Department of Pharmacy, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Department of Oncology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Oct 1;1182:122940. doi: 10.1016/j.jchromb.2021.122940. Epub 2021 Sep 17.
Dacomitinib, an irreversible pan-ErbB tyrosine kinase inhibitor targeting the human epidermal growth factor receptor, is used for the treatment of metastatic non-small cell lung cancer. To facilitate the investigations on its metabolism and other relevant studies, based on high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), a rapid and sensitive bioanalytical technique was established and fully validated for simultaneous quantification of dacomitinib and its main metabolite in human plasma. The plasma samples were treated with acetonitrile containing 0.1% formic acid and the liquid supernatant was collected, dried and dissolved in methanol-water-formic acid (200:800:1, v/v) before injection. The chromatographic separation was performed on an ACE Excel C column (2.1 mm × 50.0 mm, i.d., 5 μm) by gradient elution with a mixture of buffer (5 mM ammonium acetate in 0.1% formic acid) and acetonitrile, serving as the mobile phase, with an overall run time of 4 min. Dacomitinib, O-desmethyl dacomitinib and IS were subsequently detected on an AB QTRAP 5500 mass spectrometer in positive ion and multiple reaction monitoring modes at the precursor-to-product transitions of m/z 470.4 → 385.0, m/z 456.0 → 370.9 and m/z 480.2 → 385.1, respectively. The accuracy and precision of determinations were guaranteed within the concentration ranges of 0.25-100 ng/mL for dacomitinib and 0.20-80 ng/mL for O-desmethyl dacomitinib. The intra- and inter-assay accuracy ranged from 92.00% to 104.50% and the intra- and inter-assay precision was less than 8.20% for each analyte. The method was validated and the relevant parameters, including selectivity, interference among analytes and internal standard, carry-over effect, dilution integrity, extraction recovery, matrix effect, and stability, all satisfied the requirements formulated by the US Food and Drug Administration and the European Medicines Agency. The clinical applicability of the fully-validated method was evaluated in medicated samples from patients on dacomitinib.
达可替尼是一种不可逆的泛 ErbB 酪氨酸激酶抑制剂，针对人表皮生长因子受体，用于治疗转移性非小细胞肺癌。为了促进其代谢等相关研究，基于高效液相色谱-串联质谱法（HPLC-MS/MS），建立并充分验证了一种同时定量测定人血浆中达可替尼及其主要代谢物的快速灵敏的生物分析方法。血浆样品用含 0.1%甲酸的乙腈处理，收集上清液，干燥后用甲醇-水-甲酸（200:800:1，v/v）溶解，然后进样。色谱分离在 ACE Excel C 柱（2.1mm×50.0mm，内径，5μm）上进行，采用缓冲液（0.1%甲酸中的 5mM 乙酸铵）和乙腈的混合液作为流动相进行梯度洗脱，总运行时间为 4min。达可替尼、O-去甲基达可替尼和 IS 随后在 AB QTRAP 5500 质谱仪上以正离子和多重反应监测模式检测，在 m/z 470.4→385.0、m/z 456.0→370.9 和 m/z 480.2→385.1 处分别检测到母离子到产物离子的跃迁。达可替尼的测定浓度范围为 0.25-100ng/mL，O-去甲基达可替尼的测定浓度范围为 0.20-80ng/mL，测定的准确性和精密度均在该范围内得到保证。每个分析物的日内和日间准确度在 92.00%至 104.50%之间，日内和日间精密度均小于 8.20%。该方法经过验证，相关参数，包括选择性、分析物和内标之间的干扰、交叉污染效应、稀释完整性、提取回收率、基质效应和稳定性，均符合美国食品和药物管理局和欧洲药品管理局制定的要求。在接受达可替尼治疗的患者的用药样本中评估了完全验证方法的临床适用性。
