Department of Clinical Pharmacy, Shanghai General Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200080, PR China.
College of Pharmacy, Guangxi University of Chinese Medicine, Nanning 530001, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Jun 15;1175:122752. doi: 10.1016/j.jchromb.2021.122752. Epub 2021 May 1.
In recent years, more than 50 tyrosine kinase inhibitors (TKIs) was indicated against numerous cancers, especially outstanding advantages in the treatment of non-small cell lung cancer (NSCLC), and several studies have shown that therapeutic drug monitoring (TDM) of TKIs can improve treatment efficacy and safety. The present study aimed to develop and validate a LC-MS/MS method for the TDM of 12 TKIs (gefitinib, erlotinib, afatinib, dacomitinib, icotinib, osimertinib, crizotinib, ceritinib, alectinib, dabrafenib, trametinib, anlotinib) in patients with NSCLC. The analytes of interest and internal standard were extracted from human plasma. Salting-out assisted liquid-liquid extraction (SALLE) with 5 M ammonium acetate solution was optimized for method validation and compared to simple protein precipitation (PPT). Chromatographic separation was conducted on Waters X bridge C18 column (100 × 4.6 mm, 3.5 μm) using a gradient elution of acetonitrile/5mM ammonium acetate in pure water with 0.1% (v/v) formic acid at 40 °C within 6 min. The total flow was maintained at 1 mL/min, 30% of the post column flow was split into the mass spectrometer and the rest to waste via a 3-way tee. The mass analysis was performed by positive ion electrospray ionization (ESI) in multiple-reaction monitoring (MRM) mode. The assay was validated based on the guidelines on bioanalytical methods by FDA. This quantification method was proved to be satisfactory in selectivity, accuracy, precision, linearity (r > 0.995), recovery, matrix effect and stability and the accuracy was further assessed in plasma with a degree of hemolysis of 4%. The described method to simultaneously quantify the 12 selected anticancer drugs in human plasma was successfully validated and applied to routine TDM of gefitinib, erlotinib, icotinib, osimertinib, crizotinib and anlotinib in cancer patients. TKIs plasma monitoring helps to individualize dose adjustment and manage adverse effects in NSCLC patients.
近年来，已有 50 多种酪氨酸激酶抑制剂（TKI）被用于治疗多种癌症，尤其是在非小细胞肺癌（NSCLC）的治疗中具有显著优势。几项研究表明，TKI 的治疗药物监测（TDM）可以提高治疗效果和安全性。本研究旨在建立并验证一种 LC-MS/MS 方法，用于测定 NSCLC 患者 12 种 TKI（吉非替尼、厄洛替尼、阿法替尼、达可替尼、埃克替尼、奥希替尼、克唑替尼、色瑞替尼、阿来替尼、达拉非尼、曲美替尼、安罗替尼）的血药浓度。感兴趣的分析物和内标物从人血浆中提取。对盐析辅助液液萃取（SALLE）与 5 M 乙酸铵溶液进行了优化，以用于方法验证，并与简单的蛋白沉淀（PPT）进行了比较。色谱分离在 Waters Xbridge C18 柱（100×4.6mm，3.5μm）上进行，采用乙腈/5mM 乙酸铵在纯水中的梯度洗脱，用 0.1%（v/v）甲酸，40°C 下 6 分钟内洗脱完毕。总流速保持在 1mL/min，30%的柱后分流进入质谱仪，其余部分通过三通分流至废液。采用正离子电喷雾电离（ESI）在多重反应监测（MRM）模式下进行质谱分析。该分析方法基于 FDA 的生物分析方法指南进行验证。该定量方法在选择性、准确性、精密度、线性（r＞0.995）、回收率、基质效应和稳定性方面均表现良好，在 4%溶血度的血浆中进一步评估了准确性。该方法成功验证了同时定量测定人血浆中 12 种选定抗癌药物的方法，并应用于癌症患者吉非替尼、厄洛替尼、埃克替尼、奥希替尼、克唑替尼和安罗替尼的常规 TDM。TKI 血浆监测有助于 NSCLC 患者个体化剂量调整和管理不良反应。
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