A RAPID STABILITY INDICATING HPLC-METHOD FOR DETERMINATION OF NORETHISTERONE ACETATE IN PLASMA, TABLETS AND IN A MIXTURE WITH OTHER STEROIDS
Objectives: This study reports about a simple, rapid and accurate, routine-HPLC method for quantification of Norethisterone acetate (NA) in bulk powder, tablets and plasma.
Methods: The chromatographic separation is carried out using isocratic binary mobile phase consisting of acetonitrile (ACN) and water (H2O) in the ratio of 55: 45 at the flow rate of 1 mLmin-1 and 40 Â°C. A diode array detector is used at 240 nm for detection.
Results: The elution time of NA is found to be 2.946Â±0.01 minutes. The method is validated for system suitability, linearity, precision, limits of detection and quantitation, specificity, stability and robustness. The robustness study is done for small changes in temperature, flow rate, the wavelength of detection, mobile phase content of ACN and injection volume. Stability tests are done through exposure of the analyte solution for four different stress conditions: Reflux with 1N hydrochloric acid (HCl), reflux with 1N sodium hydroxide (NaOH), reflux with 30% hydrogen peroxide (H2O2) and exposure to ultraviolet radiation (UV) radiation. The limits of detection and quantitation are 0.0625 and 0.125 ÂµgmL-1, respectively. The recovery value of this method is 101.90% and the reproducibility is within 2.16.
Conclusion: This is a rapid stability indicating HPLC method for determination of NA and can be applied for determination of NA in presence of other steroids as Levonorgestrel, Estradiol and Norethisterone.
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