Determination of phenobarbital, phenytoin, carbamazepine and clonazepam in serum by HPLC
Wang Zengshou, Zhu Guanghui, Li Guangqian(The Affiliated second Hospital, Yuying Children's Hospital of Wenzhou Medical college, Wenzhou 325027)
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.
【CateGory Index】： R969