Zheng Xin, Wang Weicong, Zhang Yanbao, Ma Yuxiang, Zhao Hongyun, Hu Pei, Jiang Ji
Clinical Pharmacology Research Center, Peking Union Medical College Hospital, China.
Department of Clinical Trial Center, China National Clinical Research Center for Neurological Diseases, Beijing Tiantan Hospital, Capital Medical University, China.
Biomed Chromatogr. 2018 Dec;32(12):e4365. doi: 10.1002/bmc.4365. Epub 2018 Sep 12.
Osimertinib (AZD9291) is a highly selective irreversible epidermal growth factor receptor tyrosine kinase inhibitor (EGFR TKI) designed to treat nonsmall-cell lung cancer patients with EGFR active and T790 M resistant mutations. A rapid and sensitive method for quantitative analysis by ultra-performance liquid chromatography-tandem mass spectrometry of osimertinib and its metabolite AZ5104 in human plasma was developed and validated. The samples were prepared by protein precipitation and separated on a BEH C column (2.1 × 50 mm, 1.7 μm) by gradient elution with 0.1% (v/v) formic acid and 10 mm ammonium acetate in water and acetonitrile as the mobile phase. Electrospray ionization in positive ion mode and multiple reaction monitoring were used to monitor the ion transitions at m/z 500.4 → 385.3 and 486.3 → 371.1. The results indicated that the method had excellent sensitivity and specificity. The validation was performed in a range from 0.5 to 100 ng/mL. Intra-day and inter-day precisions (in terms of RSD) were all <15%, and the accuracies (in terms of RE) were within ±15%. The lower limit of quantification, matrix effect, extraction recovery, stability and dilution integrity were also validated and satisfied the validation criteria. Finally, this method was successfully applied in a retrospective analysis, and the predose samples of 52 nonsmall-cell lung cancer patients who were enrolled in a novel third EGFR TKI clinical trial were analyzed for screening regardless of whether they had previously received osimertinib treatments.
奥希替尼（AZD9291）是一种高度选择性的不可逆表皮生长因子受体酪氨酸激酶抑制剂（EGFR TKI），旨在治疗具有EGFR激活和T790M耐药突变的非小细胞肺癌患者。建立并验证了一种采用超高效液相色谱-串联质谱法对人血浆中奥希替尼及其代谢物AZ5104进行定量分析的快速灵敏方法。样品通过蛋白沉淀法制备，在BEH C柱（2.1×50 mm，1.7μm）上以0.1%（v/v）甲酸和10 mM醋酸铵的水和乙腈混合溶液为流动相进行梯度洗脱分离。采用正离子模式下的电喷雾电离和多反应监测来监测m/z 500.4→385.3和486.3→371.1的离子跃迁。结果表明该方法具有出色的灵敏度和特异性。在0.5至100 ng/mL范围内进行了方法验证。日内和日间精密度（以RSD计）均<15%，准确度（以RE计）在±15%以内。还对定量下限、基质效应、提取回收率、稳定性和稀释完整性进行了验证，均符合验证标准。最后，该方法成功应用于一项回顾性分析，对52例参加新型第三代EGFR TKI临床试验的非小细胞肺癌患者的给药前样本进行了分析筛选，无论他们之前是否接受过奥希替尼治疗。
