Objective: To develop a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for simultaneous determination of atorvastatin and voriconazole in rat plasma and investigate the pharmacokinetics of atorvastatin and the changes in voriconazole concentration in rats after administration.
Methods: Plasma samples were collected from rats after intragastric administration of atorvastatin alone or in combination with voriconazole. The samples were treated with sodium acetate acidification, and atorvastatin and voriconazole in the plasma were extracted using a liquidliquid extraction method with methyl tert-butyl ether as the extractant. The extracts were then separated on a Thermo Hypersil Gold C18 (2.1×100 mm, 1.9 μm) column within 6 min with gradient elution using acetonitrile and water (containing 0.1% formic acid) as the mobile phase; mass spectrometry detection was achieved in selective reaction monitoring (SRM) mode under the positive ion scanning mode of heated electrospray ion source (H-ESI) and using transition mass of m/z 559.2→440.2 for atorvastatin and m/z 350→280 for voriconazole, with m/z370.2→252 for lansoprazole (the internal standard) as the quantitative ion.
Results: The calibration curves were linear within the concentration range of 0.01-100 ng/mL (r=0.9957) for atorvastatin and 0.025-100 ng/mL (r=0.9966) for voriconazole. The intra-day and inter-day precisions were all less than 13%, and the recovery was between 66.50% and 82.67%; the stability of the plasma samples met the requirements of testing. The AUC0-24 h of atorvastatin in rat plasma after single and combined administration was 438.78±139.61 and 927.43±204.12 h·μg·L-1, CLz/F was 23.89±8.14 and 10.43±2.58 L·h-1·kg-1, Cmax was 149.62±131.10 and 159.37±36.83 μg/L, t1/2 was 5.08±1.63 and (5.58±2.11 h, and Tmax was 0.37±0.14 and 3.60±1.52 h, respectively; AUC0-24 h, CLZ/F and Tmax of atorvastatin in rat plasma differed significantly between single and combined administration. The HPLC-MS/MS system also allowed simultaneous determination of voriconazole concentration in rat plasma after combined administration.
Conclusions: The HPLC-MS/MS system we established in this study is simple, rapid and sensitive and allows simultaneous determination of atorvastatin and voriconazole in rat plasma. Some pharmacokinetic parameters of atorvastatin are changed in the presence of voriconazole, and their clinical significance needs further investigation.
目的: 建立并应用同时测定大鼠血浆中阿托伐他汀和伏立康唑浓度的液相质谱联用方法(HPLC-MS/MS), 研究阿托伐他汀单独给药及与伏立康唑联合给药后, 大鼠体内阿托伐他汀的药动学变化以及伏立康唑的浓度变化。
方法: 采用醋酸钠酸化, 甲基叔丁基醚液-液萃取法处理血浆样品, 经Thermo Hypersil Gold C18 (2.1×100 mm, 1.9 μm)色谱柱分离, 流动相为乙腈-0.1%甲酸水溶液, 梯度洗脱, 分析时间6 min; 质谱检测采用加热电喷雾离子源(H-ESI)正离子扫描方式, 选择反应监测(SRM)模式检测, 选用m/z559.2→440.2 (阿托伐他汀)、m/z350.2→281 (伏立康唑)、m/z370.2→252 (内标兰索拉唑)作为定量分析的离子。
结果: 阿托伐他汀在0.01~100 ng/mL (r=0.9957), 伏立康唑在0.025~100 ng/mL (r=0.9966)范围内线性关系良好, 阿托伐他汀和伏立康唑的日间和日内精密度均小于13%, 提取回收率66.50%~82.67%, 血浆样品稳定性符合测定要求。单独给药和联合给药后大鼠血浆中阿托伐他汀的AUC0-24 h分别为(438.78±139.61)和(927.43±204.12))h·μg·L-1, CLz/F分别为(23.89±8.14)和(10.43± 2.58)L·h-1·kg-1, Cmax分别为(149.62±131.10)和(159.37±36.83)μg·L-1, t1/2分别为(5.08±1.63)和(5.58±2.11)h, Tmax分别为(0.37± 0.14)和(3.60±1.52)h, 其中AUC0-24 h和CLz/F, Tmax具有显著性差异(P < 0.05)。同时可测定合并给药组中伏立康唑的血药浓度。
结论: 本方法简单快速, 灵敏度高, 可用于同时检测大鼠血浆中阿托伐他汀和伏立康唑的浓度。伏立康唑和阿托伐他汀联用, 使阿托伐他汀的部分药代动力学参数发生了改变。
Keywords: atorvastatin; high-performance liquid chromatography-tandem mass spectrometry; interaction; pharmacokinetics; voriconazole.