A percutaneous absorption study was conducted in rats in accordance with the OECD guidelines for an in vivo skin absorption test (OECD, 2004). Approximately 12 h prior to experimentation, rats were anaesthetized by isoflurane, and the dorsal skin covering an area of 6 × 6 cm2 was shaved with an electric clipper (Electric clipper Model 808, Daito Electric co., Osaka, Japan). The shaved skin surface was gently wiped with acetone to remove sebum. After overnight fasting, each formulation was applied to the shaved dorsal skin of rats covering an area of 4 × 4 cm2 (n = 5 per each formulation). The applied amount of each formulation was 1% of BCP, and the applied BCP dose was 0.262 mg/kg. At 12 h after topical application, the applied area was softly rinsed off with acetone to remove the remaining formulation on the skin. Blood samples (0.2 ml each) were collected from the jugular vein at 15, 30 min, and 1, 1.5, 2, 4, 6, 8, 10, 12, 24, 36, and 48 h after topical application. Plasma samples were harvested by centrifugation at 4,000 g for 10 min and stored at −70°C until analysis. These parameters included terminal half-life (t1/2), the area under the plasma concentration-time curve from time zero to the last observation time point (AUCall) and to infinity (AUCinf), apparent clearance (CL/F), apparent volume of distribution (Vz/F), and mean residence time (MRT). Maximum plasma concentration (Cmax) and time to reach Cmax (Tmax) were obtained directly from the observed data. The topical bioavailability (F) was calculated as F (%) = 100 (AUCtopical × Dose i.v.)/(AUCi.v.xDosetopical).

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