Development and Validation of UV Spectrophotometric Method for Simultaneous Estimation of Hesperidin and Diosmin in the Pharmaceutical Dosage Form

A simple, rapid, precise and highly selective spectrophotometric method was developed for simultaneous estimation of Hesperidin and Diosmin in tablet dosage form. This method, involves the measurement of absorbances of Hesperidin and Diosmin at the wavelengths of 285？nm ( of Hesperidin) and 268？nm ( , of Diosmin). The UV spectra’s of Hesperidin and Diosmin prepared in different solvents water, methanol, and acetonitrile and 0.2？N sodium hydroxide were recorded. These two drugs showed good absorbances when dissolved in 0.2？N NaOH. Hence 0.2？N NaOH was selected as the solvent for the method. Two wavelengths 285 and 268？nm were selected which are of two drugs Hesperidin and Diosmin, respectively. Different concentrations of Hesperidin (5–50？μg/mL) and Diosmin (2–24？μg/mL) and a mixture of Hesperidin and Diosmin were prepared, scanned and absorbances were noted at the two wavelengths were fixed for the study. The method showed good reproducibility and recovery with % RSD less than 2. The LOD of Hesperidin and Diosmin was found to be 0.139？μg/mL and 0.048？μg/ml and LOQ of Hesperidin and Diosmin was found to be 0.42？μg/mL and 0.147？μg/mL, respectively. Thus the proposed method was found to be rapid, specific, precise, accurate and cost effective quality control tool for the routine analysis of Hesperidin and Diosmin in bulk and combined dosage form. 1. Introduction Hesperidin is 3′,5,7-trihydroxy-4′methoxy flavanone 7-o-β-rutinoside which is greatly found in citrus species and is the active constituent of tangerine peel (citrus reticulate anti-inflammatory effect by the inhibition of eicosanoids synthesis other biological activities including blood cholesterol lowering effect, hypotensive effect, protective agent against sepsis, antioxidant activity, and diuretics). It prevents poisoning caused by lead strontium and heavy metals. It is also used for diabetics and gastroesophageal reflux diseases. Hesperidin converted to hesperitn by intestinal microflora and subsequently absorbed from intestinal mucosa (see Figure 1). Figure 1: Chemical structure of Hesperidin. Diosmin is a synthetic drug (modified Hesperidin) of the flavonoid family. It is an oral phlebotropic drug used in the treatment of venous disease, that is, chronic venous insufficiently (CVI) and hemorrhoidal diseases. Diosmin aglycone is diosmetin; IUPAC name is 3′,5,7-trihydroxy-4 methoxy flavone 7-rutinosode. It might have potential in the treatment of neurodegenerative disease such as alzheimer’s diseases, and its anti-inflammatory and antiapoptotic activities have been demonstrated in neuronal