Bosilkovska Marija, Déglon Julien, Samer Caroline, Walder Bernhard, Desmeules Jules, Staub Christian, Daali Youssef
Division of Clinical Pharmacology & Toxicology, Geneva University Hospitals, Rue Gabrielle Perret-Gentil 4, 1211, Geneva, Switzerland.
Bioanalysis. 2014 Jan;6(2):151-64. doi: 10.4155/bio.13.289.
An LC-MS/MS method has been developed for the simultaneous quantification of P-glycoprotein (P-gp) and cytochrome P450 (CYP) probe substrates and their Phase I metabolites in DBS and plasma. P-gp (fexofenadine) and CYP-specific substrates (caffeine for CYP1A2, bupropion for CYP2B6, flurbiprofen for CYP2C9, omeprazole for CYP2C19, dextromethorphan for CYP2D6 and midazolam for CYP3A4) and their metabolites were extracted from DBS (10 µl) using methanol. Analytes were separated on a reversed-phase LC column followed by SRM detection within a 6 min run time.
The method was fully validated over the expected clinical concentration range for all substances tested, in both DBS and plasma. The method has been successfully applied to a PK study where healthy male volunteers received a low dose cocktail of the here described P-gp and CYP probes. Good correlation was observed between capillary DBS and venous plasma drug concentrations.
Due to its low-invasiveness, simple sample collection and minimal sample preparation, DBS represents a suitable method to simultaneously monitor in vivo activities of P-gp and CYP.
已开发出一种液相色谱-串联质谱（LC-MS/MS）方法，用于同时定量干血斑（DBS）和血浆中P-糖蛋白（P-gp）及细胞色素P450（CYP）探针底物及其I相代谢产物。使用甲醇从10 μl DBS中提取P-gp（非索非那定）和CYP特异性底物（CYP1A2的咖啡因、CYP2B6的安非他酮、CYP2C9的氟比洛芬、CYP2C19的奥美拉唑、CYP2D6的右美沙芬和CYP3A4的咪达唑仑）及其代谢产物。在反相液相色谱柱上分离分析物，然后在6分钟的运行时间内进行选择反应监测（SRM）检测。
该方法在DBS和血浆中所有测试物质的预期临床浓度范围内均得到充分验证。该方法已成功应用于一项药代动力学（PK）研究，健康男性志愿者接受了此处所述的P-gp和CYP探针的低剂量混合药物。观察到毛细血管DBS与静脉血浆药物浓度之间具有良好的相关性。
由于其侵入性低、样本采集简单且样本制备最少，DBS是一种同时监测P-gp和CYP体内活性的合适方法。
