Development and Validation of method for the determination of related substances of Norethindrone in Norethindrone Tablets and Degradation studies

A gradient, reversed-phase liquid chromatographic (RP-LC) method was developed for the quantitative determination of Norethindrone in Norethindrone tablets, used to treat drug for contraception and hormone replacement therapy. The gradient LC method employs solutions A and B as mobile phase. The solution A contains a mixture Milli Q water, acetonitrile and tetrahydrofuran in the ratio of 40:30:30 v/v/v and solution B contains a mixture Milli Q water, acetonitrile and tetrahydrofuran in the ratio of 40:30:30 v/v/v. The Chromatography was performed on a Supelco Ascentis Express C-18, 4.6 x 150mm, 2.7μ and detector of UV at 256 nm.0.9 mL/min as a Flow rate, 100 μL as an Injection volume. The chromatogram of Norethindrone its impurities namely NE-H (Impurity-H), NE-B (Impurity-B), NE-C (Impurity-C) and NE-D (Impurity-D) were found. Accuracy satisfactory by % recovery obtained in the range of 94.9 – 105.9, the linearity results for Norethindrone and impurities in the specified concentration Calibration curves were linear with a coefficient of variation (r) not less than 0.99. An accelerated degradation study on Norethindrone Tablets as following conditions Hydrolytic and Oxidative degradation, Thermal degradation, Humidity degradation, Photolytic degradation, Forced degradation studies (Acid, Alkali , Peroxide Thermal, Humidity and Photolytic ).The proposed method was found to be specificity, linearity, and precision, intermediate precision, and accuracy, stability in analytical solution and robustness. The validation was performed according to the current requirements as laid down in the ICH guidelines.