Barbara Ann Karmanos Cancer Institute, Wayne State University, Detroit, MI 48201, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Feb 1;878(3-4):333-9. doi: 10.1016/j.jchromb.2009.11.037. Epub 2009 Nov 27.
A reversed-phase liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) method was developed and validated for simultaneous determination of ABT-888 and its major metabolite (M8) in human plasma. Sample preparation involved a liquid-liquid extraction by the addition of 0.25 ml of plasma with 10 microl of 1 M NaOH and 1.0 ml ethyl acetate containing 50 ng/ml of the internal standard zileuton. The analytes were separated on a Waters XBridge C(18) column using a gradient mobile phase consisting of methanol/water containing 0.45% formic acid at the flow rate of 0.2 ml/min. The analytes were monitored by tandem mass spectrometry with electrospray positive ionization. Linear calibration curves were generated over the ABT-888 and M8 concentration ranges of 1-2000 ng/ml in human plasma. The lower limits of quantitation (LLOQ) were 1 ng/ml for both ABT-888 and M8 in human plasma. The accuracy and within- and between-day precisions were within the generally accepted criteria for bioanalytical method (<15%). This method was successfully employed to characterize the plasma concentration-time profile of ABT-888 after its oral administration in cancer patients.
建立并验证了一种反相高效液相色谱-串联质谱（LC-MS/MS）法，用于同时测定人血浆中的 ABT-888 及其主要代谢物（M8）。样品制备采用液液萃取法，加入 0.25ml 血浆，用 10μl 1M NaOH 和 1.0ml 含 50ng/ml 内标的齐留通的乙酸乙酯进行提取。采用 Waters XBridge C18 柱，以甲醇/水（含 0.45%甲酸）为流动相，流速为 0.2ml/min，进行梯度洗脱分离。采用电喷雾正离子化串联质谱法进行检测。ABT-888 和 M8 在人血浆中的浓度范围为 1-2000ng/ml 时，线性校准曲线均良好。ABT-888 和 M8 在人血浆中的定量下限（LLOQ）均为 1ng/ml。准确度和日内、日间精密度均符合生物分析方法的一般接受标准（<15%）。该方法成功用于测定癌症患者口服 ABT-888 后的血浆浓度-时间曲线。
