兔血浆中霉酚酸酯的活性代谢物霉酚酸的HPLC测定及其药代动力学研究

目的建立霉酚酸兔血药浓度测定方法 ,并进行了灌胃后药代动力学研究。方法采用HPLC 二极管阵列检测器测定血药浓度。血样采用乙醚萃取 ;色谱柱 :HypersilODS2 (2 0 0mm×4 6mm ,5 μm) ;流动相 :0 0 5mol·L-1KH2 PO4 CH3 CN(5 5∶45 ,pH 2 5 ) ;检测波长 :2 1 5nm ;流速 :1 0mL·min-1。结果霉酚酸血药浓度线性范围为 0 1 6～ 1 9 98μmol·L-1(r =0 9996,n =8) ,血浆最低检测浓度为 0 0 3 μmol·L-1,高、中、低 3种浓度平均回收率分别为 :(1 0 3 2 5± 6 3 2 ) %、(99 63± 2 75 ) %、(1 0 2 47± 3 1 8) % ,兔灌胃后主要的药代动力学参数分别为 :达峰浓度cmax为(8 2 7± 6 1 2 ) μmol·L-1,达峰时间tmax为 (0 49± 0 1 7)h ,消除半衰期t1/ 2 为 (3 1 2± 0 94)h ,药时曲线下面积AUC0 -t为 (1 3 0 8± 9 0 8) μmol·h·L-1,AUC0 -∞ 为 (1 3 80± 8 93 )mg·h·L-1。结论为霉酚酸的血药浓度监测及药动学研究提供了实验方法。

Determination of mycophenolic acid,the active metabolite of mycophenolate mofetil in rabbit plasma by HPLC and pharmacokinetic study

Objective To establish a method for determining mycophenolic acid(MPA) in rabbit plasma.Method A high performance liquid chromatographic assay with diode array detection was developed.The detection wavelength was set at 215 nm .The plasma sample was extracted with diethyl ether.The sample was separated on a Hypersil ODS2 column ( 200 mm × 4.6 mm , 5 μm ),with 0.05 mol·L -1 KH_2PO_4-CH_3CN(55∶45,pH 2.5) as the mobile phase.The flow rate was 1.0 mL·min -1 .Results The calibration curve of MPA was linear between 0.16～ 19.98 μmol·L -1 ( r = 0.999 6 , n =8).The detection limit was 0.03 μmol·L -1 .The mean recoveries of high,medium and low concentrations were (103.25±6.32)%,(99.63±2.75)% and (102.47±3.18)%,respectively.The pharmacokinetic parameters after ig were as follows: c _ max (8.27±6.12)μmol·L -1 , t _ max (0.49±0.17)h, t _ 1/2 (3.12±0.94)h,AUC_ 0- t (13.08±9.08)μmol·h·L -1 ,AUC_ 0→∞ (13.80±8.93)μmol·h·L -1 .Conclusion The present study provides a reliable quantitative method for pharmacokinetic study and monitor of MPA.