LC-MS法研究血浆中氧化苦参碱及其代谢物在犬体内的药代学

Pharmacokinetics of oxymatrine and its metabolite in beagle dogs by LC-MS

目的 :建立同时测定Beagle犬血浆中氧化苦参碱及其代谢物的LC MS分析法 ,研究氧化苦参碱及其代谢物药代学过程。方法 :采用LichrospherC18柱 (4.6mm× 2 5 0mm ,5 μm) ,柱温 2 5℃。流动相 10mmol·L-1醋酸铵水溶液 甲醇 (2 5∶75 ) ,流速 1mL·min-1;电喷雾离子化 (ESI)方式 ,采用选择性离子检测法 ,检测离子为正离子 ,氧化苦参碱SIM的离子是 [M +H]+ ,m/z 2 6 5 .1;苦参碱SIM的离子是 [M +H]+ ,m/z2 4 9 2。结果 :氧化苦参碱与苦参碱在 2～ 5 0 0 0ng·mL-1呈良好的线性关系 (r≥ 0. 9990 ) ,二者最小检出含量分别为 0. 6 ,0. 3ng·mL-1。日内和日间误差均小于 4.7% ,方法回收率大于 96. 5 % ;氧化苦参碱的T1/ 2 ,Tmax,Cmax,MRT ,AUC0→ 2 4h为 (5. 5± 1 5 8)h ,(1.0± 0 .30 )h ,(2 4 18 3± 970 78)ng·mL-1,(3.2± 0. 6 4 )h ,(5 797 4± 90 8 16 )ng·mL-1·h ,苦参碱的T1/ 2 ,Tmax,Cmax,MRT ,AUC0→ 2 4h为 (9.8± 2.77)h ,(1.9± 1.0 9)h ,(15 32. 4± 4 94 86 )ng·mL-1,(4 4± 1 97)h ,(5.5 30 5± 10 4 2.6 5 )ng·mL-1·h。结论 :氧化苦参碱和苦参碱LC MS定量法灵敏、快速、简单 ,专属性强 ;

Objective: To establish LC-MS method in the determination of oxymatrine and its metabolite in plasma and investigate their pharmacokinetics in beagle dogs. Method: Lichrospher C18 column (4.6 mm×250 mm, 5 μm )was used as the analytical columnm maintained at 25 ℃. The mobile phase was consisted of 10 mmol·L^(-1) CH_3COONH_4 and CH_3OH (25∶75). Flow rate was 1mL·min^(-1). Electrospray ionization(ESI)was carried out. The ESI ion source was set in positive ion polasity mode.The selective ion monitoring (SIM) was set at m/z 265.1 and 249.2. Result: The linearity ranged from 2 to 5 000 ng·mL^(-1)(r=0.999 1). The detectction of oxymatrine and its metabolite were 0.6 and 0.3 ng·mL^(-1).The RSD(%) within day and between day was less than 4.7 %. The recovery of this method was more than 96.5%. The disposition was conformed to a two-compartment model. The T_(1/2), T_(max), C_(max), MRT, AUC_(0→24 h) of oxymatrine were ((5.5±)1.58) h,(1.0±0.30) h,(2 418.3±970.78) ng·mL^(-1),(3.2±0.64) h,(5 797.4±908.16) ng·mL^(-1)·h accordingly. The corresponding T_(1/2), T_(max), C_(max), MRT, AUC_(0→24h) of matrine were(9.8±2.77) h,(1.9±1.09) h,(1 532.4±494.86) ng·mL^(-1),(4.4±1.97) h,((5 530.5±)1 042.65) ng·mL^(-1)·h. Conclusion: This assay was highly senstive, rapid, simple and specific enough for determining concentrations of oxymatrine and its metabolite matrine in plasma of beagle dog.