A liquid chromatography–tandem mass spectrometry method was developed and validated to quantify alectinib, crizotinib, erlotinib and gefitinib. This assay can be combined with our method for osimertinib, allowing quantification of the most used ALK- and EGFR-tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer with a single-assay setup. Chromatographic separation was performed on a HyPurity® C₁₈ analytical column using an elution gradient of ammonium acetate in water and in methanol, both acidified with formic acid 0.1%. Detection and quantification were performed using a triple quad mass spectrometer with an electrospray ionization interface. This method led to robust results, as the selectivity, carryover, precision and accuracy met all pre-specified requirements. The assay was validated over a linear range of 100–2,000 ng/ml for alectinib and erlotinib and 50–1,000 ng/ml for crizotinib and gefitinib. Alectinib, crizotinib, erlotinib and gefitinib were all stable for at least 4 h in whole blood (at room temperature and at 4°C) and for at least 1 month in EDTA plasma when stored at −80°C, while osimertinib proved to be unstable at room temperature. Although high-performance liquid chromatography was used, the run time was short and comparable with other methods using ultra-high performance liquid chromatography.

中文翻译：

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开发和验证 HPLC-MS/MS 方法，使用一次检测同时定量人血浆样品中的艾乐替尼、克唑替尼、厄洛替尼、吉非替尼和奥希替尼

开发并验证了液相色谱-串联质谱法，用于定量艾乐替尼、克唑替尼、厄洛替尼和吉非替尼。该测定可以与我们的奥希替尼方法结合使用，从而可以通过单次测定设置对非小细胞肺癌中最常用的ALK和EGFR酪氨酸激酶抑制剂 (TKI)进行量化。在 HyPurity® C ₁₈上进行色谱分离分析柱使用醋酸铵在水和甲醇中的洗脱梯度，两者都用 0.1% 的甲酸酸化。使用具有电喷雾电离接口的三重四极杆质谱仪进行检测和定量。由于选择性、交叉污染、精密度和准确度满足所有预先指定的要求，因此该方法产生了稳健的结果。该测定在艾乐替尼和厄洛替尼的 100–2,000 ng/ml 线性范围以及克唑替尼和吉非替尼的 50–1,000 ng/ml 线性范围内得到验证。艾乐替尼、克唑替尼、厄洛替尼和吉非替尼在全血中至少稳定 4 小时（室温和 4°C），在 -80°C 保存时在 EDTA 血浆中至少稳定 1 个月，而奥希替尼被证明是在室温下不稳定。虽然使用了高效液相色谱法，