目的:建立LC-M S/M S测定人血浆中肌苷浓度的方法并应用于异丙肌苷药代动力学研究。方法以阿德福韦为内标,采用甲醇∶10 mmol/L 乙酸铵(15∶85,v/v )为流动相,以Agilent SB-C18柱(5μm粒径,150.0 mm ×4.6 mm I .D .)为分析柱,通过电喷雾电离源(ESI),M RM扫描方式进行检测。用于定量分析的离子反应分别为m/z母离子为267.3,子离子为135.0(肌苷)和m/z母离子为272.0,子离子为134.1(阿德福韦)。结果血浆中肌苷定量的线性范围10~3000 ng/m L ,定量下限为10 ng/mL。日内、日间精密度(RSD)小于5%,平均回收率大于90%,无基质效应。结论本法专属性强,样品处理方便,灵敏度高,适用于肌苷临床药动学研究。
A rapid and sensitive method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the determination of a novel anticoagulant peptide bivalirudin in human plasma has been developed and validated. Plasma samples were precipitated protein with acetonitrile and re-extracted with dichloromethane, after which the analyte and triptorelin as an internal standard (IS) were separated on a 300SB-C18 column (150 mm 4.6 mm i.d., 5 mm particle size) using 0.1% formic acid:methanol (45:55, v/v) as mobile phase. The triple-quadrupole mass spectrometer, equipped with electrospray ionization (ESI) interface, was operated in the positive ion mode, and the multiple-reaction monitoring (MRM) transitions of bivalirudin and IS were at m/z 1091.0-650.4 and m/z656.5-249.3, respectively. The lower limit of quantification (LLOQ) was 1 ng/mL for 100 mL plasma sample and the assay was linear over the concentration range 1-1000 ng/mL. The accuracy was within a range from 0.4% to 0.5% in terms of relative error (RE) and the intra- and inter-day precisions in terms of relative standard deviation (RSD) were r2.92 and r3.36, respectively. The method was successfully applied to a pharmacokinetic study involving intravenous administration of bivalirudin (0.5 mg/kg) to Chinese volunteers.