A chiral HPLC-UV method for the quantification of dibenz[b,f]azepine-5-carboxamide derivatives in mouse plasma and brain tissue: Eslicarbazepine acetate, carbamazepine and main metabolites
✍ Scribed by Ana Fortuna; Joana Bicker; Gilberto Alves; Amílcar Falcão; Patrício Soares-da-Silva
- Publisher
- John Wiley and Sons
- Year
- 2011
- Tongue
- English
- Weight
- 331 KB
- Volume
- 34
- Category
- Article
- ISSN
- 1615-9306
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✦ Synopsis
Abstract
For the first time, a selective and sensitive chiral HPLC‐UV method was developed and fully validated for the simultaneous quantification of eslicarbazepine acetate (ESL), carbamazepine (CBZ), S‐licarbazepine (S‐Lic), R‐licarbazepine (R‐Lic), oxcarbazepine (OXC) and carbamazepine‐10,11‐epoxide (CBZ‐E), in mouse plasma and brain homogenate supernatant. After the addition of chloramphenicol as the internal standard, samples were processed using an SPE procedure. The chiral chromatographic analysis was carried out on a LiChroCART 250‐4 ChiraDex column, employing a mobile phase of water and methanol (88:12, v/v) pumped at 0.9 mL/min and the UV detector set at 235 nm. The assay was linear (r^2^≥0.995) for ESL, CBZ, OXC, S‐Lic, R‐Lic and CBZ‐E in the range of, respectively, 0.2–4, 0.4–30, 0.1–60, 0.2–60, 0.2–60 and 0.2–30 μg/mL, in plasma, and of 0.06–1.5 μg/mL for ESL, 0.12–15 μg/mL for CBZ and CBZ‐E and 0.06–15 μg/mL for OXC and both licarbazepine (Lic) enantiomers in brain homogenate supernatant. The overall precision was within 8.71% and accuracy ranged from −7.55 to 8.97%. The recoveries of all the compounds were over 92.1%. Afterwards, the application of the method was demonstrated using real plasma and brain samples obtained from mice administered simultaneously with ESL and CBZ.