Analytical Method Validation of RP-HPLC Method for Simultaneous Estimation of Levonorgestrel and Ethinylestradiol from Combined Drug Product

摘要

The aim of recent study is to explore a rapid, sensitive and selective Reversed Phase HPLC method for simultaneous quantification of steroidal hormones Levonorgestrel and Ethinylestradiol from drug products. Chromatographic separation of Levonorgestrel and Ethinylestradiol were achieved by Kromasil C8 column (4.6 mm × 150 mm, 5 μm) at 4.8 and 3.9 min by using 60:40 volume of acetonitrile and deionized water mobile phase at a flow rate of 1.0 ml/min at wavelength of 247 nm and 310 nm respectively. The injection volume was 100 μL and the column temperature was maintained at 25°C. The method was validated according to USP category I requirements which includes specificity, accuracy, precision, intermediate precision, linearity and range and robustness. The standard calibration curve were found linear (r20.99) over the analytical range throughout the analysis day. Precision and intermediate precision was 3 % for Levonorgestrel and Ethinylestradiol at QC standard range. The method was found to be accurate in the range of 50 to 150% from QC standard. This method was also found robust in accordance with variation in the flow rate (± 0.5 ml/min.), organic phase ratio (± 10%), change column oven temperature (± 5°C), auto sampler temperature (5°C and 25°C) and changing column brand. The HPLC method was successfully applied to the analysis of dissolution samples of marketed Levonorgestrel and Ethinylesradiol combined tablets.