Rapid Analytical Method Development and Validation of RP-HPLC Method for Simultaneous Estimation of Diosmin and Hesperidin

Developing a single analytical method for estimation of individual drug from a multidrug composition is a very challenging task. A simple, rapid, precise, and reliable reverse phase HPLC method was developed for the separation and estimation of Diosmin and Hesperidin in bulk drug mix and pharmaceutical dosage forms. The estimation was carried out using Symmetry C18 (4.6×150mm, 5µ) column; mobile phase consisting of Methanol: Water (65:35v/v); the flow rate of 1mL/min and ultraviolet detection at 250 nm. All the drugs were properly resolved having run time of 8 minutes for Diosmin, and Hesperidin, respectively. The method was validated as a final verification of method development with respect to precision, linearity, accuracy, ruggedness, and robustness. The validated method was successfully applied to the commercially available pharmaceutical dosage form, yielding very good and reproducible result