超高效液相色谱-串联质谱法同时测定食蟹猴血浆中地高辛、安非他酮及活性代谢物浓度和药动学评价  

Simultaneous determination of digoxin, bupropion and its active metabolites in cynomolgus monkey plasma by UPLC-MS/MS and its application in pharmacokinetic study

作　　者：于洋[1,2] 何佳珂 夏春华[2] YU Yang;HE Jia-ke;XIA Chun-hua(Department of Pharmacy, The Second Affiliated Hospital of Nanchang University, Nanchang Jiangxi 330006, China;Institute of Clinical pharmacology,Department of Pharmacy, Nanchang University, Nanchang Jiangxi 330006, China)

机构地区：[1]南昌大学第二附属医院药学部,江西南昌330006 [2]南昌大学临床药理研究所,江西南昌330006

出　　处：《中国医院药学杂志》2019年第14期1460-1466,共7页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金(81603188);中国博士后科学基金面上项目(2017M612165);江西省博士后科研项目(2017KY24);江西省自然科学基金青年项目(20181BAB215045);江西省教育厅科学技术研究项目青年项目(GJJ170156)

摘　　要：目的:建立同时测定食蟹猴体内地高辛、安非他酮及其活性代谢物血浆浓度的方法,并评价药动学特征。方法:以乙腈沉淀蛋白处理血浆样品。色谱柱Agilent Zorbax SB-C8(4.6×100 mm 3.5μm);流动相为甲醇-水(含5 mmol·L-1甲酸铵,pH 3.6),梯度洗脱;质谱条件为电喷雾离子源,正离子模式,扫描方式为多反应离子监测,m/z 798.4→651.3 (地高辛);m/z 240.2→184.0 (安非他酮);m/z 256.3→238.0 (羟基安非他酮);m/z 242.2→168.1 (赤式/苏式羟化安非他酮);m/z 172.2→128.2 (甲硝唑,内标)。食蟹猴3只,♂,体质量2~4 kg,0.1 mg·5 mL-1·kg-1地高辛和1.5 mg·5 mL-1·kg-1盐酸安非他酮静滴给药。给药前后0.08,0.25,0.5,1,1.5,2,4,6,8,12,24,48 h采血。测定地高辛、安非他酮及其活性代谢物浓度。结果:血浆标准曲线在0.25~100 ng·mL-1(地高辛),0.2~1 000 ng·mL-1(安非他酮/羟基安非他酮),0.2~50 ng·mL-1(赤式/苏式羟化安非他酮),定量范围内线性良好(r≥0.995)。批内批间精密度均小于13.7%,绝对回收率为83.8%~104.2%,基质效应小于10.6%。地高辛、安非他酮、羟基安非他酮、赤式、苏式羟化安非他酮药动学参数Cmax分别为(29.32±7.31)、(254.00±68.23)、(22.43±7.56)、(1.53±0.27)、(3.96±0.42) ng·mL-1,Tmax分别为(0.14±0.10)、(0.08±0)、(1.50±0.00)、(1.17±0.29)、(4.06±1.46) h,AUC0-48h分别为(123.87±9.59)、(550.68±17.43)、(160.47±62.92)、(9.26±3.65)、(24.43±5.28) ng·h·mL-1,半衰期分别为(21.26±3.77)、(5.54±1.76)、(3.96±1.21)、(5.58±2.40)、(4.06±1.46) h。结论:和已报道的方法比较,建立的分析方法灵敏、准确,样品处理简便、快速,适用于药动学研究以及地高辛-安非他酮药物相互作用的评价,也能为临床个体化用药实践提供方法学参考。OBJECTIVE To establish a method for simultaneous determination of digoxin(DIG), bupropion(BUP) and their active metabolites in cynomolgus monkey plasma and evaluate their pharmacokinetic characteristics. METHODS The plasma samples were treated with protein precipitation with acetonitrile. Chromatographic column Agilent Zorbax SB-C8(4.6×100 mm 3.5 μm);mobile phase: methanol-water(containing 5 mM ammonium formate, pH 3.6), gradient elution;mass spectrometry conditions: electrospray ionization source, positive mode;scanning mode: multiple reaction ion monitoring,: m/z 798.4→651.3(DIG), m/z 240.2→184.0(BUP), m/z 256.3→238.0(HBUP), m/z 242.2→168.1(EBUP/TBUP), m/z 172.2→128.2(metronidazole, internal standard). 3 male cynomolgus monkeys with a weight of 2-4 kg were intravenously administered at 0.1 mg·(5 mL)^-1·kg^-1 DIG and 1.5 mg·(5 mL)^-1·kg^-1 BUP. Blood samples were collected at 0.08, 0.25, 0.5, 1, 1.5, 2, 4, 6, 8, 12, 24, and 48 h before and after administration. The concentrations of digoxin, bupropion and their active metabolites were determined. RESULTS The plasma standard curve showed good linearity(r≥0.995) in the range of 0.25-100 ng·mL-1(digoxin), 0.2-1 000 ng·mL^-1(bupropion/hydroxybupropion), and 0.2-50 ng·mL-1(erythro-/threohydroxybupropion). Intra-assay inter-assay precision was less than 13.7%, absolute recovery was 83.8-104.2%, and matrix effect was less than 10.6%. Cmax were(29.32±7.31),(254.00±68.23),(22.43±7.56),(1.53±0.27),(3.96±0.42) ng·mL^-1, Tmax were(0.14±0.10),(0.08±0),(1.50±0.00),(1.17±0.29),(4.06±1.46) h, AUC0-48 h were(123.87±9.59),(550.68±17.43),(160.47±62.92),(9.26±3.65),(24.43±5.28) ng·h·mL^-1, t1/2 were(21.26±3.77),(5.54±1.76),(3.96±1.21),(5.58±2.40),(4.06±1.46) h for DIG, BUP, HBUP, EBUP and TBUP, respectively.CONCLUSION Compared with the reported method, the established method is sensitive, accurate, simple and rapid, and suitable for pharmacokinetic study and evaluation of drug interaction of digoxin and bupropion.

关 键 词：地高辛 安非他酮 安非他酮代谢物 超高效液相色谱-串联质谱法 药动学 

分 类 号：R917[医药卫生—药物分析学]