An HPLC–ultraviolet detection method for the determination of Z24 in mouse whole blood and its application to pharmacokinetic studies

A sensitive and reproducible high-performance liquid chromatography (HPLC)–UV method for the determination of Z24, a tumorigenesis and angiogenesis inhibitor, has been developed and validated in mouse whole blood. Blood samples were extracted with ether, evaporated, and the residue was reconstituted in mobile phase. An aliquot was separated by isocratic reversed-phase HPLC on a Hypersil ODS-2 column and quantified using UV detection at 390 nm. The mobile phase was 50% (v/v) acetonitrile/water with a flow rate of 0.8 ml/min. A linear curve over the concentration range of 0.05–6 μg/ml (r2 = 0.9976) was obtained. The coefficient of the variation for the intra- and inter-day precision ranged from 3.0 to 10.9% and 5.7 to 10.3%, respectively. The absolute recovery of Z24 was 89.2–108.5%. The method is simple, economical and sufficient for in vivo pharmacokinetic studies on Z24. Nonlinear pharmacokinetics was found in mice at doses from 20 to 80 mg/kg.