Validated HPLC Method for Quantitative Analysis of Gallic Acid and Rutin in Leaves of Moringa Oleifera Grown in Egypt

Moringa oleifera is one of miracle tree and great interested medicinal plant which belongs to Moringaceae. It is widely used as a nutritive herb and possesses valuable pharmacological activities. In our study a rapid, sensitive and accurate HPLC method for qualitative and quantitative determination of gallic acid and rutin in two different solvent methanol and ethanol, extracts of Moringa oleifera leaves cultivated in Egypt was investigated. The separation was performed and validated by isocratic method utilizing C18 column with a mobile phase 10% (v/v) acetonitrile and 1.0% aqueous acetic acid mixture for gallic acid and 60% (v/v) methanol and 1.0% aqueous acetic acid mixture for rutin. Those bioactive components, gallic acid and rutin, were detected at 280 and 335 nm, respectively. The calibration graph was found to be linear in the concentration ranges 5.0-40.0 and 5.0-60.0 µg/mL for gallic acid and rutin, respectively. The higher concentrations of gallic acid were observed in the 50% methanolic and ethanolic extracts, while the higher concentrations of rutin were observed in the 100% methanolic and ethanolic extracts. Moreover, a successfully spectrophotometric quantification of the total phenolics and flavonoids content in the plant extract was achieved, where the equivalent amount to gallic acid in methanolic and ethanolic extracts and the equivalent amount to rutin in methanolic and ethanolic extracts were calculated.