Dissertation committee:
Σ. Αθανασέλης, Καθηγητής, Χ. Σπηλιοπούλου, Καθηγήτρια, Ι. Παπουτσής, Επικ. Καθηγητής
Summary:
The aim of this thesis was the development, optimization and validation of gas
chromatography-mass spectrometric methods for the simultaneous determination of
antiepileptic drugs, in plasma and whole blood. During the method development,
the necessity appeared for separation of the antiepileptics according to their
chemical behavior. So, two methods were actually developed and validated, one
for the determination of the nine alkaline antiepileptics (ethosuximide,
carbamazepine, lacosamide, lamotrigine, levetiracetam, oxcarbazepine,
phenobarbital, phenytoin and topiramate) and two metabolites
(10,11-epoxide-carbamazepine and 10-hydroxy-carbazepine), and one for the
antiepileptics with acidic (valproic acid and tiagabine) and zwitterionic
(gabapentin, pregabalin and vigabatrin) behavior. The sample pretreatment
included protein precipitation with acetonitrile, solid phase extraction and
derivatization using MTBSTFA with 1% TBDMSCl at 70oC for 30min.
Levetiracetam-d6 was used as internal standard for the determination of
alkaline antiepileptics, whereas gabapentin-d10 was used for the acidic and
zwitterionic antiepileptics. The validation of the methods included the
evaluation of: selectivity, specificity and linearity, limits of detection and
quantification, intra- and inter-day accuracy and precision, absolute recovery
of the methods and the stability of analytes in spiked blood samples. The
developed methods were applied for therapeutic antiepileptic drug monitoring
and for the toxicological investigation of clinical and forensic cases.
Keywords:
epilepsy, antiepileptic drugs, plasma, blood, forensic and clinical toxicology