UPLC-MS/MS法同时测定大鼠血浆中安非他酮及其活性代谢产物的浓度  

Simultaneous Determination of Bupropion and Its Metabolites in Rat Plasma by UPLC-MS/MS

作　　者：于洋[1,2] 何佳珂 熊爱珍[1] 夏春华[2] YU Yang;HE Jia-ke;XIONG Ai-zhen;XIA Chun-hua(Department of Pharmacy of the Second Affiliated Hospital Nanchang 330006,China;Institute of Clinical Pharmacology,Nanchang University,Nanchang 330006,China)

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

出　　处：《南昌大学学报（医学版）》2019年第1期1-6,12,共7页Journal of Nanchang University：Medical Sciences

基　　金：国家自然科学基金项目(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(2018BAB215045);江西省教育厅科学技术研究项目青年项目(GJJ170156)

摘　　要：目的建立同时测定大鼠体内安非他酮及其活性代谢产物血浆浓度的方法,并应用于药动学研究。方法采用超高效液相色谱-串联质谱法(UPLC-MS/MS)。色谱柱为Agilent Zorbax SB-C8 4.6 mm×100 mm,3.5μm;流动相为水-甲醇(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,离子对分为m/z240→m/z184(安非他酮)、m/z256→m/z238(羟基安非他酮)、m/z242→m/z168(赤式/苏式羟化安非他酮)、m/z285→m/z154(地西泮,内标)。取3只雌性SD大鼠,股动脉插管,腹腔注射安非他酮10 mg·kg^(-1),给药前和给药后5、10、30、60、120、180、240、360、480、600 min股动脉取血,测定安非他酮及其活性代谢产物浓度。结果血浆标准曲线在1～1000 ng·mL^(-1)(安非他酮、羟基安非他酮)、1～500 ng·mL^(-1)(赤式/苏式羟化安非他酮)范围内线性良好(r≥0.995),日内日间精密度均小于7%,绝对回收率为94.92%～117.57%,基质效应小于11.1%。安非他酮和羟基安非他酮药动学参数C_(max)分别为(1 543.33±241.94)、(22.40±7.66)ng·mL^(-1),T_(max)分别为(0.11±0.05)、(4.67±1.15)h,AUC_(0-10h)分别为(3 153.29±853.07)、(190.63±76.66)ng·h^(-1)·mL^(-1),半衰期分别为(2.63±0.79)、(29.76±17.95)h。苏式羟化安非他酮C_(max)、T_(max)、AUC_(0-10h)分别为(2.22±0.12)ng·mL^(-1)、(1.19±1.58)h、(9.95±6.56)ng·h^(-1)·mL^(-1)。结论建立的分析方法灵敏、准确,样品处理简便、快速,适合于血浆中安非他酮及其活性代谢产物的药动学研究。Objective To develop a method for simultaneous determination of bupropion(BUP) and its metabolites in rat plasma,and to explore its application in pharmacokinetic study. Methods The chromatographic separation was performed by using ultra performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS) on Agilent Zorbax SB-C8 column(4.6 mm× 100 mm ,3.5μm) in the mobile phase of water/methanol(5 mmol·L^-1 ammonium formate,pH 3.6).MS/MS detection was operated by multiple reaction monitoring in a positive mode with electrospray ion source.Ion-pairs for quantitative analysis were as follows: m/z240→m/z184(BUP), m/z256→ m/z238(hydroxybupropione,HB),m/z242→m/z168(erythrohydrobupropion/threohydrobupropion,EB/TB),m/z285→m/z154(diazepam,internal standard).Three female SD rats were selected and femoral artery was catheterized.Each rat received intraperitoneal injection of 10 mg· kg -1 BUP.Blood samples were collected before administrationand at 5,10,30,60,120,180,240,360,480 and 600 minutes after administration to measure the concentrations of BUP and its metabolites. Results The calibration curve obtained was linear( r ≥0.995) over the concentration ranges of 1-1000 ng·mL^-1 (BUP,HB) and 1-500 ng·mL^-1 (EB/TB).Inter-day and intra-day precisions were less than 7%.The absolute recoveries were 94.92%-117.57%and matrix effects were less than 11.1%.The C max ,T max ,AUC 0-10h and t1/2 were respectively(1 543.33± 241.94 )ng·mL^-1 ,(0.11±0.05)h,(3 153.29±853.07)ng·h^-1·mL^-1 and(2.63±0.79)h for BUP,and ( 22.40± 7.66)ng·mL^-1 ,(4.67±1.15)h,(190.63±76.66)ng·h^-1·mL^-1 and(29.76±17.95)h for HB. The C max ,T max and AUC 0-10h for TB were(2.22±0.12)ng·mL^-1 ,(1.19±1.58)h and (9.95±6.56)ng·h^-1 ·mL^-1 , respectively. Conclusion The analytical methodestablished in this study is sensitive,accurate,simple and fast for pharmacokinetic study of BUP and its metabolites in plasma.

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

分 类 号：R917[医药卫生—药物分析学]