ABSTRACT:
This study aims to develop and validate RP-HPLC method for the estimation of ellagic acid in Acalypha indica extract. The isocratic RP-HPLC method was developed using a C18 column with a mobile phase consisting of methanol and 0.1% orthophosphoric acid (75:25 v/v), operated at a flow rate of 0.9 mL/min. A central composite design was employed for method optimization considering mobile phase composition and flow rate as critical process parameters and retention time and plate count were chosen as the critical analytical attributes. The method was validated for analytical parameters such as linearity, precision, accuracy, specificity, selectivity and the limits of detection (LOD) and quantification (LOQ). The optimized run was validated as per ICH guidelines. A strong linear relationship (R² = 0.9984) was established within the concentration range of 5–25 µg/mL. The method demonstrated excellent reproducibility and recovery evident by %RSD value which was less than 2. The LOD and LOQ were determined to be 0.141 µg/mL and 0.430 µg/mL, respectively. The method exhibited precise, accurate, robust and rugged characteristics as shown by % RSD values which were less than 2. The measured concentration of EA in Acalypha indica L. extracts were found to be 5.5034 µg/mL. The validated RP-HPLC method is sensitive, precise, accurate and robust, making it an effective tool for the reliable quantification of EA in Acalypha indica L. extracts. Hence, it can be concluded that developed method can be used for EA analysis in plant extract as part of quality standardization.