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Zorbax Extend-C18柱（150 mm×4.6 mm,5 μm,美国Agilent 公司）；C18保护柱（4.0 mm×3.0 mm,5 μm,迪马公司）；流动相为乙腈-10 mmol/L 醋酸铵-甲酸（60∶40∶0.1,V/V/V）；流速为0.5 ml/min；进样量为20 μl；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/z 315.2～86.1（氯米帕明）和m/z285.1～86.2（内标异丙嗪）.500 μl 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1～500 ng/ml,定量下限为1 ng/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:本方法可用于氯米帕明的犬体内药代动力学研究.%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 mmx4.6 mm, 5 μm, Amercia 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 μl, Mass spectrum condition was electrospray ionization (ESI) and SIM mod 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 all less than 15%. The method recoveries were within 85%-115% and also met for requirement of Chinese Pharmacopoeia (2010) Vol Ⅱ . Conclusion: The method is successfully used for Clomipramine pharmacokinetic studies in dog.
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