Department of Clinical Pharmacology, Faculty of Pharmaceutical Sciences, Hokkaido University of Science, Hokkaido, Japan.
Department of Pharmacy, University of Tsukuba Hospital, Ibaraki, Japan.
Ther Drug Monit. 2021 Dec 1;43(6):772-779. doi: 10.1097/FTD.0000000000000895.
Routine therapeutic drug monitoring is a promising approach for the rational use of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) and anaplastic lymphoma kinase (ALK) inhibitors. The purpose of this study was to develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of 5 EGFR-TKIs (afatinib, dacomitinib, erlotinib, gefitinib, and osimertinib) and 3 ALK inhibitors (alectinib, ceritinib, and crizotinib).
A 100-mL aliquot of serum was diluted with 100 μL of 1% aqueous ammonia containing internal standards and then purified using the supported liquid extraction method. LC-MS/MS was conducted in positive ionization mode, and the method was validated according to published guidelines.
Calibration curves were linear across concentration ranges examined. The intra- and interassay accuracies were 90.7%-110.7% and 94.7%-107.6%, respectively. All intra- and interassay imprecision values were ≤10.1%. The EGFR-TKIs and ALK inhibitors examined in this study, except osimertinib, which could be stored on ice for at least 5 hours, were stable at room temperature for 3 hours. For the internal standard-normalized matrix factors, the mean recovery and percent coefficient of variation values ranged between 54%-112% and 1.7%-11.7%, respectively. This method successfully determined serum concentrations of afatinib, alectinib, erlotinib, gefitinib, and osimertinib in clinical samples. Serum levels of kinase inhibitors consistently reflected those reported in previous studies.
An LC-MS/MS method suitable for the simultaneous determination of 5 EGFR-TKIs and 3 ALK inhibitors in serum was developed and validated. The newly developed method enabled the determination of 5 of 8 target drugs examined in clinical samples. However, a large number of clinical samples need to be analyzed to verify the usefulness of the method.
常规治疗药物监测是合理使用表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKIs）和间变性淋巴瘤激酶（ALK）抑制剂的有前途的方法。本研究的目的是开发和验证一种同时测定 5 种 EGFR-TKIs（阿法替尼、达可替尼、厄洛替尼、吉非替尼和奥希替尼）和 3 种 ALK 抑制剂（阿来替尼、塞瑞替尼和克唑替尼）的液相色谱-串联质谱（LC-MS/MS）方法。
取 100ml 血清样本，用 100μL 含内标物的 1%氨水溶液稀释，然后采用固相萃取法进行净化。LC-MS/MS 采用正离子化模式进行，方法根据已发表的指南进行验证。
校准曲线在检测浓度范围内呈线性。日内和日间准确度分别为 90.7%-110.7%和 94.7%-107.6%。所有日内和日间精密度值均≤10.1%。本研究检测的 EGFR-TKIs 和 ALK 抑制剂（奥希替尼除外，奥希替尼可在冰上至少稳定 5 小时）在室温下 3 小时内稳定。对于内标归一化基质因子，平均回收率和变异系数值分别在 54%-112%和 1.7%-11.7%之间。该方法成功测定了临床样本中阿法替尼、阿来替尼、厄洛替尼、吉非替尼和奥希替尼的血清浓度。激酶抑制剂的血清水平与之前研究报道的一致。
开发并验证了一种适用于同时测定血清中 5 种 EGFR-TKIs 和 3 种 ALK 抑制剂的 LC-MS/MS 方法。新开发的方法能够测定 8 种目标药物中的 5 种在临床样本中的浓度。然而，需要分析大量的临床样本来验证该方法的实用性。
