Liu Yan, Li Xiang, Yang Chunjuan, Tai Sheng, Zhang Xiangning, Liu Gaofeng
Department of Pharmacy, the Second Affiliated Hospital, Harbin Medical University, Harbin 150086, China.
J Chromatogr Sci. 2013 Jan;51(1):26-32. doi: 10.1093/chromsci/bms100. Epub 2012 Jun 12.
A specific ultra-performance liquid chromatography tandem mass spectrometry method has been described for the simultaneous determination of caffeine, tolbutamide, metoprolol and dapsone in rat plasma, which are the four probe drugs of the four cytochrome P450 (CYP450) isoforms CYP1A2, CYP2C9, CYP2D6 and CYP3A4. The chromatographic separation was achieved using a Waters Acquity UPLC BEH HILIC C(18) column (2.1 × 50 mm, 1.7 µm). The mobile phase consisted of acetonitrile and water (containing 0.1% formic acid) (15:85, v/v). The triple quadrupole mass spectrometric detection was operated by positive electrospray ionization. Phenacetin was chosen as internal standard. Plasma samples were extracted with dichloromethane-butanol (10:1, v/v). The recoveries ranged from 67.5% to 98.5%. The calibration curves in plasma were linear in the range of 2.5-1,000 ng/mL for caffeine and dapsone, 5-5,000 ng/mL for tolbutamide and 2.5-250 ng/mLfor metoprolol, with correlation coefficient (r(2)) of 0.9936, 0.9966, 0.9990 and 0.9998, respectively. The method was successfully applied to pharmacokinetic studies of the four probe drugs of the four CYP450 isoforms and used to evaluate the effects of breviscapine on the activities of CYP1A2, CYP2C9, CYP2D6 and CYP3A4 in rats.
已描述了一种特定的超高效液相色谱串联质谱法，用于同时测定大鼠血浆中的咖啡因、甲苯磺丁脲、美托洛尔和氨苯砜，这四种药物是细胞色素P450（CYP450）四种同工酶CYP1A2、CYP2C9、CYP2D6和CYP3A4的四种探针药物。采用沃特世Acquity UPLC BEH HILIC C18柱（2.1×50 mm，1.7 µm）进行色谱分离。流动相由乙腈和水（含0.1%甲酸）（15:85，v/v）组成。采用正电喷雾电离进行三重四极杆质谱检测。选择非那西丁作为内标。血浆样品用二氯甲烷 - 丁醇（10:1，v/v）萃取。回收率在67.5%至98.5%之间。血浆中的校准曲线在咖啡因和氨苯砜为2.5 - 1000 ng/mL、甲苯磺丁脲为5 - 5000 ng/mL、美托洛尔为2.5 - 250 ng/mL范围内呈线性，相关系数（r2）分别为0.9936、0.9966、0.9990和0.9998。该方法成功应用于四种CYP450同工酶的四种探针药物的药代动力学研究，并用于评估灯盏花素对大鼠CYP1A2、CYP2C9、CYP2D6和CYP3A4活性的影响。
