Chavan Balasaheb B, Kalariya Pradipbhai D, Nimbalkar Rakesh D, Garg Prabha, Srinivas R, Kumar Talluri M V N
Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education & Research, IDPL R&D Campus, Balanagar, Hyderabad, 500 037, India.
Department of Pharmacoinformatics, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar, Punjab, 160 062, India.
J Mass Spectrom. 2017 May;52(5):296-314. doi: 10.1002/jms.3929.
The present study reports the in vivo and in vitro identification and characterization of metabolites of fluvastatin, the 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase inhibitor, using liquid chromatography-mass spectrometry (LC-MS). In vitro studies were conducted by incubating the drug with human liver microsomes and rat liver microsomes. In vivo studies were carried out by administration of the drug in the form of suspension to the Sprague-Dawley rats followed by collection of urine, faeces and blood at different time points up to 24 h. Further, samples were prepared by optimized sample preparation method, which includes freeze liquid extraction, protein precipitation and solid phase extraction. The extracted and concentrated samples were analysed using ultrahigh-performance liquid chromatography-quadruple time-of-flight tandem mass spectrometry. A total of 15 metabolites were observed in urine, which includes hydroxyl, sulphated, desisopropyl, dehydrogenated, dehydroxylated and glucuronide metabolites. A few of the metabolites were also present in faeces and plasma samples. In in vitro studies, a few metabolites were observed that were also present in in vivo samples. All the metabolites were characterized using ultrahigh-performance liquid chromatography-quadruple time-of-flight tandem mass spectrometry in combination with accurate mass measurement. Finally, in silico toxicity studies indicated that some of the metabolites show or possess carcinogenicity and skin sensitization. Several metabolites that were identified in rats are proposed to have toxicological significance on the basis of in silico evaluation. However, these metabolites are of no human relevance. Copyright © 2017 John Wiley & Sons, Ltd.
本研究报告了使用液相色谱 - 质谱联用（LC - MS）对氟伐他汀（一种3 - 羟基 - 3 - 甲基 - 戊二酰辅酶A还原酶抑制剂）的体内和体外代谢产物进行的鉴定和表征。体外研究通过将药物与人肝微粒体和大鼠肝微粒体孵育进行。体内研究通过将药物以悬浮液形式给予Sprague - Dawley大鼠，然后在长达24小时的不同时间点收集尿液、粪便和血液来进行。此外，通过优化的样品制备方法制备样品，该方法包括冷冻液体萃取、蛋白质沉淀和固相萃取。使用超高效液相色谱 - 四极杆飞行时间串联质谱对提取和浓缩后的样品进行分析。在尿液中总共观察到15种代谢产物，包括羟基、硫酸化、去异丙基、脱氢、脱羟基和葡萄糖醛酸代谢产物。其中一些代谢产物也存在于粪便和血浆样品中。在体外研究中，观察到一些代谢产物也存在于体内样品中。所有代谢产物均使用超高效液相色谱 - 四极杆飞行时间串联质谱结合精确质量测量进行表征。最后，计算机模拟毒性研究表明，一些代谢产物显示或具有致癌性和皮肤致敏性。根据计算机模拟评估，在大鼠中鉴定出的几种代谢产物被认为具有毒理学意义。然而，这些代谢产物与人类无关。版权所有© 2017 John Wiley & Sons, Ltd.
