Uno Tsukasa, Yasui-Furukori Norio, Shimizu Mikiko, Sugawara Kazunobu, Tateishi Tomonori
Department of Pharmacy, Hirosaki University Hospital, Hirosaki 036-8563, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Sep 25;824(1-2):238-43. doi: 10.1016/j.jchromb.2005.07.027.
A new sensitive column-switching high-performance liquid chromatographic (HPLC) method with ultraviolet detection was developed for the simultaneous determination of rabeprazole, a proton pump inhibitor, and its active metabolite, rabeprazole thioether in human plasma. Rabeprazole, its thioether metabolite and 5-methyl-2-[(4-(3-methoxypropoxy)-3-methyl pyridin-2-yl) methyl sulfinyl]-1H-benzimidazole, as an internal standard were extracted from 1 ml of plasma using diethyl ether-dichloromethane (9:1, v/v) mixture and the extract was injected into a column I (TSK-PW precolumn, 10 microm, 35 mmx4.6mm I.D.) for clean-up and column II (C18 Grand ODS-80TM TS analytical column, 5 microm, 250 mmx4.6 mm I.D.) for separation. The peak was detected with an ultraviolet detector set at a wavelength of 288 nm, and the total time for chromatographic separation was approximately 25 min. Mean absolute recoveries were 78.0 and 88.3% for rabeprazole and rabeprazole thioether, respectively. Intra- and inter-day coefficient variations were less than 6.5 and 4.5% for rabeprazole, 3.6 and 5.3% for rabeprazole thioether, respectively, at the different concentration ranges. The validated concentration ranges of this method were 1-1000 ng/ml for rabeprazole and 3-500 ng/ml for rabeprazole thioether. The limits of quantification were 1 ng/ml for rabeprazole and 3 ng/ml for rabeprazole thioether. The method was suitable for therapeutic drug monitoring and was applied to pharmacokinetic study in human volunteers.
建立了一种新的带有紫外检测的灵敏的柱切换高效液相色谱（HPLC）法，用于同时测定人血浆中的质子泵抑制剂雷贝拉唑及其活性代谢物雷贝拉唑硫醚。雷贝拉唑、其硫醚代谢物以及作为内标的5-甲基-2-[(4-(3-甲氧基丙氧基)-3-甲基吡啶-2-基)甲基亚磺酰基]-1H-苯并咪唑，用乙醚-二氯甲烷（9:1，v/v）混合物从1 ml血浆中萃取，萃取物注入柱I（TSK-PW预柱，10μm，35mm×4.6mm内径）进行净化，再注入柱II（C18 Grand ODS-80TM TS分析柱，5μm，250mm×4.6mm内径）进行分离。用设置在288nm波长的紫外检测器检测峰，色谱分离总时间约为25分钟。雷贝拉唑和雷贝拉唑硫醚的平均绝对回收率分别为78.0%和88.3%。在不同浓度范围内，雷贝拉唑的日内和日间系数变异分别小于6.5%和4.5%，雷贝拉唑硫醚的分别为3.6%和5.3%。该方法的有效浓度范围为雷贝拉唑1-1000 ng/ml，雷贝拉唑硫醚3-500 ng/ml。定量限为雷贝拉唑1 ng/ml，雷贝拉唑硫醚3 ng/ml。该方法适用于治疗药物监测，并应用于人体志愿者的药代动力学研究。
