目的 建立一种快速、灵敏测定Beagle犬血浆中左旋氨氯地平浓度的高效液相色谱-串联质谱(LC-MS/MS)法,研究左旋氨氯地平片在Beagle犬体内的药动学。方法 色谱条件采用Phenomenex Synergi Hydro-RP C₁₈色谱柱(30 mm×2 mm,4 μm);流动相:0.1%甲酸水-0.1%甲酸甲醇,梯度洗脱;体积流量:0.6 mL/min;柱温:室温;进样量:20 μL。质谱条件电喷雾离子源(ESI);多级反应监测(MRM);正离子模式;喷雾气温度(TEM):400 ℃;雾化气(GS1):344.95 kPa;加热辅助气(GS2):413.7 kPa;气帘气(CUR):206.85 kPa;碰撞气(CAD):68.95 kPa;离子喷雾电压(IS):5 500 V;扫描时间:200 ms;用于定量分析的MRM离子对分别为左旋氨氯地平m/z 409.0→238.1,内标硝苯地平m/z 347.0→315.2、347.0→271.3。6只Beagle犬灌服6.8 mg/kg苯磺酸左旋氨氯地平片后,以硝苯地平为内标,血浆样品经醋酸乙酯萃取。绘制左旋氨氯地平的药-时曲线,计算主要药动学参数。结果 左旋氨氯地平在0.05~20.00 ng/mL线性关系良好,定量下限为0.05 ng/mL。批内、批间精密度RSD值均小于10%,平均提取回收率为89.3%~93.6%,基质效应为99.9%~102.7%。主要药动学参数C_max=(6.47±0.72)μg/L,t_max=(2.3±0.5)h,t_1/2=(11.0±4.6)h,MRT=(15.6±6.8)h,AUC_0-t=(68.81±19.29)h·μg/L,AUC_0-∞=(71.58±20.35)h·μg/L。结论 本法特异、灵敏、准确、可靠,可用于Beagle犬血浆中左旋氨氯地平的药物浓度测定及左旋氨氯地平片药动力学研究。

Objective To A rapid and sensitive LC-MS/MS method for determination of levoamlodipine in Beagle dog plasma, and study the pharmacokinetics of Levamlodipin Tablets in Beagle dogs. Methods The analysis was separated on a Phenomenex Synergi Hydro-RP C₁₈ column (250 mm × 4.6 mm, 5 μm), and mobile phase was 0.1% formic acid water-0.1% formic acid methanol by gradient elution at a flow rate of 0.6 mL/min. The column temperature was at 40 ℃, and the injection volume was 20 μL. MS detection was performed with multiple reaction monitoring (MRM) mode using positive electrospray ionization (ESI). TEM was 400 ℃, and GS1, GS2, CUR, and CAD were 344.95, 413.7, 206.85, and 68.95 kPa, respectively. IS was 5 500 V, and scanning time was 200 ms. The ion pairs of m/z 409.0→238.1 and m/z 347.0→315.2 and 347.0→271.3 were used to qualify levamlodipin and internal standard nifedipine. Six Beagle dogs were ig administered with Levamlodipin Tablets 6.8 mg/kg, and nifedipine was used as the internal standard, and the plasma samples were pretreated by ethyl acetate extraction. Mean plasma concentration-time curves of levoamlodipine in Beagle dogs were obtained, and main pharmacokinetic parameters were calculated. Results Levamlodipin has a good linear relation between 0.05—20.0 ng/mL, and the LLOQ was 0.05 ng/mL. RSD values of intra-assay and inter-assay were both less than 10%. The extraction recovery was 89.3%—93.6%. The matrix effect was 99.9%—102.7%. The main pharmacokineticparameters were C_max=(6.47 ± 0.72)μg/L, t_max=(2.3 ± 0.5)h, t_1/2=(11.0 ± 4.6)h, MRT=(15.6 ± 6.8)h, AUC_0-t=(68.81 ± 19.29)h·μg/L, AUC_0-∞=(71.58 ± 20.35)h·μg/L. Conclusion The developed method is specific, sensitive, accuracy and reliable, and it is suitable for determination of levamlodipin in plasma and pharmacokinetic studies of Levamlodipin Tablets in Beagle dogs.

广东省科技计划项目(2015A040404042);广东省产学研专项资金企业创新平台项目(2013B090800013);广东省协同创新与平台环境建设项目(2014B40404066)