HPLC同时测定血清中苯妥英、苯巴比妥和卡马西平的浓度

DETERMINATION OF PHENYTION,PHENOBARBITONE AND CARBAMAZEPINE CONCENTRATION IN SERUM BY HPLC

目的 :建立同时测定微量血清中苯妥英、苯巴比妥和卡马西平浓度的方法。方法 :采用HPLC法 ,以巴比妥为内标同时测定。KromasilC18不锈钢色谱柱 ,流动相为甲醇 水 (5 7∶43) ,流速 0 .8ml·min 1,检测波长 2 5 4nm。结果 :检测浓度范围苯妥英为 4.0～ 5 0 .0 μg·ml 1(r=0 .9995 ) ,苯巴比妥 5 .0～ 6 0 .0 μg·ml 1(r =0 .9996 ) ,卡马西平为2 .5～ 2 0 .0 μg·ml 1(r =0 .9992 )。最低检测限分别为 1.0 μg·ml 1、2 .0 μg·ml 1和 0 .5 μg·ml 1;平均回收率分别为99.2 2 %、99.6 8%和 98.6 2 % ;日内和日间RSD均低于 5 % (n =5 )。结论 :该法准确、灵敏度高 ,重复性好 ,可作为

OBJECTIVE:To establish a method for rapid determination of phenytoin, phenobarbitone and carbamazepine in blood serum.METHODS:Using HPLC with barbitone as internal standard, the separation was performed on Kromasil C 18 column with methanol water (57∶43) as mobile phase. The flow rate was 0.8 ml·min 1 . The wavelength for detection was 254 nm.RESULTS:The linear range of phenytoin was 4.0 50 μg·ml 1 ( r =0.9995), phenobarbitone was 5.0 60 μg·ml 1 ( r =0.9996), carbamazepine was 2.5 20 μg·ml 1 ( r =0.9992), the lowest limit of detection was 1.0 μg·ml 1 , 2.0 μg·ml 1 and 0.5 μg·ml 1 . The average recoveries for phenytoin, phenobarbitone and carbamazepine were 99.22%,99.68%and 98.62%,respectively The within day and day to day RSD for 3 compounds were smaller than 5%( n =5).CONCLUSION:This method is accurate, with high sensitivity and good repeatability. It can be used as a routine method for TDM.