Extraction and Characterization of Hesperidine Present in Natural and Polyherbal formulation

Hesperidine, a bioflavonoid, is an abundant and inexpensive by-product of Citrus cultivation. A deficiency of this substance in the diet has been linked with abnormal capillary leakiness as well as pain in the extremities causing aches, weakness and night leg cramps. No signs of toxicity have been observed with the normal intake of hesperidine or related compounds. Hesperidine was extracted from the dried peel of Citrus limetta which belongs to the family Rutaceae. An amount of 1kg air-dried and powdered peel was extracted to get 10gms of hesperidine which was a white brownish solid. Analysis of isolated hesperidine along with standard Hesperidine and marketed ayurvedic polyherbal formulation were identified by using IR spectroscopy, UV Spectroscopy, Thin Layer Chromatography (TLC) and UFLC. IR characterization showed that the standard, isolated and ayurvedic formulation were same. In UV spectroscopy all the components showed specific absorbance at 283.5nm. The Rf values for isolated, standard and marketed ayurvedic formulation were found to be 0.710, 0.708 and 0.702 respectively. A reversed phase UFLC method was also developed for determination and quantification of hesperidine. The method involved the use of a Phenomenex Luna 5u C18 column, a mixture of water-Acetonitrile-methanol (50:25:25 v/v/v) as a mobile phase in an isocratic mode at a flow rate of 1 ml/min and UV detection at 283.5 nm.