DEVELOPMENT AND VALIDATION OF A HPLC ANALYTICAL METHOD FOR DETERMINATION OF ELLAGIC ACID IN EPILOBIUM ANGUSTIFOLIUM EXTRACT

Development of quality assessment parameters for natural products is a prominence necessity to justify their acceptability and activity. Establishment of authentic and reliable analytical methods which profile the quantitative phytochemical composition of marker constituents in multicomponent composition like extract is a challenging task. A simple, rapid, precise, and reliable HPLC method was developed for the separation and estimation of ellagic acid from Epilobium angustifolium (Canadian willow herb) extract. The estimation was carried out using Sunfire C18 column, 5μ (4.6 × 250 mm); mobile phase consisting of 0.1% orthophosphoric acid and acetonitrile; the flow rate of 1 mL/min and ultraviolet detection at 280 nm with a properly resolved having run time of 35 min. The method was validated as a final verification of method development concerning precision, linearity, accuracy, ruggedness, and robustness. The correlation coefficient (r²) > 0.999, a method is considered to be linear as the correlation coefficient was found to be within acceptance criteria. The % RSD of peak area response due to ellagic acid in five replicate injections of standard solution was to be less than 2.0%, and system suitability parameters were passed. The % Average recovery of ellagic acid in Canadian willow Extract observed within the acceptance criterion of 98 – 102% indicates the accuracy of the method. The present validation proves that the HPLC-method is suitable for the determination of assay of ellagic acid from Canadian willow herb, extract at prescribed conditions.