LC-MS/MS法同时测定Beagle犬血浆中阿托伐他汀钙和盐酸吡格列酮的浓度  

Concentration determination of atorvastatin calcium and pioglitazone hydrochloride in Beagle dog plasma by LC-MS/MS

作　　者：陈和莉[1,2] 张文萍[2,3] 杨付英[2] 魏世杰[2,3] 党宏万[2,3] 

机构地区：[1]第三军医大学西南医院药学部,重庆400038 [2]宁夏医科大学药学院,银川750004 [3]宁夏医科大学总医院药剂科临床药理研究室,银川750004

出　　处：《药物分析杂志》2013年第8期1311-1316,1321,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：宁夏自然科学基金项目(NZ1205)

摘　　要：目的:建立LC-MS/MS方法测定Beagle犬血浆中阿托伐他汀钙和盐酸吡格列酮的浓度,并验证方法。方法:采用蛋白沉淀法提取血浆中药物,以地西泮为内标;使用Shim-pack XR-ODS(2.0 mm×100 mm,2.2μm)色谱柱,Shim-pack GVP-ODS(2.0 mm×5 mm,2.2μm)保护柱,柱温40℃;流动相A为5 mmol.L-1醋酸铵(pH 5.55),B为乙腈,采用梯度洗脱(0～0.6 min,10%B;0.6～1.2 min,10%→90%B;1.2～3.5 min,90%B;3.5～4.1 min,90%→10%B,5.5 min结束);流速为0.3 mL.min-1;进样体积20μL。质谱均采用电喷雾离子源(ESI),以多反应离子监测(MRM)模式进行定量分析。结果:阿托伐他汀钙、盐酸吡格列酮分别在0.05～50、0.5～1000 ng.mL-1范围内有良好的线性关系,最低检测限分别为0.05、0.5 ng.mL-1,日内、日间精密度RSD均<15%(n=5)。含药血浆室温放置24 h、处理后4℃放置24 h、反复冻融3次及-70℃下放置2个月稳定。结论:本方法简便、准确,能用于Beagle犬血浆中阿托伐他汀钙、盐酸吡格列酮的药物浓度测定和药动学研究。Objective： To establish an LC-MS /MS method to determine the concentrations of atorvastatin calcium and pioglitazone hydrochloride in Beagle dog plasma,and to verify the method.Methods： The drugs were extracted by precipitating protein,with diazepam as an internal standard.The Shim-pack XR-ODS（2.0 mm × 100 mm,2.2 μm） column at 40 ℃ and the Shim-pack GVP-ODS（2.0 mm ×5 mm,2.2 μm） pre-column were used.The mobile phase consisted of phase A（5 mmol.L-1 ammonium acetate,pH 5.55） and phase B（acetonitrile）,the phase B accounted for 10% of the total mobile phase ratein the first 0.6 min,then changed to 90% from 0.6 min to 1.2 min,and kept at 90% until 3.5 min,then returned to 10% from 3.5 min to 4.1 min.The analysis was stopped until 5.5 min,the total mobile phase rate was 0.3 mL.min-1,injected volume was 20 μL.Electrospray ion source was applied in mass spectrometry,and quantitative analysis was undertaken in multiple reaction monitoring.Results： Atorvastatin calcium and pioglitazone hydrochloride had a good linear relation between 0.05-50 ng.mL-1,0.5 1000 ng.mL-1,respectively.The intra-and inter-day precision RSD were both less than 15%（n =5）.Drugs in dog plasma were stable at room temperature for 24 h,and at 4 ℃ for 24 h after being extracted,even after freezing and thawing three times and stored at-70 ℃ for two months.Conclusion： The developed method is simple and accurate,which can be used to determine the concentration and the pharmacokinetics of atorvastatin calcium and pioglitazone hydrochloride in Beagle dogs.

关 键 词：阿托伐他汀钙 盐酸吡格列酮 LC-MS MS BEAGLE犬血浆 药物浓度测定 药代动力学 

分 类 号：R917[医药卫生—药物分析学]