Retmana Irene A, Loos Nancy H C, Schinkel Alfred H, Beijnen Jos H, Sparidans Rolf W
Division of Pharmacology, Department of Pharmaceutical Sciences, Faculty of Science, Utrecht University, Utrecht, The Netherlands.
Division of Pharmacology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Biomed Chromatogr. 2023 Nov;37(11):e5720. doi: 10.1002/bmc.5720. Epub 2023 Aug 19.
We developed and validated an assay utilizing a liquid chromatography-tandem mass spectrometry technique to quantify the KRAS inhibitor adagrasib in mouse plasma and seven tissue-related matrices. The straightforward protein precipitation technique was selected to extract adagrasib and the internal standard salinomycin from the matrices. Gradient elution of acetonitrile and water modified with 0.5% (v/v) ammonium hydroxide and 0.02% (v/v) acetic acid on a C column at a flow rate of 0.6 ml/min was applied to separate the analytes. Both adagrasib and salinomycin were detected with a triple quadrupole mass spectrometer with positive electrospray ionization in a selected reaction monitoring mode. A linear calibration range of 2-2,000 ng/ml of adagrasib was demonstrated during the validation. In addition, the reported precision values (intra- and inter-day) were between 3.5 and 14.9%, while the accuracy values were 85.5-111.0% for all tested levels in all investigated matrices. Adagrasib in mouse plasma was reported to have good stability at room temperature, while adagrasib in tissue-related matrices was stable on ice for up to 4 h (matrix dependent). Finally, this method was successfully applied to determine the pharmacokinetic profile and tissue distribution of adagrasib in wild-type mice.
我们开发并验证了一种利用液相色谱 - 串联质谱技术来定量测定小鼠血浆和七种组织相关基质中KRAS抑制剂阿达格拉西布的分析方法。选用简单的蛋白沉淀技术从基质中提取阿达格拉西布和内标沙利霉素。在C柱上，以0.6 ml/min的流速，用0.5%（v/v）氢氧化铵和0.02%（v/v）乙酸改性的乙腈和水进行梯度洗脱，以分离分析物。阿达格拉西布和沙利霉素均采用三重四极杆质谱仪，在正电喷雾电离的选择反应监测模式下进行检测。在验证过程中，阿达格拉西布的线性校准范围为2 - 2,000 ng/ml。此外，在所有研究基质的所有测试水平下，报告的精密度值（日内和日间）在3.5%至14.9%之间，而准确度值为85.5% - 111.0%。据报道，小鼠血浆中的阿达格拉西布在室温下具有良好的稳定性，而组织相关基质中的阿达格拉西布在冰上最多可稳定4小时（取决于基质）。最后，该方法成功应用于测定野生型小鼠中阿达格拉西布的药代动力学特征和组织分布。
