的普伐他汀、非索非那定、瑞舒伐他汀和甲氨蝶呤。用70%乙腈（含30%H2O）沉淀蛋白质后，在梯度条件下，以0.03%乙酸（v/v）和甲醇（v/v）组成的移动的流动相，流速为0.50 mL/min，分离4种分析物。方法的线性、回收率、基质效应、准确度、精密度和稳定性得到了良好的验证。我们基于这四种化合物在新鲜悬浮的肝细胞中评价了药物-药物相互作用。在4°C下，普伐他汀、非索非那定、瑞舒伐他汀和甲氨蝶呤的肝摄取显著低于37°C，并且肝细胞随着底物浓度和培养时间的增加而饱和，表明大鼠原代肝细胞模型成功建立。雷公藤内酯醇对这四种化合物的肝摄取有明显的抑制作用。总之，该方法成功用于定量测定普伐他汀、非索非那定、瑞舒伐他汀和甲氨蝶呤，并用于验证Oatp 1、Oatp 2、Oatp 4和Oat 2转运蛋白研究的大鼠原代肝细胞模型。然后，我们应用该模型探讨雷公藤甲素对这四种转运蛋白的影响。
A rapid and specific UPLC–MS/MS method with a total run time of 3.5 min was developed for the determination of pravastatin, fexofenadine, rosuvastatin, and methotrexate in rat primary hepatocytes. After protein precipitation with 70% acetonitrile (containing 30% H2O), these four analytes were separated under gradient conditions with a mobile phase consisting of 0.03% acetic acid (v/v) and methanol at a flow rate of 0.50 mL/min. The linearity, recovery, matrix effect, accuracy, precision, and stability of the method were well validated. We evaluated drug–drug interactions based on these four compounds in freshly suspended hepatocytes. The hepatic uptake of pravastatin, fexofenadine, rosuvastatin, and methotrexate at 4°C was significantly lower than that at 37°C, and the hepatocytes were saturable with increased substrate concentration and culture time, suggesting that the rat primary hepatocyte model was successfully established. Triptolide showed a significant inhibitory effect on the hepatic uptake of these four compounds. In conclusion, this method was successfully employed for the quantification of pravastatin, fexofenadine, rosuvastatin, and methotrexate and was used to verify the rat primary hepatocyte model for Oatp1, Oatp2, Oatp4, and Oat2 transporter studies. Then, we applied this model to explore the effect of triptolide on these four transporters.