| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1212171 | Journal of Chromatography B | 2015 | 4 Pages |
•The validated HPLC method was suitable for SK5474 quantification in plasma.•SK5474 was stable under various conditions, whether plasma or stock solutions.•The F value of SK5474 showed good bioavailability for entering preclinical studies.•The major elimination is thought to be hepatic and gastrointestinal first-pass.
In this study, a method for the quantitation of SK5474 in rat plasma was developed and validated using high performance liquid chromatography (HPLC). The plasma samples were prepared by deproteinization, and sildenafil was used as an internal standard. Chromatographic separation was achieved using a reversed-phase (C18) column. The mobile phase, 0.02 M ammonium acetate buffer:acetonitrile (48:52, v/v), was run at a flow rate of 1.0 mL/min, and the column eluent was monitored using an ultraviolet detector at 254 nm at room temperature. The retention times of sildenafil and SK5474 were approximately 5.8 and 7.2 min, respectively. The quantitation limit of SK5474 in rat plasma was 0.03 μg/mL. Pharmacokinetic parameters of SK5474 was evaluated after intravenous (i.v.; at doses of 15 mg/kg) and oral (p.o.; at doses of 30 mg/kg) administration of SK5474 in rats. After p.o. administration (30 mg/kg) of SK5474, F (fraction absorbed) value was approximately 46.0%.
