Clinical Pharmacy Center, Department of Pharmacy, Zhejiang Provincial People's Hospital, Affiliated People's Hospital, Hangzhou Medical College, Hangzhou, Zhejiang, China; College of pharmacy, Zhejiang University of Technology, Hangzhou 310014, China.
Clinical Pharmacy Center, Department of Pharmacy, Zhejiang Provincial People's Hospital, Affiliated People's Hospital, Hangzhou Medical College, Hangzhou, Zhejiang, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2022 Oct 1;1208:123394. doi: 10.1016/j.jchromb.2022.123394. Epub 2022 Jul 28.
The objective of this study was to develop and validate a simple, rapid, and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of three tyrosine kinase inhibitors (ceritinib, osimertinib, and crizotinib) in human plasma using a single-step protein precipitation extraction. Chromatographic separation was achieved using a Waters X Bridge C18 (2.1 mm × 100 mm, 3.5 µm) and gradient elution with 0.2 % formic acid in water and acetonitrile. The total run time was 4.0 min, and the injection volume was 5 μL. The analytes were detected in the multiple reaction monitoring mode using electrospray ionization with positive ion mode. The m/z transitions of ceritinib, osimertinib, crizotinib and nilotinib were 558.0 → 433.2, 500.0 → 72.1, 450.0 → 259.3, and 530.0 → 289.1, respectively. The method was linear in the range of 2-500 ng/mL with lower limit of quantification of 2 ng/mL. Based on the guidelines on bioanalytical methods by the FDA, the validation studies demonstrated that the three analytes were both precise and accurate at four concentration levels, and the coefficient of variation was < 10.59 % and accuracy was > 88.26 %. We present a simple, rapid, and sensitive method for the simultaneous quantification of ceritinib, osimertinib, and crizotinib in human plasma by LC-MS/MS, which could be used in routine therapeutic drug monitoring.
本研究旨在开发并验证一种简单、快速和灵敏的液相色谱-串联质谱（LC-MS/MS）方法，用于在单次蛋白沉淀提取的基础上，同时测定人血浆中的三种酪氨酸激酶抑制剂（塞瑞替尼、奥希替尼和克唑替尼）。采用 Waters X Bridge C18（2.1mm×100mm，3.5μm）色谱柱，以 0.2%甲酸水溶液和乙腈为流动相进行梯度洗脱，实现了色谱分离。总运行时间为 4.0 分钟，进样量为 5μL。采用正离子模式电喷雾电离，多反应监测模式进行检测。塞瑞替尼、奥希替尼、克唑替尼和尼罗替尼的 m/z 跃迁分别为 558.0→433.2、500.0→72.1、450.0→259.3 和 530.0→289.1。该方法在 2-500ng/mL 范围内呈线性，定量下限为 2ng/mL。根据 FDA 生物分析方法指南，验证研究表明，三种分析物在四个浓度水平下均具有良好的精密度和准确度，变异系数<10.59%，准确度>88.26%。本研究建立了一种简单、快速、灵敏的 LC-MS/MS 同时测定人血浆中塞瑞替尼、奥希替尼和克唑替尼的方法，可用于常规治疗药物监测。
