DETERMINATION AND VALIDATION OF NARINGIN IN RAT PLASMA BY RP-HPLC

A simple, accurate isocratic reverse phase high performance liquid chromatography (RP-HPLC) method was developed for the determination of the plasma concentration of naringin, a flavanone glycoside in rat plasma. An assay procedure involved simple liquid–liquid extraction of naringin from plasma directly into methanol. The organic layer was separated, filtered and injected onto a Phenomenex-C18 column (4.6 × 250 mm, 5 μm). The mobile phase acetonitrile and water (30:70, V/V) was used at a flow rate of 1.0 mL/min for the effective separation of naringin. The detection of the analyte peak was achieved by monitoring the eluate using a UV detector at 283 nm. The peak area of analyte was used for quantification of plasma samples. The retention time of naringin was 4.7 min. The standard curve for naringin was linear (r2 = 0.9987) in the concentration range 100-2000 ng/mL. The absolute recoveries of naringin were in the range 85–115% from rat plasma. The lower limit of quantification (LLOQ) of naringin was 30 ng/mL. The intra and inter-assay precisions in the measurement of quality control samples of 100, 400, 900 and 1600 ng/mL, were in the range of 0.19-5.44% and 0.27-2.35% relative standard deviation (RSD), respectively. Accuracy in the measurement of quality control samples was in the range 98.40-107.9% of the spiked nominal values. The plasma concentration of naringin was increased with increase in test dose levels. From the results, maximum plasma concentration of naringin was found to be 289.03 and 757.3 ng/mL with 15 and 30 mg/kg, oral doses, respectively at 2 h. This method is simple and helps in determination of naringin in rat plasma.