LC-MS/MS法测定犬血浆中华法林钠的浓度及其应用

Determination of warfarin sodium in canine plasma by LC-MS / MS and its pharmacokinetic study

目的建立LC-MS/MS联用的方法测定犬血浆中华法林钠的浓度并进行药动学研究。方法以卤米松作为内标,犬血浆经甲醇蛋白沉淀处理后稀释进样。色谱柱为Venusil XBP苯基柱(100 mm×2.1 mm,5μm),流动相为0.1%甲酸水溶液(含2 mmol·L-1醋酸铵)-甲醇(含0.1%甲酸)(25∶75),流速0.3 m L·min-1;进样体积5μL。采用电喷雾离子源,负离子方式多反应监测(MRM)扫描分析。检测离子通道分别为m/z 307.1→161.3(华法林钠)和m/z 489.5→413.1(卤米松)。结果华法林钠的线性范围为5~1 500μg·L-1(r=0.997 8),批内和批间精密度RSD均<8%,高、中、低浓度的方法回收率为103.33%~107.29%。结论本方法操作简捷,灵敏度高,专属性好,适用于华法林钠的药动学研究。

AIM To establish an LC-MS /MS method for the determination of warfarin sodium in the canine plasma. METHODS After the addition of halometasone( internal standards,IS),the canine plasma was achieved by protein precipitation using methanol. The analyses were performed using a Venusil XBP Phenyl column( 100 mm × 2. 1 mm,5 μm) at a flow rate of 0. 3 m L·min-1. The mobile phase consisted of 1% formic acid water solution( containing 2 mmol·L-1ammonium acetate)-1% methyl alcohol methanoic acid( 25∶ 75),using isocratic elution,and injection volume was 5 μL. The pronated ions of the analytes were detected in negative ionization in multiple reaction monitoring( MRM) mode. The mass transition pairs of m / z 307. 1 →161. 3 and 489. 5 →413. 1were used to detect warfarin sodium and halometasone,respectively. RESULTS The calibration curve was linear over the concentration range of 5 to 1 500 μg·L-1( r = 0. 997 8). The intra-and inter-day relative standard deviations were less than 8 %. The recovery ranged from 103. 33 % to 107. 29 % at high,middle and low concentrations,respectively. CONCLUSION The method is simple,accurate and sensitive,and suitable for the determination of warfarin sodium.