Abstract

RP-HPLC method has been proposed on phenomenex BDS C18 column (250 × 4.6 mm, 5μ) using the mixture of methanol: phospahte buffer, 10 mM (pH adjusted to 2.0 with 0.1% O-phosphoric acid) in the ratio of 84:16 v/v as mobile phase. The flow rate was maintained at 1mL/min and the analytes were monitored at 220 nm. Ethinyl Estradiol and Norethindrone Acetate eluted at 3.5 and 4.7 min respectively with resolution of 4.17 min. The peaks obtained were symmetrical with tailing factor less than 1.5 and theoretical plates more than 2000. The developed method was validated in accordance with ICH guidelines and the results of all parameters were found within acceptable limits. The method was found linear in the concentration range of 5-1000 μg/mL and relative standard deviation of the precision study was found about 2.0%. The assay results was found 102.94 (±2.01) and 105.37 (±1.94) % with mean % Recovery 106.30 and 95.45 for Ethinyl Estradiol and Norethindrone Acetate respectively. The developed method has been found suitable for routine analysis of Ethinyl Estradiol and Norethindrone Acetate in mixture and formulation.