UPLC-MS法测定Beagle犬血浆中丁咯地尔和主要代谢物及其在药代动力学研究中的应用  

Determination of buflomedil and its major metabolite in plasma of Beagle dog by UPLC-MS and its application in pharmacokinetic study

作　　者：杨学欣 孙璐[1] 徐静静[1] 宋楠楠 路橦 李乐道[1] 李晨旭 YANG Xuexin, SUN Lu, XU Jingjing, SONG Nannan, LU Tong, LI Ledao, LI Chenxu(School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, Chin)

机构地区：[1]沈阳药科大学药学院,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2018年第3期213-220,共8页Journal of Shenyang Pharmaceutical University

基　　金：辽宁省教育厅项目(L2014390);沈阳药科大学中青年教师中长期培养计划(ZQN2015007);国家大学生创新创业训练计划项目(201710163000217)

摘　　要：目的建立快速灵敏的超高效液相色谱-质谱联用(UPLC-MS)法,研究Beagle犬分别以9.6、19.2、28.8 mg·kg-1的剂量经口给药盐酸丁咯地尔后,丁咯地尔及主要代谢物O-去甲基丁咯地尔在健康Beagle犬体内的药物代谢动力学行为。方法采用甲醇沉淀蛋白法进行血浆样品预处理,Acquity UPLC BEH C18(50 mm×2.1 mm,1.7μm)色谱柱,甲醇-5 mmol·L^(-1)醋酸铵(含体积分数为0.05%的甲酸)为流动相,梯度洗脱。采用ESI源正离子模式对血浆样品中丁咯地尔及其代谢物进行浓度检测,计算药代动力学参数。结果血浆中丁咯地尔和O-去甲基丁咯地尔的质量浓度分别在20.0~4 000.0μg·L^(-1)和2.0~400.0μg·L^(-1)内线性关系良好,日内、日间精密度RSD均不超过10.4%,提取回收率为90.4%~96.3%,基质效应为93.2%~97.0%。丁咯地尔及O-去甲基丁咯地尔血浆样品在所考察的储存条件下均可保持稳定。结论该方法适用于丁咯地尔及O-去甲基丁咯地尔在Beagle犬体内的药物代谢动力学研究。Beagle犬给药9.6~28.8 mg·kg-1剂量内,丁咯地尔药物代谢动力学行为呈线性关系,O-去甲基丁咯地尔药动学行为呈非线性关系。Objective To establish a rapid and sensitive ultra-performance liquid chromatography-mass spectrometry （UPLC-MS） method for the simultaneously determine buflomedil and its major metabolite O- desmethylbuflomedil in dog plasma after oral administration of buflomedil hydrochloride at doses of 9. 6, 19. 2 and 28.8 m· kg^-1 ,respectively, and apply the method to study the pharmacokinetics of buflomedil and O-desmethylbuflomedil in Beagle dogs. Methods The plasma samples were pretreated by protein precipitation with methanol, and chromatographic separation was performed on an Acquity UPLC BEH C18 column using a gradient elution with mobile phase consisting of methanol and 5 mmol·L^-1 NH4AC containing 0. 05 % formic acid. The detection was carded out by Waters XevoTM G2 QTOF equipped with electrospray ionization source in positive ion mode. Results The method was linear over the range of 20. 0 - 4 000. 0 μg·L^-1 for buflomedil and 2.0 - 400.0 μg·L^-1 for O-desmethylbuflomedil, and the intra-day and inter-day precision were below 10. 4%. The extraction recovery were ranged from 90. 4% to 96. 3% ,and the matrix effect were ranged from 93.2% to 97.0%. The plasma samples of the two analytes were found to be stable in the different conditions investigated in this article. Conclusions The method was successfully applied in the pharmacokinetic study of buflomedil and O-desmethylbufiomedil in dog plasma. The pharmacokinetics of buflomedil was linear in the dose range of 9. 6 - 28. 8 mg· kg ^- l in Beagle dogs, while the pharmacokinetic behavior of O-demethylbuflomedil showed a nonlinear relationship during the same dose.

关 键 词：丁咯地尔 O-去甲基丁咯地尔 超高效液相色谱-质谱联用法 药物动力学 

分 类 号：R917[医药卫生—药物分析学]