高效液相色谱法同时测定血清中苯妥英、卡马西平的浓度

Determination of phenytoin and carbamazepine in blood serum by RP-HPLC with UV detection

目的 :建立HPLC法同时测定血清苯妥英 (PHT)与卡马西平 (CBZ)浓度。方法 :以C18反相柱为色谱柱 ,甲醇 水 (5 5∶45 )为流动相 ,检测波长 2 40nm ;用乙酸乙酯作为提取剂。结果 :PHT、CBZ平均回收率各为 99.5 %和 10 0 .5 % ,日内和日间RSD均低于 6 .6 % (n =5 ) ;分析方法的定量测定下限 :PHT为 1.2 μ犂·ml-1,CBZ为 0 .2 μ犂·ml-1。分析方法的检测下限 :PHT为 0 .4μ犂·ml-1;CBZ为 0 .1μ犂·ml-1。PHT在 5～ 40 μ犂·ml-1浓度范围线性关系良好 ,r=0 .998;CBZ在 2 .5～ 2 5 μ犂·ml-1浓度范围线性关系良好 ,r=0 .998。结论 :方法灵敏、准确 ,可用于PHT和CBZ临床血药浓度监测。

OBJECTIVE:To develop an HPLC method for quantitative determination of phenytoin and carbamazepine in blood serum. METHODS:Blood serum samples were extracted with hexane to remove lipid and interfering substances , phenytoin (PHT) and carbamazepine (CBZ) were then extracted with ethyl acetate. The residues dissolved were analyzed with a reverse phase HPLC system (C 18 column, 4.6 mm×250 mm; Mobile phase, MeOH H 2O( 55∶45); UV detection, 240 nm).RESULTS:The average recoveries for PHT and CBZ were 99.5% and 100.5% , respectively. The within day and day to day relative standard deviations for both compounds were all smaller than 6.6% (n = 5) . The calibration curves for both compounds had good linearity, r = 0.998 for PHT within a concentration range of 5～40 μg·ml -1 , and r = 0.998 for CBZ within a concentration range of 2.5 ～25 μg·ml -1 . The limits of detection (LODs) for PHT and CBZ were 0.4 μg·ml -1 and 0.1 μg·ml -1 , and the limits of quantitation (LOQs) for PHT and CBZ were 1.2 μg·ml -1 and 0.2 μg·ml -1 , respectively.CONCLUSIONS:The method provides a sensitive, accurate, precise and reliable analytical procedure for clinical monitoring of blood serum PHT and CBZ.[