title: Development of high-performance liquid chromatographic determination of salicylaldehyde isonicotinoyl hydrazone in rabbit plasma and application of this method to an in vivo study.
abstract: An analytical methodology appropriate for the determination of the novel drug candidate salicylaldehyde isonicotinoyl hydrazone (SIH) in rabbit plasma has been developed and validated. Desirable chromatographic separation was achieved on a C18 column employing a mixture of phosphate buffer (0.01 M NaH2PO4 x 2 H2O with 2 mM EDTA, pH 6.0) and methanol (53:47; v/v) as the mobile phase. In order to develop a suitable sample preparation procedure, different methods have been tested (solid-phase extraction, liquid-liquid extraction, and protein precipitation). Protein precipitation using 0.1 M HClO4 and acetonitrile allowed the highest recoveries of the analyte to be reproducibly attained. The analytical methodology developed in this study was validated with respect to linearity (0.26-30.0 microg/mL), accuracy, precision, selectivity, recovery, and stability. A concentration of 0.26 microg/mL was determined as the LLOQ. The chromatographic method was applied to a preliminary plasma pharmacokinetic study. This study has provided the first information about the concentrations of SIH in plasma of a living subject. These results could have a significant impact on further progress in the development of this promising compound.
authors.affiliation: Department of Pharmaceutical Chemistry and Drug Control, Faculty of Pharmacy in Hradec Králové, Charles University in Prague; Hradec Králové, Czech Republic. kovarikova@faf.cuni.cz
authors.forename: Petra
authors.initials: P
authors.lastname: Kovaríková
authors.forename: Jirí
authors.initials: J
authors.lastname: Klimes
authors.forename: Martin
authors.initials: M
authors.lastname: Stĕrba
authors.forename: Olga
authors.initials: O
authors.lastname: Popelová
authors.forename: Milan
authors.initials: M
authors.lastname: Mokrý
authors.forename: Vladimír
authors.initials: V
authors.lastname: Gersl
authors.forename: Premysl
authors.initials: P
authors.lastname: Ponka
mesh_terms: D000447:Aldehydes / Q000097:blood*; D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods / Q000592:standards / Q000706:statistics & numerical data; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D004355:Drug Stability; D006835:Hydrazones / Q000097:blood*; D007502:Iron Chelating Agents / Q000032:analysis; D008297:Male; D011786:Quality Control; D011817:Rabbits
chemical_list: D000447:Aldehydes; D006835:Hydrazones; D007502:Iron Chelating Agents; C413684:salicylaldehyde isonicotinoyl hydrazone