Development and validation of reverse-phase high-performance liquid chromatographic method for determination of resveratrol in human and rat plasma for preclinical and clinical studies

Aim: The aim of the study was to develop and validate a simple and precise reverse-phase High Performance Liquid Chromatography (RP-HPLC) method for quantitative analysis of trans-resveratrol in human and rat plasma. Methods: HPLC method was developed by using Phenomenex Luna C₁₈ column (150 x 4.6 mm, 5 µm) and the optimized mobile phase comprised of acetonitrile/water in isocratic mode (30:70, v/v) with the flow rate of 1.0 mL/min. Trans-resveratrol was detected at a UV wavelength of 306 nm. Developed method was validated as per International Conference on Harmonization (ICH) M10 guidelines. Results: The proposed method was found simple, precise and linear with regression coefficient of 0.999 which could analyse the samples in nanograms levels with mean percent recovery in the acceptable range of 94.44 – 97.44%. The method was precise at the intra-day and inter-day levels as reflected by the relative standard deviation values (less than 3.36%). Trans resveratrol was found to be stable in plasma under different storage conditions. Conclusion: The present investigation demonstrated that the developed method was succesfully applied to accurately determine trans-resveratrol in human and rat plasma and therefore can be applicable for pre-clinical and clinical studies.