Department of Pharmacokinetics and Drug Metabolism, Amgen, Thousand Oaks, CA 91320-1799, United States.
J Pharm Biomed Anal. 2011 Nov 1;56(3):604-8. doi: 10.1016/j.jpba.2011.06.022. Epub 2011 Jul 1.
The goal of this work was to obtain full pharmacokinetic profiles from individual mice with the use of an automated blood sampling system and dried blood spot (DBS) technique. AMG 517, a potent and selective vanilloid receptor (VR1) antagonist, was dosed to mice (n=3) intravenously and blood samples were collected using the automated blood sampling system with the "no blood waste" method. The collected blood samples were a mixture of 25 μL blood and 50 μL of heparinized saline solution. Two 15 μL aliquots were manually spotted onto a DBS card and dried at room temperature for at least 2h before being stored in zip bags with desiccant. The remaining samples (45 μL) were stored at -70°C until analysis. Both the DBS and the whole blood samples (diluted with saline (1:2, v/v)) were extracted and analyzed by liquid chromatography-tandem mass spectrometry. The overall extraction recovery of the analyte from the dried blood spots was determined to be about 90%. The pharmacokinetic parameters calculated using the whole blood or the DBS concentration data were comparable, and were obtained from only 3 mice, whereas conventional sampling and analysis would have required up to 27 mice to achieve the same result. The analyte was shown to be stable in the diluted whole blood (blood:saline 1:2) at room temperature for at least 4h and in the DBS for at least 34 days when stored at room temperature. These results indicated that the automated blood sampling system and DBS collection are promising techniques to obtain full pharmacokinetic profiles from individual mice and reduce the use of animals.
本研究旨在利用自动化采血系统和干血斑（DBS）技术从个体小鼠中获得完整的药代动力学曲线。AMG 517 是一种强效且选择性的香草素受体（VR1）拮抗剂，对小鼠进行静脉注射给药，并采用“无血液浪费”的自动化采血系统采集血样。采集的血样是 25μL 血液与 50μL 肝素化生理盐水的混合物。手动吸取 2 个 15μL 等分试样点样至 DBS 卡上，在室温下干燥至少 2h 后，将其储存在装有干燥剂的拉链袋中。其余样品（45μL）在-70°C 下储存，直至分析。通过液相色谱-串联质谱法对 DBS 和全血样品（用生理盐水（1:2，v/v）稀释）进行提取和分析。从干血斑中提取分析物的总体提取回收率约为 90%。使用全血或 DBS 浓度数据计算得到的药代动力学参数具有可比性，并且仅使用 3 只小鼠即可获得，而常规采样和分析则需要多达 27 只小鼠才能获得相同的结果。结果表明，在室温下，稀释后的全血（血液：生理盐水 1:2）中至少 4h 以及 DBS 中至少 34 天内，分析物均稳定。这些结果表明，自动化采血系统和 DBS 采集是从个体小鼠中获得完整药代动力学曲线并减少动物使用的有前途的技术。
