Validation of Newly Developed Analytical Method for Standardization of Apigenin Using Rp-Hplc Method in Prepared Extract

摘要

In this study, a simple, precise and accurate analytical method was developed and validated for identification of apigenin using RP-HPLC method in prepared extract. Spectrophotometric determination was performed on a Perkin-Elmer UV-VIS Double Beam Spectrophotometer to know the maximum absorbance of the compounds. Chromatographic separation was performed using merck C18 analytical column (5 μm, 250 mm x 4.6 mm, i.d). Phosphate buffer at acidic pH and acetonitrile in the ratio of 30:70 v/v was considered to be suitable solvent system. The effluents were detected by means of UV detector at 268nm. The calibration curves were linear at a range of 10 - 50µg/ml with significant correlation coefficient of 0.9996. The retention time was found to be at 3.53min. The method was validated according to ICH guidelines and was found to contain the %RSD values below 2% which shows that the method was precise, specific and accurate.