A Novel Validated stability indicating QBD based RS method by HPLC for the estimation of impurities of Norethindrone in Norethindrone acetate and their degradation as per ICH Q2 Guideline

A novel and efficient stability indicating reverse phase High performance liquid chromatography with diode array detector (RP-HPLC-DAD) RS analytical method has been developed, optimized and validated for the determination of degradation products and process impurities of Norethindrone in Norethindrone tablets: Nor androstenedione (Imp-A), Norethindrone enolether (Imp-B), Delta-5(6)Norethindrone (Imp-C), Delta-5(10)Norethindrone (Imp-D). The impurities elution and separation were established by optimized gradient chromatographic parameter using Inertial ODS-3V (150 mm x 4.6 mm) 5μ analytical column, water and acetonitrile as mobile phase, with run time 55 minutes, diode array detector (DAD) was set up at 210 nm and 240 nm wavelength for the simultaneous analysis of all impurities. The developed HPLC method was validated as per the International Conference on Harmonization (ICH) guidelines. The validation parameters used are Specificity, linearity, accuracy, precision, intermediate precision, and robustness. Limit of detection (LOD) and limit of quantification (LOQ) were also obtained for all impurities. Norethindrone samples and placebo were subjected to stress conditions of hydrolysis (acid and base), oxidative, and thermal stress degradation. Sample and standard solutions stability was also performed. The proposed validated method was successfully used for the quantitative analysis of impurities and degradation products of Norethindrone tablets. The method was found to be suitable for the stability, release, in process and Quality control analysis. The developed method is stability indicating, specific, selective, simple, precise, cost effective, linear and robust