Jia Mengqi, Wu Zhenghua, Shi Wenqing, Wang Shuowen, Huang Xucong, Zhang Min, Bao Wuping, Bao Aihua, Zhang Pengyu, Ding Fengming, Xie Guogang, Lou Yuefen, Fan Guorong
Department of Clinical Pharmacy, Shanghai General Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200080, PR China.
Department of Clinical Pharmacy, Shanghai General Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200080, PR China; School of Pharmacy, Shanghai Jiao Tong University, Shanghai, PR China.
J Pharm Biomed Anal. 2023 May 10;228:115275. doi: 10.1016/j.jpba.2023.115275. Epub 2023 Jan 26.
Gefitinib, osimertinib and icotinib are the most commonly used tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC) with EGFR mutation. Therapeutic drug monitoring (TDM) for these TKIs has become a standard and essential procedure. Dried plasma spots (DPS) was choosen for microsampling strategies for TDM, allowing easy and cost-effective logistics in many settings. This study developd and validated an assay for the simultaneous quantitative determination of gefitinib, osimertinib and icotinib in DPS by online solid-phase extraction-liquid chromatography-tandem mass spectrometry (online SPE-LC-MS) system. The TKIs were extracted from DPS with methanol and enriched on a Welch Polar-RP SPE column (30 × 4.6 mm, 5 µm), followed by separation on Waters X Bridge C18 analytical column(4.6 × 100 mm, 3.5 µm). The method achieved LLOQ of 2 ng mL for gefitinib and osimertinib (4 ng mL for icotinib), respectively (r > 0.99). Precision (within-run 1.54-7.41 % RSD; between-run 3.03-12.84 % RSD), accuracy (range from 81.47 % to 105.08 %; between-run bias 87.87-104.13 %). Osimertinib and icotinib were stable in DPS stored at - 40 °C for 30 days, 4 °C, 42 °C and 60 °C for 5 days and well-sealed 37 °C,75 % humidity (except gefitinib). Lastly, the assay was applied to TDM of TKIs in 46 patients and the results were compared to SALLE assisted LC-MS analysis, it could be confirmed that the developed method achieves similarly good results as the already established one and no bias could be detected. It implies that this method capable of supporting clinical follow-up TDM of TKIs in DPS from poor medical environment.
吉非替尼、奥希替尼和埃克替尼是治疗表皮生长因子受体（EGFR）突变的非小细胞肺癌（NSCLC）最常用的酪氨酸激酶抑制剂（TKIs）。对这些TKIs进行治疗药物监测（TDM）已成为一项标准且必不可少的程序。干血斑（DPS）被选作TDM的微量采样策略，在许多情况下便于操作且成本效益高。本研究建立并验证了一种在线固相萃取-液相色谱-串联质谱（online SPE-LC-MS）系统同时定量测定DPS中吉非替尼、奥希替尼和埃克替尼的方法。TKIs用甲醇从DPS中提取，并在Welch Polar-RP固相萃取柱（30×4.6 mm，5 µm）上富集，然后在Waters X Bridge C18分析柱（4.6×100 mm，3.5 µm）上分离。该方法对吉非替尼和奥希替尼的定量下限（LLOQ）分别为2 ng/mL（埃克替尼为4 ng/mL）（r>0.99）。精密度（批内相对标准偏差1.54 - 7.41%；批间相对标准偏差3.03 - 12.84%），准确度（范围为81.47%至105.08%；批间偏差87.87 - 104.13%）。奥希替尼和埃克替尼在-40℃储存30天、4℃、42℃和60℃储存5天以及密封状态下37℃、75%湿度（吉非替尼除外）的DPS中均稳定。最后，该方法应用于46例患者的TKIs的TDM，并将结果与SALLE辅助LC-MS分析进行比较，可以确认所建立的方法与已建立的方法取得了相似的良好结果，且未检测到偏差。这意味着该方法能够支持医疗条件较差地区DPS中TKIs的临床随访TDM。
