Development, Validation and Forced Degradation Study of Emtricitabine and Tenofovir Alafenamide in its Pharmaceutical Dosage Form Using RP-HPLC
Journal of Pharmaceutical Research International,
Page 37-46
DOI:
10.9734/jpri/2021/v33i43A32462
Abstract
Aims: The present research was aimed to develop and validate a reverse phase high performance liquid chromatographic (RP-HPLC) method for the quantification of Emtricitabine (EMT) and Tenofovir Alafenamide (TEN) in combination.
Methodology: Separation was achieved under optimized chromatographic condition on an Inertsil C18, 250 x 4.6 mm, 5μm column. Various composition of mobile phase was tried. Separation of EMT and TEN was started with Methanol: Buffer and Methanol finally using solvent system of Buffer (pH 3.5) and Methanol in ratio of (30:70) and flow rate adjust at 1.0 ml/min was used as solvent system, the detection was carried out at 262nm using Shimazdu UV-visible detector. The mobile phase run time for the developed analytical method was 10 minutes.
Results: The standard curve was found linear in the concentration range of 20-60 μg/ ml (r2- 0.9994) and 2.5-7.5 μg/ ml (r2-0.9992) for EMT and TEN respectively. The %RSD was found to be 0.80-0.95% and 0.63-1.09 for EMT and TEN respectively. Percentage (%) recoveries for EMT and TEN to be in range of 100%-100.6% and 99.32%-100.83% respectively. The limit of detection and the limit of quantification were found to be 4.80 μg/ ml and 14.7 μg/ ml respectively for EMT and 0.11 μg/ ml and 0.33μg/ ml respectively for TEN. Results of forced degradation study showed EMT degradation in acid and base medium while TEN was showed degradation in oxidative stress. The proposed developed RP-HPLC method was validated statistically and the values were found to be within the acceptable limits.
Conclusion: In conclusion, the developed RP-HPLC method was found to be simple, specific, and rugged for simultaneous estimation of EMT and TEN. Validation results of method was found within the acceptable limits. Hence it can be used for analysis of EMT and TEN.
Keywords:
- Degradation
- RP-HPLC
- EMT
- TEN
- validation
How to Cite
References
Introduction of Tenofovir Alfenamide Hemifumarate; 2018.
Available:https://www.sciencedirect.com/topics/neuroscience/tenofovir-alafenamide-fumarate
Valli purnima B, Santha kumari M, Ramu G, Vijaya T, Reddy B, Ramachandran D. Simultaneous determination of emtricitabine and tenofovir desoproxil fumerate in truvada by derivative spectrophotometry. Asian Journal of Chemistry. 2017;29(5):1069-1075.
Joshi M, Nikalje A, Baig MS, Dehgan MHG. HPTLC method for the simultaneous estimation of emtricitabine and tenofovir in tablet dosage form. Indian Journal of Pharmaceutical Sciences. 2009;71(1):95-7
Dutta JK, Rajan G. Reverse-phase high-performance liquid chromatography method development and validation for simultaneous estimation and forced degradation studies of emtricitabine, rilpivirine, and tenofovir alafenamide in solid dosage form. Asian Journal of Pharmaceutical and Clinical Research. 2019;12(1):112-116.
Singh VSG, Divakar TE. A novel stability indicating RP-HPLC method development and validation for simultaneous estimation of bictegravir, emtricitabine and tenofovir in pure and fixed dose combination. Asian J. Org. Med. Chem. 2019;4(1):7-13.
Appala Raju VVSS, Ramana MV, Shaym T, Rajnikanth KN, Kumar K, Appla raju N. Bioanalytical method development and validation for simultaneous determination of bictegravir, tenofovir and emtricitabine in human plasma by LC-MS/MS. World Journal Of Pharmacy And Pharmaceutical Sciences. 2018;7(12):441-463.
Valko K, Snyder LR, Glajch J. Retention in reversed-phase liquid chromatography as a function of mobile phase composition.‖ J. Chromatogr. A. 1993;656(1-2):501–20.
Neue UD. HPLC columns: Theory, technology, and practice‖, John Wiley & Sons, New York. 1997;926-936.
Petersson P. RPLC column classification and the development of a column selection tool‖, ACD/Labs European Users Meeting, Obernai, France. 2003;23-24.
Huber JFK, Van der Linden R, Ecker E, Oreans M. Column switching in high pressure liquid chromatography.‖ J Chromatogr. 1973;2:267-71.
Yun KS, Zhu C and Parcher JF. Theoretical relationships between the void volume, mobile phase volume, retention volume, adsorption, and Gibbs free energy in chromatographic processes.‖ Anal. Chem. 1995;4:613–9.
Heinisch S, Rocca JL. Effect of mobile phase composition, pH and buffer type on the retention of ionizable compounds in reversed-phase liquid chromatography: Application to method development.‖ J. Chromatogr. A. 2004;1048(2):183–93.
Gritti F, Guiochon G. Role of the buffer in retention and adsorption mechanism of ionic species in reversed-phase liquid chromatography: I. Analytical and overloaded band profiles on Kromasil-C18.‖ J. Chromatogr. A. 2004;1038(1-2):53–66.
Bosch E, Espinosa S, Roses M. Retention of ionizable compounds on high performance liquid chromatography: III. Variation of pKa values of acids and pH values of buffers in acetonitrile–water mobile phases.‖ J Chromatogr A.1998;824(2);137–46.
Bosch E, Bou P, Allemann H, Roses M. Retention of ionizable compounds on HPLC: pH scale in methanol–water and the pKa and pH values of buffers. ‖ Anal. Chem. 1996;20:3651–7.
ICH guideline Q1A (R2). Stability testing of New Drug Substances and Products.
ICH guideline Q1B. Stability testing: Photostability testing of new drug substances and products.
ICH Q3B(R2). Impurities in New Drug Products
Blessy M, Patel RD, Prajapati PN, Agrawal YK. Development of forced degradation and stability indicating studies of drugs: A review, Journal of Pharmaceutical Analysis, 2013;09:003.
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