Hplc method for the determination of hesperidin and diosmine in flavonoid extract and capsules

ABSTRACT

A high performance liquid chromatographic procedure was developed for the determination of hesperidin and diosmine in the flavonoid extract of rutaceae and in dosage forms. The method utilized a reversed phase chromatography with UV detection. The mobile phase comprised of 38% methanol and 62% 0.005 M heptane sulfonic acid sodium salt pumped at a flow rate of 1.5 ml/min and the column was at ambient temperature. The detection wavelength was 278 nm and the quantitation was performed by measuring the peak area. It was also determined that the excipients in the commercial soft gelatin capsule preparations containing these flavonoids did not interfere with the assay. The method was linear in the range 5-40 mug/ml. In all cases, the coefficient of variation did not exceed 2%