DoE Approach: A validated Stability Indicating RP-HPLC Method Development for the Separation of Diasteromeric Analogs and Process Impurities of Carfilzomib

Abstract A novel reverse phase high performance liquid chromatographic (RP-HPLC) method was developed for the determination and separation of thirty diastereomers and six process related impurities of Carfilzomib (CFZ) active ingredient using YMC-Pack ODS-A (150 mm x 4.6 mm, 3µm) column and mobile phase comprising of pH 5.5 potassium di-hydrogen phosphate buffer, acetonitrile and methanol. The flow rate and detection wavelength were 0.9mL min−1 and 220 nm respectively. Design of Experiment (DOE) approach was strategically designed to optimize the critical chromatographic parameters like column temperature, flow rate and composition of methanol in the mobile phase to achieve the separation of diastereomers and process impurities. The developed method was validated for linearity, range, precision, accuracy, specificity, selectivity and intermediate precision.