LC／MS／MS测定犬血浆中氯米帕明浓度及体内药动学

目的：建立液相色谱-串联质谱法测定犬血浆中氯米帕明（Clomipramine）。方法：色谱柱为ZorbaxExtend—C18柱（150mm×4．6mm，5μm，美国Agilent公司）；C18保护柱（4．0mm×3．0mm，5μm，迪马公司）；流动相为乙腈-10mmol／L醋酸铵-甲酸（60：40：0．1，V／V／V）；流速为0．5ml／min；进样量为20μl；柱温为20℃；采用三重四极杆串联质谱仪，ESI电离源，以选择反应监测方式进行检测。用于定量分析的离子反应分别为m／z315．2，86．1（氯米帕明）和m／z285．1～86．2（内标异丙嗪）。500μl血浆样品经正己烷液提取后蒸干，流动相复溶，进行分析。结果：测定血浆中氯米帕明的线性范围为1～500ng／ml，定量下限为1ng／ml；日内、日间精密度RSD均小于15％，准确度在85％。115％范围内．符合《中国药典》2010年版二部的规定．结论：本方法可用干氯米帕明的女体内药代动力学研究。

Study on the concentration and pharmacokinetic of Clomipramine in dog plasma by LC/MS/MS

Objective： To establish an LC/MS/MS method for the determination of Clomipramine in dog plasma. Methods： The determination was carried on Zorbax Extend-C18 （150 mm×4.6 mm, 5 μm, Amereia agilent） with the mobile phase consisted of acetonitrile-10 mmol/L ammonium acetate solution-formic acid （60：40：0.1, V/V/V）. Column temperature was set at 20℃, flow rate of mobile phase was 0.5 ml/min, the sample amount was 20 Ixl. Mass speetrum condition was electrospray ionization （ESI） and SIM rood selected ion mass spectral m/z 315.2-86.1 （Clomipramine） and m/z 285.1-86.2 （internal standard Promethazine）. The plasma samples with 500 μl which were extracted with N-hexane and reconstituted with mobile phase were analysed. Results： The calibration curve was established in the concentration range of 1-500 ng/ml. The lowest limit of quantification was 1 ng/ml. RSD of inter-day and intra-day precision were 811 less than 15%. The method recoveries were within 85%-115% and also met for requirement of Chinese Pharmacopoeia （2010） Vol 11 . Conclusion： The method is suecessfully used for Clomipramine pharmacokinetic studies in dog.