Utrecht University, Faculty of Science, Department of Pharmaceutical Sciences, Division of Pharmacoepidemiology & Clinical Pharmacology, Universiteitsweg 99, 3584 CG Utrecht, The Netherlands; Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, Polifunzionale, 87036 Rende, Italy.
The Netherlands Cancer Institute, Division of Pharmacology, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Apr 15;1083:204-208. doi: 10.1016/j.jchromb.2018.03.014. Epub 2018 Mar 9.
A bio-analytical assay for the first third generation ALK inhibitor lorlatinib in mouse plasma was developed and validated. Ten-μl plasma samples were prepared by adding rucaparib as the internal standard and precipitation of the plasma proteins. For LC-MS/MS analysis, compounds were eluted at 0.5 mL/min and separated on a 3-μm particle-size, polar embedded octadecyl silica column by gradient elution using 0.1% of formic acid (in water) and methanol. Compounds were monitored with positive electrospray ionization using a triple quadrupole mass spectrometer in selected reaction monitoring mode. The assay was fully validated in the 2-2000 ng/mL calibration range. Within-day (8.0-11.6%) and between-day (10.0-15.0%) precisions and accuracies (99.0-113.3%) were within acceptable range. Plasma samples were deemed stable for 6 h at ambient temperature, during three freeze-thaw cycles and for 2 months at -30 °C. Finally, the new assay was applied successfully to pilot pharmacokinetic studies in male and female wild-type mice.
建立并验证了一种用于检测第一代三代 ALK 抑制剂洛拉替尼在小鼠血浆中的生物分析检测方法。采用鲁卡帕尼（内标）沉淀蛋白，10μL 血浆样品经处理后，以 0.5mL/min 的流速，在 3μm 粒径、极性嵌入十八烷基硅烷柱上，通过 0.1%甲酸（水中）-甲醇梯度洗脱进行 LC-MS/MS 分析。采用正电喷雾电离，在选择反应监测模式下，用三重四极杆质谱仪进行检测。该方法在 2-2000ng/mL 的校准范围内得到了全面验证。日内（8.0-11.6%）和日间（10.0-15.0%）精密度和准确度（99.0-113.3%）均在可接受范围内。在环境温度下，血浆样品稳定 6 小时，经过三个冻融循环和在-30°C 下保存 2 个月。最后，该新方法成功应用于雄性和雌性野生型小鼠的初步药代动力学研究。
