A Green Analytical Stability-Indicating RP-HPLC method for Simultaneous Determination of Anti-Hypercholesterolemic Agents and Their related Impurities

A straightforward, trustworthy and eco-friendly RP-HPLC-UV stability-indicating technique for separation and quantification of Bempedoic acid(BPA), Ezetimibe(EZM) and their impurities(BPA Imp-1, BPA Imp-2, EZM Imp-A, EZM Imp-B) was developed and validated in accordance with ICH Q2(R2) for accurate determination of BPA & EZM by minimizing the interference from impurities with impurity analysis simultaneously. Chromatographic separation was accomplished on an Inertsil ODS C18 Column (250×4.6mm, 5µm) using gradient elution mode with 0.1% TFA and Acetonitrile at a flow rate of 1.0 ml/min, with detection wavelength of 211nm and total run time is 40mins. Method reliability was assessed through key parameters such as specificity, linearity, accuracy, precision, sensitivity (LOD & LOQ) and robustness. The principles of green analytical chemistry were followed to minimize the usage of solvents. The results of linearity were consistent with R ² values above 0.999; percent recovery is in the range of 99–101%; RSD% values were below 2.0% for precision. LOD and LOQ values were found to be 0.03 and 0.1 µg/ml for impurities (BPA Imp-1, BPA Imp-2, EZM Imp-A, EZM Imp-B), 5.4 & 18µg/ml for BPAand 0.3 & 0.1µg/ml for EZM. By inducing degradation under extreme conditions, the stability-indicating property of the approach was explored.The approach was found to be effective in differentiating between BPA, EZM and their impurities showcasing the suitability of the method for routine quality control.The results of the devised technique confirm the reliability for quantification of analytes and impurities in their fixed dose combination. The AGREE and GAPI tools used to assess the greenness of the method showed a score of 0.72 and 85 showcasing reasonable balance among environmental sustainability and analytical performance, indicating the eco-friendliness of the developed method.