title: Analysis of new potential anticonvulsant compounds in mice brain tissue by SPE/HPLC/DAD.
abstract: This paper describes a novel reversed-phase high performance liquid chromatography (RP-HPLC) with photo-diode array detection (DAD) method for the determination of three new derivatives of 4-alkyl-5-(3-chlorophenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione with different antiepileptic activity in the brains of mice treated with the doses of 300mgkg(-1) of body weight. Samples were prepared by solid-phase extraction (SPE) method using BAKERBOND™ spe Octadecyl (C(18)) and analyzed by the use of an isocratic elution mode over an Zorbax Extend-C18 column (150mm×4.6mm I.D., 5-μm, Agilent Technologies). The mobile phase consisted of 80% methanol (for compound TP-315) and 85% acetonitrile (for compound TP-321) for 80% 2-propanol (for TP-323) at a flow rate of 1.0mLmin(-1) and 0.5mLmin(-1) in the last case. Gradient elution mode was also proposed for all examined analytes in mixture with common antiepileptic drugs: carbamazepine, phenobarbital and phenytoin in view of possible synergistic activity. Photodiode-array investigations of the peaks after degradation studies indicate the stability of the compounds under conditions proposed for sample preparation procedure. Linear coefficients of correlation (r(2)) were >0.995 for all analytes. The proposed strategy gives extraction yields higher than 95% with the intra- and inter-day relative standard deviation lower than 3% and 5%, respectively. This method was applied to the analysis of brain tissue of mice treated with investigated compounds. Obtained results enable to explain the differences in their pharmacological activity.
authors.affiliation: Department of Analytical Chemistry, Medical University of Lublin, Chodźki 4A, 20-093 Lublin, Poland. j.flieger@umlub.pl
authors.forename: Jolanta
authors.initials: J
authors.lastname: Flieger
authors.forename: Magdalena
authors.initials: M
authors.lastname: Pizoń
authors.forename: Tomasz
authors.initials: T
authors.lastname: Plech
authors.forename: Jarogniew J
authors.initials: JJ
authors.lastname: Luszczki
mesh_terms: D000818:Animals; D000927:Anticonvulsants / Q000032:analysis* / Q000737:chemistry; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D051379:Mice; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000379:methods*; D014230:Triazoles / Q000032:analysis* / Q000737:chemistry
chemical_list: D000927:Anticonvulsants; D014230:Triazoles; C045575:1,2,4-triazole