高效液相色谱-串联质谱法测定犬血浆中的普瑞巴林及在药代动力学中的应用  

High performance liquid chromatography tandem mass spectrometry method for the quantification of pregabalin in dog plasma and its application in pharmacokinetics study

作　　者：王瑜[1] 仲云熙[1] 金孝亮 谷世寅 彭英[1] 尹莉芳[2] 贾元威[3] 孙建国[1] 王广基[1] 

机构地区：[1]中国药科大学药物代谢动力学重点实验室 [2]中国药科大学药物制剂实验室 [3]皖南医学院第一附属医院弋矶山医院临床药学部

出　　处：《中国临床药理学与治疗学》2016年第7期731-738,共8页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：十三五重大专项(2014ZX0950Y004-002);安徽省教育厅2016年高校优秀青年人才支持计划重点项目(gxyq ZD2016178;gxyq ZD2016179)

摘　　要：目的：建立并验证一种测定犬血浆中普瑞巴林浓度的高效、灵敏、快速的液相色谱串联质谱（LC-MS/MS）检测方法,并应用于Beagle犬单次口服给予普瑞巴林胶囊后体内的药代动力学研究。方法：犬血浆样品经甲醇沉淀蛋白后,使用Agilent ZORBAX-C18（2.1 mm×150 mm,3.5μm）色谱柱,以含0.1%甲酸的水和甲醇作为流动相进行梯度洗脱。采用电喷雾离子源（ESI）正离子模式下多反应监测模式（MRM）分析测定,普瑞巴林及非那西汀的离子对分别为[M＋H]＋m/z 160.4→83.3和[M＋H]＋m/z 180.3→110.2。6只Beagle犬口服给予150 mg普瑞巴林胶囊后采集不同时间点血样,用建立的LC-MS/MS分析方法进行测定。结果：普瑞巴林在30~10 000 ng/m L范围内线性良好（r=0.999 5）,提取回收率为101.2%~102.9%,批内与批间精密度符合生物样品的分析要求。Beagle犬口服给予150 mg普瑞巴林胶囊后,Cmax为（25 637.5±2 571.8）ng/m L,Tmax为（1.0±0.5）h,t1/2为（4.4±0.6）h,MRT为（6.7±0.7）h。结论：此方法成功地应用于单次口服给予Beagle犬150 mg普瑞巴林胶囊的药代动力学研究。ABSTRACT AIM： To develop and validate an efficient, rapid and specific high-performance liquid chromatography tandem mass spectrometry （LC-MS/ MS） method for quantification of pregabalin in bea- gle dog plasma and apply to pharmacokinetics study in Beagle dogs. METHODS： After protein precipi- tation, pregabalin and phenacetin （internal stand- ard） were separated using an Agilent ZORBAX-C18 column （2.1 mm×150 mm, 3.5-Micron） with gra- dient elution. The analytes were detected by an elec- trospray positive ionization mass spectrometry in the multiple reaction monitoring mode. The transition （m/z 160.4→83.3） and （m/z 180.3→110.2） were used for quantification of pregabalin and phe- nacetin respectively. Six Beagle dogs were orally ad- ministered with pregabalin capsule （ 150 mg/dog） and serial blood samples were collected at different times and analyzed with this LC-MS/MS method. RESULTS： The calibration curves of pregabalin showed a good linearity over the concentration range of 30 - 10 000 ng/mL. The average recoveries of pregabalin in dog plasma ranged from 101.2% to 102.9%. The results of intra- and inter-batch accuracy and precision met the requirements. After sin- gle oral administration of pregabalin capsule （150 rag/dog）, the Cmax was （25 637.5 ±2 571.8） ng/mL and Tmax was （1.0±0.5） h. The mean plasma elimination half-life and mean residence time were （4.4±0.6） h and （6.7±0.7 ） h respectively. CONCLUSION： The method was successfully applied to the pharmacokinetics study of pregabalin capsule （150 rag） administered as a single oral dose in dogs.

关 键 词：普瑞巴林 LC-MS/MS 药物代谢动力学 BEAGLE犬 

分 类 号：R969[医药卫生—药理学]