Development of high-performance liquid chromatographic determination of salicylaldehyde isonicotinoyl hydrazone in rabbit plasma and application of this method to an in vivo study

• Published in: Journal of separation science Vol. 28; no. 12; pp. 1300 - 1306
• Main Authors: KOVARIKOVA, Petra, KLIMES, Jiri, STERBA, Martin, POPELOVA, Olga, MOKRY, Milan, GERSL, Vladimir, PONKA, Premysl
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.08.2005; WILEY‐VCH Verlag; Wiley
• Subjects: Aldehydes - blood; Analytical chemistry; Animals; Blood Chemical Analysis - methods; Blood Chemical Analysis - standards; Blood Chemical Analysis - statistics & numerical data; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography, High Pressure Liquid - methods; Chromatography, High Pressure Liquid - standards; Chromatography, High Pressure Liquid - statistics & numerical data; Drug Stability; Exact sciences and technology; HPLC; Hydrazones - blood; Iron Chelating Agents - analysis; Iron-chelator; Male; Other chromatographic methods; Pharmacokinetics; Quality Control; Rabbits; Salicylaldehyde isonicotinoyl hydrazone; SIH; Organic hydrazide; Solid phase extraction; Biological fluid; Chemical analysis; Intravenous administration; HPLC chromatography; Rabbit; Solvent extraction; Blood; Chemical enrichment; Blood plasma; Pyridine derivatives; Sample preparation; Siderophore; Hydrazone; Ionophore; HPLC; Quantitative analysis; Trace analysis; Validation; Chemical precipitation; Liquid liquid extraction; Benzene derivatives; Iron-chelator; Lagomorpha; In vivo; Vertebrata; Mammalia; SIH; Salicylaldehyde isonicotinoyl hydrazone; Animal; Chelating agent; Phenols; Iron Ions; Pharmacokinetics
• ISSN: 1615-9306; 1615-9314
• DOI: 10.1002/jssc.200500077

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 NaH2PO4·2 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 μg/mL), accuracy, precision, selectivity, recovery, and stability. A concentration of 0.26 μg/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.