Determination of donepezil in spiked rabbit plasma by high-performance liquid chromatography with fluorescence detection

The aim of this paper is to develop sensitive, accurate, reproducible and robust RP-HPLC with fluorescence detection for estimation of donepezil (DZ) in rabbit plasma using silodosin as the internal standard (IS). The prepared samples were quantified on reversed phase column Luna C(18(2)) (150 × 4.6 mm i.d., 5 µm particle size) operated at room temperature using the mobile phase consisting of methanol: 0.1% acetic acid (50 : 50, v/v) at a flow rate of 1 ml min(−1). The method was fully validated according to bioanalytical validation guidelines of FDA in terms of system suitability, selectivity, sensitivity, precision and stability. It was found that the increase in peak areas followed the increase of DZ concentration in the range of 2.56–200.00 ng ml(−1) with LOD of 0.85 ng ml(−1). The method was successfully applied for the determination of DZ in rabbit plasma using manual shaking dispersive liquid–liquid microextraction.