Development and validation of an HPLC-UV method for the simultaneous quantification of carbamazepine, oxcarbazepine, eslicarbazepine acetate and their main metabolites in human plasma
2010
Fortuna, Ana | Sousa, Joana | Alves, Gilberto | Falcão, Amílcar | Soares-da-Silva, Patrício
For the first time, a simple, selective and accurate high-performance liquid chromatography method with ultraviolet detection was developed and validated to quantify simultaneously three structurally related antiepileptic drugs; carbamazepine, oxcarbazepine, and the recently launched eslicarbazepine acetate and their main metabolites, carbamazepine-10,11-epoxide, 10,11-trans-dihydroxy-10,11-dihydro-carbamazepine, and licarbazepine. The method involves a solid-phase extraction and a reverse-phase C18 column with 5 cm length. The mobile phase consisting of water, methanol, and acetonitrile in the ratio 64:30:6 was selected as the best one and pumped at 1 mL/min at 40 °C. The use of this recent column and an aqueous mobile phase instead of buffers gives several advantages over the method herein developed; namely the fact that the chromatographic analysis takes only 9 min. The method was validated according to the guidelines of the Food and Drug Administration, showing to be accurate (bias within ±12%), precise (coefficient variation <9%), selective and linear (r ² > 0.997) over the concentration range of 0.05-30 μg/mL for carbamazepine; 0.05-20 μg/mL for oxcarbazepine; 0.15-4 μg/mL for eslicarbazepine acetate; 0.1-30 μg/mL for carbamazepine-10,11-epoxide; 0.1-10 μg/mL for 10,11-trans-dihydroxy-10,11-dihydro-carbamazepine, and 0.1-60 μg/mL for licarbazepine. It was also shown that this method can adequately be used for the therapeutic drug monitoring of the considered antiepileptic drugs, carbamazepine, oxcarbazepine, eslicarazepine acetate, and their metabolites.
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