Analytical method development and validation for assay of Diosmin and Hesperidin in combined tablet dosage form by RP- HPLC

Abstract

The combination of Diosmin and Hesperidin can be used as capillary stabilizing and antihemorrhoidal agent. A rapid, simple, precise and cost effective and reversed-phase high performance liquid chromatography (RP-HPLC) method has been developed and validated for the simultaneous determination of Diosmin and Hesperidin in pharmaceutical formulations. Separation of both Diosmin and Hesperidin was achieved within 8 min with required resolution. Chromatographic separation was achieved on a waters symmetry C18 (100mm x 4.6mm) 2.6µm using a mobile phase consisting of 0.1% ortho phosphoric acid and methanol in the ratio of 60:40 at a flow rate of 1.0 ml/min. The detection was made 280 nm at nm and the retention time of Diosmin and Hesperidin were 6.049 and 8.560 minutes respectively.