RP-HPLC同时测定人血清中苯巴比妥、苯妥英、卡马西平的浓度

Determination of Phenobarbital,Phenytoin and Carbamazepine in Serum by RP-HPLC

目的 建立反相高效液相色谱法同时测定人血清中抗癫药苯巴比妥 (PB)、苯妥英 (PT)、卡马西平(CBZ)的浓度。方法 以巴比妥 (BB)为内标 ,样品用乙醚提取 ,45℃水浴吹干有机层 ,残留物用 15 0 μL甲醇复溶后进样。用反相柱YWGC18( 4.6mm× 3 0 0mm ,10 μm)分析 ,柱温 45℃ ,流动相甲醇 水 ( 5 5∶45 ) ,流速 0 .5mL/min ,紫外检测波长 2 3 0nm。结果 PB、PT、CBZ的保留时间分别为 11.77、17.5 3和 2 2 .2 9min ;检测限分别为 0 .2 5、0 .15、0 .0 5 μg/mL ;线性范围分别为 0 .5～ 80 (r =0 .9997)、0 .2 5～ 40 (r =0 .9996)、0 .2 5～ 2 0 (r =0 .9991) μg/mL ;相对回收率分别为 10 1.44 %、10 0 .0 7%、10 2 .0 9% ;日内RSD分别为 2 .77%、4.2 6%、1.5 4% ;日间RSD分别为 5 .95 %、3 .3 7%、5 .0 2 %。结论 此方法简便、结果准确 。

Purpose: To develop a method for simultaneously determining the concentrations of phenobarbital (PB), phenytoin (PT), carbamazepine (CBZ) in serum by RP-HPLC. Methods: The serum samples, added barbital as the internal standard, were extracted two times with aether. The organic phase was evaporated at 45°C. The residue was dissolved by methanol. YWG C 18 (4.6 mm x 300 mm, 10 μm) column was used; mobile phase: MeOH -H2O(55:45), flow rate 0.5 mL/min. The detective UV wavelength was 230 nm. Results: The parameters of PB, PT and CBZ were as follows respectively: tR 11.77, 17.53, 22.29 min; the lowest detection concentration 0.25, 0.15, 0.05 μg/mL; the linear relation was in the range of 0.5 - 80 (r = 0. 999 7), 0.25 - 40 (r = 0. 999 6), 0.25 - 20 (r = 0. 999 1) μg/mL. The average recoveries were 101.44%, 100.07%, 102.09%. The intra-day precision were 2.77%, 4.26%, 1.54%. The inter-day precision were 5.95%, 3.37%, 5.02%. Conclusions: The method was found to be accurate, convenient and effective. It is suitable for therapeutic drug monitoring of antiepileptic drugs and clinical pharmacokinetic study.