Analysis of ellagic acid in Punica granatum flower extract by high performance liquid chromatography.

摘要

Punica granatum is a shrub or tree, 8 to 10 meters high from the punicaceae family and grows wild in Iran. Flowers are regular, solitary or in fascicles at apices, with a size of 4 to 6 cm. Petals are lanceolate, wrinkled and brilliant orange-red. Punica granatum flower (PGFL), an important medicinal plant with wide medicinal properties, is frequently used in various traditional herbal preparations. The aim of this study was to determine ellagic acid in hydroalcoholic extract of PGFL, by high performance liquid chromatography (HPLC). Ellagic acid (EA), major bioactive phenol was selected as chemical marker of PGFL, and RP-HPLC method was developed for the quantification of EA using Inertsil C18 (250x4.6) mm, 5 micro column. The detection was carried out at wavelength 254 nm. The ellagic acid was satisfactorily resolved with retention time (RT) value about 38.2 minutes. The linear regression analysis data for the calibration plots showed good linear relationship with r2=0.9987. According to the International Conference on Harmonization (ICH) guidelines, the method was validated in terms of its linearity, specificity, recovery, precision, robustness and solution stability by following standard protocols. Statistical analysis of the data showed that the method is reproducible and selective for the determination of ellagic acid. The method was able to identify and quantify EA from complex mixtures of phytochemicals containing PGFL.