title: First HPLC-UV method for rapid and simultaneous quantification of phenobarbital, primidone, phenytoin, carbamazepine, carbamazepine-10,11-epoxide, 10,11-trans-dihydroxy-10,11-dihydrocarbamazepine, lamotrigine, oxcarbazepine and licarbazepine in human plasma.
abstract: A sensitive and fast high-performance liquid chromatographic method coupled with ultraviolet detection is herein reported for the simultaneous determination of human plasma concentration of six antiepileptic drugs frequently used in clinical practice [phenobarbital (PB), primidone (PRM), phenytoin (PHT), carbamazepine (CBZ), lamotrigine (LTG), oxcarbazepine (OXC)] and some of their main metabolites, carbamazepine-10,11-epoxide (CBZ-E), 10,11-trans-dihydroxy-10,11-dihydrocarbamazepine (trans-diol) and licarbazepine (Lic). Sample preparation consisted of a deproteinization step with methanol followed by a solid-phase extraction procedure. Chromatographic separation was achieved in approximately 15 min on a reversed-phase C18 column using a mobile phase composed by water-methanol-acetonitrile-triethylamine (68.7:25:6:0.3, v/v/v/v; pH 6.5) pumped isocratically at 1.0 mL/min. The detector was set at 237 nm. Calibration curves were linear with regression coefficients greater than 0.992 over the concentration ranges 0.25-100 μg/mL for PB, 0.4-50 μg/mL for PRM, 0.5-50 μg/mL for PHT, 0.1-50 μg/mL for CBZ, LTG and CBZ-E, 0.1-25 μg/mL for OXC, 0.25-10 μg/mL for trans-diol and 0.15-80 μg/mL for Lic. Inter- and intra-day imprecision did not exceed 12.15% and inaccuracy was within ±14.91%. Absolute mean recoveries ranged from 78.49 to 101.04% and no interferences were observed at the retention times of the analytes and internal standard (ketoprofen). This bioanalytical method was successfully applied to real plasma samples from epileptic patients and it seems to be a suitable tool for routine therapeutic drug monitoring and also to support other clinical pharmacokinetic-based studies.
authors.affiliation: Laboratory of Pharmacology, Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal.
authors.forename: Ana
authors.initials: A
authors.lastname: Serralheiro
authors.forename: Gilberto
authors.initials: G
authors.lastname: Alves
authors.forename: Ana
authors.initials: A
authors.lastname: Fortuna
authors.forename: Marília
authors.initials: M
authors.lastname: Rocha
authors.forename: Amílcar
authors.initials: A
authors.lastname: Falcão
mesh_terms: D000927:Anticonvulsants / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D002220:Carbamazepine / Q000031:analogs & derivatives / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016903:Drug Monitoring; D004355:Drug Stability; D006801:Humans; D000077213:Lamotrigine; D057230:Limit of Detection; D016014:Linear Models; D010634:Phenobarbital / Q000031:analogs & derivatives / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D010672:Phenytoin / Q000031:analogs & derivatives / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D015203:Reproducibility of Results; D014227:Triazines / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D000927:Anticonvulsants; D014227:Triazines; D002220:Carbamazepine; D010672:Phenytoin; D000077213:Lamotrigine; D010634:Phenobarbital