Zhou Ying, Li Huqun, He Xiaomeng, Jia Mengmeng, Ni Yang, Xu Mingzhen, Chen Hui, Li Weiyong
Institute of Clinic Pharmacy, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Institute of Clinic Pharmacy, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
J Pharm Biomed Anal. 2014 Nov;100:294-299. doi: 10.1016/j.jpba.2014.08.018. Epub 2014 Aug 17.
A simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of clevidipine and its primary metabolite H152/81 in dog plasma after protein precipitation with acetonitrile using felodipine as the internal standard (IS). Chromatographic separation was performed on a XB C18 column (2.1mm×50mm, 3.5μm) under isocratic conditions with the mobile phase consisting of acetonitrile and 20mM ammonium acetate buffer (pH 7.0) (40:60, v/v) at the flow rate of 0.3ml/min. The run time was 5.5min. Mass spectrometric analysis was performed on a triple quadrupole mass spectrometer operated in the multiple reaction monitoring (MRM) mode with the transitions of m/z 473.0→338.2 for clevidipine, m/z 356.1→324.0 for H152/81 and m/z 383.9→338.2 for the IS. The method was fully validated in terms of selectivity, linearity, lower limit of quantification (LLOQ), accuracy, precision, stability, matrix effect and recovery over a concentration range of 0.15-200ng/ml for clevidipine and 10-2000ng/ml for H152/81, respectively. The analytical method was applied to support a pharmacokinetic study of simultaneous determination of clevidipine and H152/81 in ten healthy beagle dogs.
建立了一种简单快速的液相色谱-串联质谱（LC-MS/MS）方法，并进行了验证，用于在以非洛地平为内标（IS）用乙腈进行蛋白沉淀后，同时测定犬血浆中的氯维地平及其主要代谢物H152/81。在XB C18柱（2.1mm×50mm，3.5μm）上于等度条件下进行色谱分离，流动相由乙腈和20mM醋酸铵缓冲液（pH 7.0）（40:60，v/v）组成，流速为0.3ml/min。运行时间为5.5分钟。在三重四极杆质谱仪上以多反应监测（MRM）模式进行质谱分析，氯维地平的质荷比转换为m/z 473.0→338.2，H152/81为m/z 356.1→324.0，内标为m/z 383.9→338.2。该方法在选择性、线性、定量下限（LLOQ）、准确度、精密度、稳定性、基质效应和回收率方面进行了全面验证，氯维地平的浓度范围为0.15 - 200ng/ml，H152/81的浓度范围为10 - 2000ng/ml。该分析方法用于支持一项在十只健康比格犬中同时测定氯维地平和H152/81的药代动力学研究。
