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首页> 外文期刊>Journal of Pharmaceutical and Biomedical Analysis: An International Journal on All Drug-Related Topics in Pharmaceutical, Biomedical and Clinical Analysis >Validation of an LC-MS/MS method for simultaneous quantitation of enzalutamide, N-desmethylenzalutamide, apalutamide, darolutamide and ORM-15341 in mice plasma and its application to a mice pharmacokinetic study
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Validation of an LC-MS/MS method for simultaneous quantitation of enzalutamide, N-desmethylenzalutamide, apalutamide, darolutamide and ORM-15341 in mice plasma and its application to a mice pharmacokinetic study

机译：验证对小鼠血浆中苯甲甲酰胺，N-去甲甲醛蛋白质，奥氟丁胺，奥氟酰胺，达尔氟酰胺和ORM-15341的对小鼠药代动力学研究的应用

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摘要

A sensitive and rapid LC-MS/MS method was developed and validated for the simultaneous quantitation of enzalutamide, N-desmethylenzalutamide (active metabolite of enzalutamide), apalutamide, darolu[amide and ORM-15341 (active metabolite of darolutamide) in mice plasma as per regulatory guidelines. The analytes and the internal standard (I.S.: apalutamide-d(3)) were extracted from 50 mu L mice plasma by simple protein precipitation using acetonitrile, followed by chromatographic separation using an Atlantis C-18, column with an isocratic mobile (0.2% formic acid:acetonitrile; 30:70, v/v) at a flow rate of 0.8 mL/min within 2.5 min. Detection and quantitation was done by multiple reaction monitoring on a triple quadrupole mass spectrometer following the transitions: m/z 465 - 209, 451 - 195, 478 - 450, 399 - 178, 397 - 194 and 481 - 453 for enzalutamide, N-desmethylenzalutamide, apalutamide, darolutamide, ORM-15341 and the I.S. respectively. The calibration curves were linear from 1.07 to 2000 ng/mL with r(2) = 0.99 for all the analytes. The intra-and inter-batch accuracy and precision (% CV) across quality controls varied from 88.5-111% and 1.13-13.1, 85.4-106% and 3.15-14.3, respectively for all the analytes. All the analytes were found to be stable under different conditions. The method was applied to an intravenous pharmacokinetic study in mice. (C) 2018 Elsevier B.V. All rights reserved.
机译：开发并验证了敏感和快速的LC-MS / MS方法，用于同时定量苯甲甲酰胺，N-去甲甲醛蛋白质（烯醇丁胺的活性代谢物），奥氟脲酰胺，Darolu [酰胺和ORM-15341（Darolulamide的活性代谢物），如小鼠等离子体每个监管指南。通过使用乙腈的简单蛋白质沉淀从50μl小鼠等离子体中提取分析物和内标（3）），然后使用亚特兰蒂斯C-18，具有等型移动的柱（0.2％）的色谱分离甲酸：乙腈; 30:70，v / v）在2.5分钟内以0.8ml / min的流速。通过在转变后的三重四极杆质谱仪上通过多重反应监测来进行检测和定量：M / Z 465 - ＆GT; 209,451 - ＆ 195,478 - ＆ 450,399 - ＆ 178,397 - ＆ 194和481 - ＆ 453对于烯醇酰胺，N-去甲甲酰胺酰胺，奥氟酰胺，达利氟胺，ORM-15341和I.S。分别。校准曲线从1.07至2000ng / ml线性，所有分析物的R（2）＆ = 0.99。在所有分析物中，跨批量控制的质量控制内和批量准确性和精度（％CV）分别从88.5-111％和1.13-13.1,85.4-106％和3.15-14.3变化。发现所有分析物在不同条件下都被发现是稳定的。将该方法应用于小鼠的静脉内药代动力学研究。 （c）2018年elestvier b.v.保留所有权利。

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