Giri Kalpesh Kumar, Suresh P S, Saim Syed Mohd, Zainuddin Mohd, Bhamidipati Ravi Kanth, Dewang Purushottam, Hallur Mahanandeesha S, Rajagopal Sridharan, Rajagopal Sriram, Mullangi Ramesh
Drug Metabolism and Pharmacokinetics, Jubilant Biosys Ltd, Industrial Suburb, Bangalore, India.
Department of Medicinal Chemistry, Jubilant Biosys Ltd, Industrial Suburb, Bangalore, India.
Biomed Chromatogr. 2017 Jul;31(7). doi: 10.1002/bmc.3912. Epub 2017 Jan 22.
A sensitive and rapid LC-MS/MS method was developed and validated for the simultaneous quantitation of four HDAC inhibitors, namely belinostat (BST), panobinostat (PST), rocilinostat (RST) and vorinostat (VST), in mouse plasma as per regulatory guidelines. The analytes and internal standard were extracted from 50 μL mouse plasma by protein precipitation, followed by chromatographic separation using an Atlantis C column with an isocratic mobile phase comprising 0.1% formic acid-acetonitrile (25:75, v/v) at a flow rate of 0.5 mL/min within 2.5 min. Detection and quantitation were done by multiple reaction monitoring on a triple quadrupole mass spectrometer following the transitions: m/z 319 → 93, 350 → 158, 434 → 274 and 265 → 232 for BST, PST, RST and VST, respectively, in the positive ionization mode. The calibration curves were linear from 2.92 to 2921 ng/mL for BST and PST and from 1.01 to 1008 ng/mL for RST and VST with r ≥ 0.99 for all of the analytes. The intra- and inter-batch accuracy and precision (CV) across quality controls varied from 85.5 to 112% and from 2.30 to 12.5, respectively, for all of the analytes. Analytes were found to be stable under different stability conditions. The method was applied to an i.v. pharmacokinetic study in mice.
按照监管指南，开发并验证了一种灵敏、快速的液相色谱-串联质谱（LC-MS/MS）方法，用于同时定量小鼠血浆中的四种组蛋白去乙酰化酶（HDAC）抑制剂，即贝利司他（BST）、帕比司他（PST）、罗西司他（RST）和伏立诺他（VST）。通过蛋白沉淀从50 μL小鼠血浆中提取分析物和内标，然后使用Atlantis C柱进行色谱分离，等度流动相为含0.1%甲酸的乙腈（25:75，v/v），流速为0.5 mL/min，分离时间为2.5分钟。在三重四极杆质谱仪上通过多反应监测进行检测和定量，在正离子模式下，BST、PST、RST和VST的跃迁分别为：m/z 319→93、350→158、434→274和265→232。BST和PST的校准曲线在2.92至2921 ng/mL范围内呈线性，RST和VST的校准曲线在1.01至1008 ng/mL范围内呈线性，所有分析物的r≥0.99。所有分析物在不同质量控制下的批内和批间准确度和精密度（CV）分别在85.5%至112%和2.30至12.5之间。发现分析物在不同稳定性条件下均稳定。该方法应用于小鼠静脉内药代动力学研究。
