中国临床药学杂志

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(50 mm×2.1 mm,1.8μm),以乙腈-水(含0.1%甲酸)为流动相进行梯度洗脱,流速为0.4 m L·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(Necrostatin-1)和285.1→192.9(内标:地西泮)。结果 Necrostatin-1的线性范围为4~2500μg·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6 h、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Necrostatin-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 t1/2 was 1. 89 h. CONCLUSION The established method is proved to be suitable for determination of Necrostatin-1 and its pharmacokinetic study in rat plasma.