Department of Medical Oncology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands.
Department of Medical Oncology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands.
J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Apr 15;1113:37-44. doi: 10.1016/j.jchromb.2019.03.011. Epub 2019 Mar 11.
The development and full validation of a sensitive and selective ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method are described for the simultaneous analysis of afatinib, alectinib, crizotinib and osimertinib in human lithium heparinized plasma. Afatinib-d6, crizotinib-d5 and erlotinib-d6 were used as internal standards. Given osimertinib's instability in plasma and whole blood at ambient temperature, samples should be solely processed on ice (T = 0 °C). Chromatographic separation was obtained on an Acquity UPLC ® BEH C18; 2.1 × 50 mm, 1.7 μm column, which was eluted with 0.400 mL/minute flow on a linear gradient, consisting of 10 mM ammonium formate (pH 4.5) and acetonitrile. Calibration curves for all compounds were linear for concentration ranges of 1.00 to 100 ng/mL for afatinib and 10.0 to 1000 ng/mL for alectinib, crizotinib and osimertinib, herewith validating the lower limits of quantification at 1.00 ng/mL for afatinib and 10.0 ng/mL for alectinib, crizotinib and osimertinib. Within-run and between-run precision measurements fell within 10.2%, with accuracy ranging from 89.2 to 110%.
描述了一种灵敏且选择性的超高效液相色谱/串联质谱（UPLC-MS/MS）方法的开发和全面验证，用于同时分析人锂肝素化血浆中的阿法替尼、阿来替尼、克唑替尼和奥希替尼。阿法替尼-d6、克唑替尼-d5 和厄洛替尼-d6 被用作内标。鉴于奥希替尼在环境温度下的血浆和全血中的不稳定性，样品应仅在冰上（T=0°C）处理。色谱分离在 Acquity UPLC®BEH C18；2.1×50mm，1.7μm 柱上进行，以 0.400mL/min 的流速在线性梯度洗脱，梯度由 10mM 甲酸铵（pH4.5）和乙腈组成。所有化合物的校准曲线在阿法替尼的浓度范围为 1.00 至 100ng/mL 和阿来替尼、克唑替尼和奥希替尼的浓度范围为 10.0 至 1000ng/mL 时均呈线性，在此验证了阿法替尼的定量下限为 1.00ng/mL，阿来替尼、克唑替尼和奥希替尼的定量下限为 10.0ng/mL。批内和批间精密度测量值均在 10.2%以内，准确度范围为 89.2%至 110%。
