title: Liquid chromatography separation of the chiral prodrug eslicarbazepine acetate and its main metabolites in polar organic mode. Application to their analysis after in vitro metabolism.
abstract: A LC method using a chiral stationary phase (CSP) with cellulose tris(3-chloro-4-methylphenylcarbamate) as chiral selector in polar organic mode (POM) was developed for the separation of the biopharmaceutic classification system (BCS) class II chiral prodrug eslicarbazepine acetate (ESL) and its main metabolites, namely eslicarbazepine, its optical antipode, (R)-licarbazepine, and the achiral oxcarbazepine (OXC). The percentage of methanol (MeOH) in the mobile phase containing acetonitrile (ACN) as the main solvent was found to significantly influence analyte retention and resolution. A reversal of elution order of OXC and (R)-licarbazepine was observed, depending on the MeOH percentage in the mobile phase. The optimized mobile phase consisted of ACN/MeOH/acetic acid/diethylamine (95/5/0.2/0.07; v/v/v/v). The potential of this chemo- and enantioselective LC method combined with solid-phase extraction (SPE) was then evaluated for in vitro metabolism studies using ESL as a model case. Only eslicarbazepine could be detected after incubation of ESL in human liver microsome systems.
authors.affiliation: Laboratory for the Analysis of Medicines, Dept. of Pharmacy, CIRM, University of Liège, CHU, B36, B-4000 Liege, Belgium. Electronic address: acservais@ulg.ac.be.
authors.forename: Anne-Catherine
authors.initials: AC
authors.lastname: Servais
authors.affiliation: Laboratory for the Analysis of Medicines, Dept. of Pharmacy, CIRM, University of Liège, CHU, B36, B-4000 Liege, Belgium.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Janicot
authors.affiliation: Laboratory for the Analysis of Medicines, Dept. of Pharmacy, CIRM, University of Liège, CHU, B36, B-4000 Liege, Belgium.
authors.forename: Arnold
authors.initials: A
authors.lastname: Takam
authors.affiliation: Laboratory for the Analysis of Medicines, Dept. of Pharmacy, CIRM, University of Liège, CHU, B36, B-4000 Liege, Belgium.
authors.forename: Jacques
authors.initials: J
authors.lastname: Crommen
authors.affiliation: Laboratory for the Analysis of Medicines, Dept. of Pharmacy, CIRM, University of Liège, CHU, B36, B-4000 Liege, Belgium.
authors.forename: Marianne
authors.initials: M
authors.lastname: Fillet
mesh_terms: D019342:Acetic Acid; D000097:Acetonitriles; D002220:Carbamazepine / Q000031:analogs & derivatives / Q000737:chemistry / Q000302:isolation & purification; D002482:Cellulose / Q000031:analogs & derivatives / Q000737:chemistry / Q000302:isolation & purification; D002853:Chromatography, Liquid / Q000379:methods; D003984:Dibenzazepines / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D004048:Diethylamines; D006801:Humans; D008862:Microsomes, Liver / Q000737:chemistry; D000078330:Oxcarbazepine; D048448:Phenylcarbamates / Q000737:chemistry / Q000302:isolation & purification; D011355:Prodrugs / Q000302:isolation & purification* / Q000378:metabolism; D052616:Solid Phase Extraction; D012997:Solvents; D013237:Stereoisomerism
chemical_list: D000097:Acetonitriles; D003984:Dibenzazepines; D004048:Diethylamines; D048448:Phenylcarbamates; D011355:Prodrugs; D012997:Solvents; C538979:cellulose tris(3-chloro-4-methylphenylcarbamate); D002220:Carbamazepine; D002482:Cellulose; C034281:diethylamine; C416835:eslicarbazepine acetate; D019342:Acetic Acid; C571001:eslicarbazepine; D000078330:Oxcarbazepine; C428744:10,11-dihydro-10-hydroxy-5H-dibenz(b,f)azepine-5-carboxamide; C032159:acetonitrile
keywords: Chiral stationary phase; Eslicarbazepine acetate; In vitro metabolites; Liquid chromatography