ANALYTICAL METHOD DEVELOPMENT AND METHOD VALIDATION FOR DETERMINATION ASSAY AND CONTENT UNIFORMITY OF LEVONORGESTREL BY REVERSED-PHASE HIGHPERFORMANCE LIQUID CHROMATOGRAPHY
Keywords:Method development, Method validation, Levonorgestrel, Reversed-phase high-performance liquid chromatography
Object: The main objective of the complete study is to develop a new method and also to validate the developed method for the determination of Assay and Content Uniformity of Levonorgestrel by reverse-phase high performance liquid chromatography (RP-HPLC).
Methods: RP-HPLC method was developed for simultaneous estimation of levonorgestrel using Hypersil ODS, 125 mm×4.6 mm×5 μm C8 column with a mixture of water, and acetonitrile solution with a ratio of 50:50 as a mobile phase at a flow rate of 1.3 mL/min with a detection of quantification wavelength of 242 nm. Method was selected after calculating system suitability and validated as per International Conference on Harmonization (ICH) guidelines.
Results: The developed analytical method parameters found within the limits as given in ICH and USP Guidelines and the total chromatographic analysis time per sample was 8 min with Levonorgestrel Eluting with retention time of 4.479, 4.479, and 4.467 min, respectively. The validated HPLC method was successfully applied for the determination of dissolution of levonorgestrel tablets.
Conclusion: The method is simple, precise, specific, and accurate. The newly developed method can be used for routine analysis of Levonorgestrel in tablet dosage form.
Ahuja S, Dong M. Handbook of Pharmaceutical Analysis by HPLC. 1st ed. Amsterdam: Elsevier; 2005.
Snyder LR, Kirkland JJ, Glajch JL. Practical HPLC Method Development. 2nd ed. Hoboken: John Wiley & Sons, Inc.; 1997.
Ravindra A, Hima P, Swamy KN, Kumar KV. Validated RP-HPLC method for simultaneous estimation of levonorgestrel and ethinylestradiol in combined dosage form. J Sci Innov Res 2013;2:642-50.
Basarabad CN, Westhoff CL, Pike MC, Nandkumar R, Cremers S. Estimating systemic exposure to levonorgestrel from an oral contraceptive. Contraception 2017;95:398-404.
Back DJ, Bates M, Breckenridge AM, Hall JM, MacIver M, Orme ML, et al. The pharmacokinetics of levonorgestrel and ethinylestradiol in women-studies with ovran and ovranette. Contraception 1981;23:229-39.
Fotherby K. Potency and pharmacokinetics of gestagens. Contraception 1990;41:533-50.
Fotherby K. Pharmacokinetics of gestagens: Some problems. Am J Obstet Gynecol 1990;163:323-8.
Seifeldeen EM, Etman MA, Aboul-Enein HY. Simultaneous determination of four hormonal compounds in oral contraceptive tablet formulations by high performance liquid chromatography. Taylor Francis Online 2016;39:1-24.
Dasari P, Veerareddy A, Bhoomireddy R, Chvsl K, Bethi M. Development and validation of stability indicating RP-HPLC method for the determination of impurity profile in gamendazole: Experimental male oral contraceptive. J Liq Chromatogr Relat Technol 2015;38:37-41.
Charde MS, Welankiwar AS, Kumar J. Method development by liquid chromatography with validation. Int J Pharm Chem 2014;4:57-61.
Gupta V, Deep A, Jain K, Gill NS, Gupta K. Development and validation of HPLC method-a review. Int Res Pharm Appl Sci 2012;2:17-25.
Bhagyasree T, Injeti N, Azhakesan A, Rao UM. A review on analytical method development and validation. Int J Pharm Res Anal 2014;4:444-8.
Chan CC, Leo YC, Lam H. Analytical Method Validation and Instrument Performance Verification. Vol. 1. USA: Wiley Interscience; 2004.
George N. Force degradation studies as an integral part of HPLC stability indicating assay method development. J Drug Deliv Technol 2010;10:1-4.
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