Abstract：
Small molecule inhibitors (SMIs) are increasingly being used in the treatment of non-small cell lung cancer. To support pharmacokinetic research and clinical treatment monitoring, our aim was to develop and validate an ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) assay for quantification of eight SMIs: adagrasib, alectinib, brigatinib, capmatinib, crizotinib, lorlatinib, selpercatinib, and sotorasib. Development of the UPLC-MS/MS assay was done by trying different columns and eluents to optimize peak shape. The assay was validated based on guidelines of the European Medicines Agency. Chromatographic separation was performed with a gradient elution using ammonium formate in water and methanol. Detection was performed using a triple quadrupole tandem mass spectrometer with electrospray ionization. Validation was performed in a range of 10-2500 μg/L for lorlatinib, 25-6250 μg/L for alectinib and crizotinib, 25-10,000 μg/L for capmatinib and selpercatinib, 50-12,500 μg/L for brigatinib, and 100-25,000 μg/L for adagrasib and sotorasib. Imprecision was <8.88% and inaccuracy was <12.5% for all compounds. Seven out of eight compounds were stable for 96 h at room temperature. Sotorasib was stable for 8 h at room temperature. A sensitive and reliable method has been developed to quantify eight SMIs with a single assay, enhancing efficacy and safety of targeted therapies.
摘要：
小分子抑制剂(SMI)越来越多地用于非小细胞肺癌的治疗。为了支持药代动力学研究和临床治疗监测，我们的目标是开发和验证一种超高效液相色谱-质谱（UPLC-MS/MS）测定，用于定量八个SMI：adagrasib，阿列替尼,布加替尼,卡马替尼,克唑替尼,洛拉替尼,selpercatinib,还有Sotorasib.通过尝试不同的柱和洗脱液以优化峰形状来进行UPLC-MS/MS测定的开发。该测定基于欧洲药品管理局的指南进行验证。使用甲酸铵在水和甲醇中的梯度洗脱进行色谱分离。使用具有电喷雾电离的三重四极杆串联质谱仪进行检测。氯拉替尼在10-2500μg/L的范围内进行验证，阿来替尼和克唑替尼为25-6250μg/L，25-10,000μg/L的卡马替尼和selpercatinib，50-12,500μg/L的布格替尼，和100-25,000μg/L的阿达格拉西布和索托拉西布。对于所有化合物,不精确性<8.88%并且不精确性<12.5%。8个化合物中的7个在室温下稳定96小时。Sotorasib在室温下稳定8小时。已开发出一种灵敏可靠的方法，可通过一次测定来量化八个SMI，提高靶向治疗的疗效和安全性。
