Yu Lixiu, Zhou Ying, He Xiaomeng, Li Huqun, Chen Hui, Li Weiyong
Institute of Clinical Pharmacy, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, PR China.
Department of Infectious Disease, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2015 Oct 1;1002:358-63. doi: 10.1016/j.jchromb.2015.07.053. Epub 2015 Aug 30.
Macitentan is a newly approved endothelin receptor antagonist (ERA) for the long-term treatment of PAH with superior receptor-binding properties and a longer duration of action compared to other available ERAs. However, analytical methods for simultaneous determination of macitentan and its active metabolite, ACT-132577, in human plasma have not been fully reported in the literature. In this work, a fast, sensitive, and reliable high-performance liquid chromatography-tandem mass spectrometry method (HPLC-MS/MS) was firstly developed and completely validated for simultaneous determination of macitentan and its active metabolite in human plasma. Plasma samples were processed with a protein precipitation using acetonitrile, followed by chromatographic separation using an Inertsil ODS-SP column (100×2.1mm, 3.5μm) under isocratic elution with a mobile phase consisting of acetonitrile and 0.2% formic acid at a flow rate of 0.3mL/min. Quantification was operated in multiple reaction monitoring (MRM) mode using the transitions m/z 547.1→201.0 for macitentan, m/z 589.0→203.0 for ACT-132577, and m/z 380.5→243.3 for the IS (donepezil). The assay exhibited a linear range of 1-500ng/mL for both macitentan and ACT-132577. The accuracy and the intra- and inter-precisions were within acceptable ranges and no significant matrix effect was observed during the method validation. The developed method was successfully utilized to a human pharmacokinetic study of macitentan as well as ACT-132577 after oral administration of 10mg macitentan tablet in healthy Chinese volunteers.
马昔腾坦是一种新获批的内皮素受体拮抗剂（ERA），用于肺动脉高压（PAH）的长期治疗，与其他现有ERA相比，具有更优异的受体结合特性和更长的作用持续时间。然而，文献中尚未充分报道同时测定人血浆中马昔腾坦及其活性代谢物ACT - 132577的分析方法。在本研究中，首次开发并完全验证了一种快速、灵敏且可靠的高效液相色谱 - 串联质谱法（HPLC - MS/MS），用于同时测定人血浆中的马昔腾坦及其活性代谢物。血浆样品采用乙腈进行蛋白沉淀处理，随后使用Inertsil ODS - SP柱（100×2.1mm，3.5μm）在等度洗脱条件下进行色谱分离，流动相由乙腈和0.2%甲酸组成，流速为0.3mL/min。采用多反应监测（MRM）模式进行定量分析，马昔腾坦的质荷比转换为m/z 547.1→201.0，ACT - 132577的质荷比转换为m/z 589.0→203.0，内标（多奈哌齐）的质荷比转换为m/z 380.5→243.3。该测定方法对马昔腾坦和ACT - 132577的线性范围均为1 - 500ng/mL。在方法验证过程中，准确度以及批内和批间精密度均在可接受范围内，且未观察到明显的基质效应。所开发的方法成功应用于健康中国志愿者口服10mg马昔腾坦片后马昔腾坦以及ACT - 132577的人体药代动力学研究。
