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目的 采用LC-MS/MS法测定大鼠血浆中Necrostatin-1的浓度,并应用于大鼠体内药动学研究.方法 血浆样品经乙腈蛋白沉淀法处理后进行分析.采用SB-C18色谱柱(50mm×2.1 mm,1.8 μm),以乙腈-水(含0.1％甲酸)为流动相进行梯度洗脱,流速为0.4 mL·min-1,正离子多离子反应监测(MRM)检测.用于定量分析的离子对为m/z 260.1→131.0(Ne-crostatin-1)和285.1→192.9(内标:地西泮).结果 Necrostatin-1的线性范围为4～ 2500 μg· L-1,最低定量下限为4μg·L-1;提取回收率为85.49％～93.21％,基质效应为93.25％～101.11％;日内、日间RSD均＜10％,血浆样品室温放置6h、3次冻融循环以及低温(-70℃)储存2周稳定.Necrostatin-1的药动学参数t1/2为1.89 h.结论 本方法学符合生物样品分析要求,可用于大鼠血浆N ecrostatin-1药物浓度的测定及其药动学研究.“,”AIM To develop an LC-MS/MS method for determination of Necrostatin-1 concentration in rat plasma,and further apply the method in pharmacokinetic study.METHODS Specimens were treated with acetonitrile protein precipitation method and then separated on an SB-C18 (50 mm × 2.1 mm,1.8 μm) column with the mobile phase of acetonitrile/water (0.1％ formic acid) at a flow rate of 0.4 mL · min-1.The mass spectrometerwas operated in multiple reaction monitoring(MRM) mode with positive electrospray ionization.The precursor ion→product ion transitions m/z 260.1→131.0 for Necrostatin-1 and m/z 285.1→192.9 for diazepam(internal standard) were monitored.RESULTS The plasma standard curve was linear in the range of 4-2 500 μg · L-1.The limit of quantification was 4 μg · L-1.The extraction recoveries were within the range of 85.49％-93.21％.Matrix effect was 93.25％-101.11％.The intra-day and inter-day precision relative standard deviations (RSD) were less than 10％.Samples were placed at room temperature for 6 h,3 times of freeze-thaw cycles and low temperature (-70℃) for 2 weeks for stabilization.The t1/2was 1.89 h.CONCLUSION The established method is proved to be suitable for determination of Necrostatin-1 and its pharmacokinetic study in rat plasma.