高效液相色谱法测定血清中苯巴比妥、苯妥英、卡马西平、氯硝基安定的浓度

Determination of phenobarbital, phenytoin, carbamazepine and clonazepam in serum by HPLC

目的 :建立血清中苯巴比妥 (PB)、苯妥英 (PT)、卡马西平 (CBZ)、氯硝基安定 (CNZP)浓度测定的高效液相色谱法。方法 :以 PB、PT互为内标 ,ODS-Hypersil柱为分析柱 ,甲醇 -水 -β-环糊精 (4 5∶ 5 5∶ 0 .0 1)为流动相 ;流速为 0 .8ml/min,紫外检测波长 2 5 4nm。结果 :PB、PT、CBZ、CNZP分离良好 ,最低检测浓度分别为 0 .2 5、0 .5、0 .0 5、0 .0 15 μg/ml;线性范围分别为 2 .5～40、2 .5～ 40、1.2 5～ 2 0、0 .0 2～ 0 .3 2 μg/ml;相对回收率分别为 97.8%、10 2 .1%、10 2 .3 %、10 3 .3 % ;日内、日间 RSD均小于10 %。结论 :该法操作简便 ,结果可靠 。

AIM: to establish an assay method for the determination of phenobarbital(PB), phenytoin(PT), carbamazepine(CBZ) and clonazepam (CNZP) in serum by high performance liquid chromatography (HPLC). METHODS: Internal standards were phenobarbital and phenytoin. The analytical column was ODS Hypersil column. The mobile phase consisted of methanol water β cyclodextrin (45∶55∶0.01) and the flow rate was 0.8ml/min. The UV wavelength was 254nm. RESULTS: PB, PT, CBZ and CNZP were separated well. The lowest detection concentration of PB, PT, CBZ and CNZP were as following: 0.25, 0.5, 0.05 and 0.015 μg/ml; The linear relation was in the range of 2.5～40, 2.5～40, 1.25～20 and 0.02～0.32 μg/ml. Relative recovery rate were 97.8%, 102.1%, 102.3% and 103.3% respectively. The relative standard deviation (RSD) of within day and between day were all less than 10%. CONCLUSION: The method is found to be sensitive with constant result. The experiment will supply reference methods for the determination of PB, PT, CBZ and CNZP in serum.