Levent Serkan, Elriş Abeer, Avcı Hazal, Özcan Saniye, Can Nafiz Öncü
Department of Analytical Chemistry, Faculty of Pharmacy, Anadolu University, Yunusemre Campus, 26470, Eskisehir, Turkey.
Central Analysis Laboratory (MERLAB), Faculty of Pharmacy, Anadolu University, 26470, Eskisehir, Turkey.
BMC Chem. 2025 May 28;19(1):147. doi: 10.1186/s13065-025-01509-y.
Alectinib, the most common second-generation anaplastic lymphoma kinase inhibitor, is efficacious against various anaplastic lymphoma kinase mutations and is utilized in the treatment of non-small cell lung cancer. In this work, the determination of both alectinib and its impurity 5-trifluoroacetate at the same time was realized by developing a novel high performance liquid chromatography method. The stationary phase was an Ascentis Express 90 Å C (10 cm × 4.6 mm, 2.7 µm) HPLC column; the separation was obtained using a gradient system with a mobile phase consisting of acetonitrile and ammonium acetate buffer, and the responses were recorded from the PDA detector at a wavelength of 269 nm. The method's optimization was carried out using the Box-Behnken design, whereas the optimized conditions were selected according to the desirability function. Validation of the method was done with the International Council for Harmonization, ICH Q2(R2) Guidelines. The limit of detection values was 0.1 and 0.3 µg/mL, and the limit of quantification values were 0.3 and 0.5 µg/mL for alectinib and its impurity, respectively. The recovery study was realized in the Alecensa capsule (150 mg alectinib/capsule), with high recoveries showing the perfect precision and accuracy of the method. A bottom-up approach was used to estimate the measurement uncertainty. This made it possible to find and measure the sources of uncertainty as well as the factors that affected the chromatographic responses. The resulting regression models were deemed adequate and were then used to define the operable design region using the Monte Carlo method. The eco-friendliness of the proposed methods was validated with the Complex Green Analytical Procedure Index, Analytical Greenness Metric and Blue Applicability Grade Index greenness evaluation tools. Also, the whiteness of the method was calculated with the newly developed White Analytical Chemistry tool. The method can be used to assay alectinib and its impurity agents in its formulation in quality control laboratories.
阿来替尼是最常见的第二代间变性淋巴瘤激酶抑制剂，对各种间变性淋巴瘤激酶突变均有效，用于治疗非小细胞肺癌。在本研究中，通过开发一种新型高效液相色谱法，实现了同时测定阿来替尼及其杂质5-三氟乙酸盐。固定相为Ascentis Express 90 Å C（10 cm×4.6 mm，2.7 µm）高效液相色谱柱；采用梯度洗脱系统进行分离，流动相由乙腈和醋酸铵缓冲液组成，通过PDA检测器在269 nm波长处记录响应信号。采用Box-Behnken设计对方法进行优化，并根据期望函数选择优化条件。按照国际协调理事会（ICH）Q2(R2)指南对该方法进行验证。阿来替尼及其杂质的检测限分别为0.1和0.3 µg/mL，定量限分别为0.3和0.5 µg/mL。回收率研究在Alecensa胶囊（每粒含150 mg阿来替尼）中进行，高回收率表明该方法具有良好的精密度和准确度。采用自下而上的方法估计测量不确定度。这使得能够找到并测量不确定度的来源以及影响色谱响应的因素。所得回归模型被认为是合适的，然后使用蒙特卡罗方法定义可操作设计区域。使用综合绿色分析程序指数、分析绿色度指标和蓝色适用性等级指数等绿色度评估工具对所提出方法的环境友好性进行验证。此外，使用新开发的白色分析化学工具计算该方法的白色度。该方法可用于质量控制实验室中阿来替尼及其制剂中杂质的测定。
