目的 建立一種快速、靈敏測(cè)定Beagle犬血漿中左旋氨氯地平濃度的高效液相色譜-串聯(lián)質(zhì)譜(LC-MS/MS)法,研究左旋氨氯地平片在Beagle犬體內(nèi)的藥動(dòng)學(xué)。方法 色譜條件采用Phenomenex Synergi Hydro-RP C₁₈色譜柱(30 mm×2 mm,4 μm);流動(dòng)相:0.1%甲酸水-0.1%甲酸甲醇,梯度洗脫;體積流量:0.6 mL/min;柱溫:室溫;進(jìn)樣量:20 μL。質(zhì)譜條件電噴霧離子源(ESI);多級(jí)反應(yīng)監(jiān)測(cè)(MRM);正離子模式;噴霧氣溫度(TEM):400 ℃;霧化氣(GS1):344.95 kPa;加熱輔助氣(GS2):413.7 kPa;氣簾氣(CUR):206.85 kPa;碰撞氣(CAD):68.95 kPa;離子噴霧電壓(IS):5 500 V;掃描時(shí)間:200 ms;用于定量分析的MRM離子對(duì)分別為左旋氨氯地平m/z 409.0→238.1,內(nèi)標(biāo)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,以硝苯地平為內(nèi)標(biāo),血漿樣品經(jīng)醋酸乙酯萃取。繪制左旋氨氯地平的藥-時(shí)曲線,計(jì)算主要藥動(dòng)學(xué)參數(shù)。結(jié)果 左旋氨氯地平在0.05~20.00 ng/mL線性關(guān)系良好,定量下限為0.05 ng/mL。批內(nèi)、批間精密度RSD值均小于10%,平均提取回收率為89.3%~93.6%,基質(zhì)效應(yīng)為99.9%~102.7%。主要藥動(dòng)學(xué)參數(shù)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。結(jié)論 本法特異、靈敏、準(zhǔn)確、可靠,可用于Beagle犬血漿中左旋氨氯地平的藥物濃度測(cè)定及左旋氨氯地平片藥動(dòng)力學(xué)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.

廣東省科技計(jì)劃項(xiàng)目(2015A040404042);廣東省產(chǎn)學(xué)研專項(xiàng)資金企業(yè)創(chuàng)新平臺(tái)項(xiàng)目(2013B090800013);廣東省協(xié)同創(chuàng)新與平臺(tái)環(huán)境建設(shè)項(xiàng)目(2014B40404066)