Tissue Distribution Study of Apocynin By RP-HPLC With UV Detector

Abstract

Background Apocynin (4-hydroxy-3-methoxyacetophenone), an important active ingredient contained in root of Picrorhiza kurroa Royle, has been widely investigated as an antioxidative and anti-inflammatory agent in kinds of disease models, and exerted certain efficacy deserving further research. However, little was known about the disposal process of apocynin in vivo, which is important information required in drug research and development.

Aim In this work, a tissue distribution study of apocynin was performed in Sprague-Dawley (SD) male rats to help further understanding well the disposition of apocynin in vivo.

Methods A simple HPLC-UV method was developed for measurement of apocynin concentration in rat tissues. A mixture of water-methanol (47:53, v/v) was used as solvent system, the flow rate was 1 mL/min, and the detected wavelength was 279 nm. The method was validated and applied to the tissue distribution study of a single bolus intravenous administration of apocynin in SD male rats.

Results The developed HPLC-UV method showed good specificity, precision, accuracy and extraction recovery. The good linearity was achieved within 0.8-32 ?g/mL in tissues including heart, liver, spleen, lung, kidney and brain. The lower limit of quantification (LLOQ) was 0.8 ?g/mL. The method was well used in the study of tissue distribution of apocynin. The results demonstrated that apocynin was distributed fast into the tested tissues and reached peak concentration at 5 min after injection. Apocynin was mainly distributed in liver, kidney and lung within 15 minutes after administration, and eliminated from the tissues with no sample be detected more than 2 h after dose.

Conclusion The tissue distribution behavior of apocynin is similar as its pharmacokinetic behavior in plasma, which showed a short half-life and fast elimination in SD rats. This work provided scientific reference for further research of apocynin.