A rapid and sensitive LC-MS/MS analysis of diapocynin in rat plasma to investigate in vitro and in vivo pharmacokinetics

Abstract

Natural products have been used traditionally for the treatment and prevention of diseases. Diapocynin is the analogue of the naturally occurring apocynin, a well-known constituent of Picrorhiza kurroa. It has antioxidant properties and also is effective in neuro-inflammatory diseases. To investigate in vitro and in vivo pharmacokinetics of diapocynin, a simple and sensitive method was developed and validated in rat plasma using high-performance liquid chromatography coupled with a mass spectrometer (LC-MS/MS). The developed and validated method requires low volume of the plasma (25 μL). The analytes were extracted by a simple protein precipitation method using acetonitrile. Chromatography resolution = n was achieved by using a Thermo Accucore C₁₈ (5 μ, 4.6 × 150 mm) column using a mobile phase of acetonitrile and ammonium acetate buffer (10 mM, pH 4.0). The present method was successfully applied for in vitro clearance (using rat liver microsomes) and in vivo oral pharmacokinetic studies in the Sprague Dawley rats.