Pharmazentrum Frankfurt/ZAFES, Institut für Klinische Pharmakologie, Klinikum der Johann Wolfgang Goethe-Universität Frankfurt, Theodor-Stern-Kai 7, Haus 74, 60590 Frankfurt am Main, Germany.
J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Dec 1;877(31):4001-6. doi: 10.1016/j.jchromb.2009.10.013. Epub 2009 Oct 14.
The purpose of this study was to develop a specific and highly sensitive method based on fast sample preparation and LC-MS/MS techniques for the determination of the HCV protease inhibitors boceprevir (SCH 503034) and telaprevir (VX 950) in human plasma. Boceprevir, telaprevir and the internal standard dimethylcelecoxib were separated on a Luna C18 column (150 mm x 2.0 mm I.D., 5 microm particle size) under gradient conditions with a mobile phase A consisting of water/ammonia solution (25%) (100:0.05, v/v) and mobile phase B consisting of methanol/ammonia solution (25%) (100:0.05, v/v) and a chromatographic run time of 11 min. The lower limit of quantification (LLOQ) of boceprevir and telaprevir is 0.25 pg on column (25 pg/mL at injection volume of 10 microL). The method possesses a reliable calibration range of 0.025-2.5 ng/mL. Due to the dilution of real life plasma samples by a factor of 10 during the precipitation process the method is suitable to quantify boceprevir and telaprevir at a concentration range of 0.25-25 ng/mL. Variations in accuracy and intraday and interday precision (n=6 for each concentration) were <15% over the whole range of calibration. For the first time, a rapid, specific, sensitive, accurate and reproducible LC-MS/MS method in human plasma has been developed and validated. It is suitable to quantify the concentrations of the hepatitis C virus protease inhibitors boceprevir and telaprevir in human plasma.
本研究旨在开发一种基于快速样品制备和 LC-MS/MS 技术的特异性和高灵敏度方法，用于测定人血浆中的 HCV 蛋白酶抑制剂博赛泼维（SCH503034）和特拉泼维（VX950）。博赛泼维、特拉泼维和内标二甲塞来昔布在 Luna C18 柱（150mm×2.0mmID，5μm 粒径）上，在梯度条件下，以由水/氨溶液（25%）（100：0.05，v/v）组成的流动相 A 和由甲醇/氨溶液（25%）（100：0.05，v/v）组成的流动相 B 洗脱，洗脱时间为 11min。博赛泼维和特拉泼维的定量下限（LLOQ）为 0.25pg 柱上（进样量为 10μL 时为 25pg/mL）。该方法具有可靠的校准范围，为 0.025-2.5ng/mL。由于在沉淀过程中对实际血浆样品进行了 10 倍稀释，因此该方法适用于定量 0.25-25ng/mL 浓度范围内的博赛泼维和特拉泼维。在整个校准范围内，准确度和日内及日间精密度（每个浓度 6 个样本）的变异均<15%。首次建立并验证了一种快速、特异、灵敏、准确和重现性好的 LC-MS/MS 方法，可用于定量人血浆中 HCV 蛋白酶抑制剂博赛泼维和特拉泼维的浓度。
