Laboratory of Pharmacology and Toxicology, University Hospital, F-42055 Saint-Etienne, France.
J Pharm Biomed Anal. 2012 Jan 25;58:152-6. doi: 10.1016/j.jpba.2011.09.018. Epub 2011 Sep 22.
This article described the development and full validation of rapid and accurate liquid chromatography method, coupled with tandem mass spectrometry detection, for quantification of dabigatran in human plasma with [(13)C(6)]-dabigatran as internal standard. Plasma pretreatment involved a single step protein precipitation with methanol. Separation was performed by ultra performance reversed-phase chromatography on an Acquity UPLC BEH C8 100 mm × 1 mm × 1.7 μm column using a gradient elution mode. The mobile phase was a mix of distilled water containing 0.1% formic acid and methanol containing 0.1% formic acid. Specific multiple reaction monitoring transitions were recorded in positive electrospray ionization. The method was linear over the concentration range of 2-500 μg/L. The intra- and inter-day precision values were below 11.3% and accuracy was within 93.8% and 108.8% for all QC levels (5, 75 and 400 μg/L). The lower limit of quantification was 2 μg/L. Total analysis time was to 10 min including sample preparation.
本文描述了一种快速准确的液相色谱-串联质谱检测法的开发和全面验证，用于定量检测人血浆中的达比加群，内标物为 [(13)C(6)]-达比加群。血浆预处理采用甲醇单步蛋白沉淀法。采用 Acquity UPLC BEH C8 100mm×1mm×1.7μm 柱进行超高效反相色谱分离，采用梯度洗脱模式。流动相为含 0.1%甲酸的蒸馏水和含 0.1%甲酸的甲醇的混合物。在正电喷雾电离模式下记录特定的多重反应监测转换。该方法在 2-500μg/L 的浓度范围内呈线性。所有 QC 浓度（5、75 和 400μg/L）的日内和日间精密度值均低于 11.3%，准确度在 93.8%至 108.8%之间。定量下限为 2μg/L。包括样品制备在内，总分析时间为 10 分钟。
