Ch. Ramesh, D. Rama Devi, MNB. Srinivas, Nagaraju Rajana, S. Radha Krishna, K. Basavaiah
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Ch. Ramesh1,2*, D. Rama Devi3 , MNB. Srinivas1,2, Nagaraju Rajana1, S. Radha Krishna2, K. Basavaiah1*
1Department of Inorganic and Analytical Chemistry, Andhra University, Visakhapatnam - 530003, India.
2Laurus Labs Ltd., Visakhapatnam-531021, India.
3A.U. College of Pharmaceutical Sciences, Andhra University, Visakhapatnam, 530003, India.
Volume - 13,
Issue - 5,
Year - 2020
A stereospecific RP-HPLC method for the separation and estimation of S-isomer content in tenofovir disoproxil fumarate drug substance was developed and validated on a reverse-phase amylose derivativesed chiral column. The effect of organic modifiers, namely, methanol, acetonitrile and triethylamine in mobile phase was optimized as 0.1% triethyl amine in mixture of water, methanol and acetonitrile (10:75:15, v/v/v) to obtain the best enantiomeric separation. UV detection was performed at 260nm. The flow rate was kept at 0.8 mL/min and the column temperature was set at 25°C. The retention times of tenofovir and its S-isomer were observed to be 5.137 min and 8.768 min respectively. The linear regression analysis data for calibration plots showed a good linear relationship over a concentration range of 0.0005mg/mL – 0.01mg/mL for S-isomer. The values of the correlation coefficient were 0.999 for S-isomer. The limit of detection (LOD) was 0.0001 mg/mL and the limit of quantification (LOQ) was 0.0005mg/mL. The precision of S-isomer at LOQ level was evaluated through six replicate injections and the % RSD of the peak response achieved is 3.07. The percentage recoveries of S-isomer from tenofovir disoproxil fumarate were ranged from 96.4% to 102.2%. The proposed method was found to be accurate, precise and suitable for the separation and enantiomer excess determination of unwanted S-isomer in active pharmaceutical ingredients (API). The analytical results were supported by statistical parameters. The proposed method could be successfully applied to the enantiomeric purity analysis of tenofovir disoproxil fumarate in drug substance. This method was validated in as per ICH Q2 (R1) and USP validation of compendial methods <1225>.
Cite this article:
Ch. Ramesh, D. Rama Devi, MNB. Srinivas, Nagaraju Rajana, S. Radha Krishna, K. Basavaiah. Development and Validation of Reverse Phase HPLC Method for Enantiomer Excess Determination of Tenofovir Disoproxil Fumarate drug Substance. Asian J. Research Chem. 2020; 13(5):334-340. doi: 10.5958/0974-4150.2020.00064.4
Ch. Ramesh, D. Rama Devi, MNB. Srinivas, Nagaraju Rajana, S. Radha Krishna, K. Basavaiah. Development and Validation of Reverse Phase HPLC Method for Enantiomer Excess Determination of Tenofovir Disoproxil Fumarate drug Substance. Asian J. Research Chem. 2020; 13(5):334-340. doi: 10.5958/0974-4150.2020.00064.4 Available on: https://ajrconline.org/AbstractView.aspx?PID=2020-13-5-6
1. Drug bank, Tenofovir disoproxil fumarate, http://www.drugbank.ca/drugs/DB00300
2. Indian Pharmacopoeia, Government of India, Ministry of Health and Family Welfare, Published by The Indian Pharmacopoeia Commission, Ghaziabad, 2018, 2, 3327-3328.
3. Koen K.A. Van Rompay, Michael B. McChesney, Nancy L. Aguirre, Kimberli A. Schmidt, Norbert Bischofberger, Marta L. Marthas: Two Low Doses of Tenofovir Protect Newborn Macaques against Oral Simian Immunodeficiency Virus Infection. The Journal of Infectious Diseases, 2001, 184(4), 429-438.
4. Ryan D. Cooper, Natasha Wiebe, Nathaniel Smith, Philip Keiser, Saraladevi Naiker and Marcello Tonelli, Systematic review and Meta- analysis: Real safety of Tenofovir disoproxil fumarate in HIV-infected patients, Clinical Infections Diseases, 2010, 51(5), 496-505.
5. Joel E, Gallnt, Stanley Deresinki, Tenofovir disoproxil fumarate, Clinical Infectious Diseases, 2003, 37(7), 944-950.
6. Kathleen squires, Nicholaos C, Bellos and John J. Tooole, Tenofovir disoproxil fumarate in Nucleoside-Resistant HIV-1 Infection, A Randomized Trail, Ann Intern Med. 2003, 139, 313-320.
7. Keamey BP, Flherty JF, Shah J. Tenofovir disoproxil fumarate: Clinical Pharmacology and Pharmacokinetics. Clin Pharmacokinet 2004, 43, 595-612.
8. C. Brown, Chirality in Drug Design and Synthesis; Academic Press: London, UK, 1990.
9. Federsel, H.J. Chiral drug discovery and development from concept stage to market launch. In Comprehensive Medicinal Chemistry II; Elsevier: Amsterdam, The Netherlands, 2007.
10. Islam, M.R., Mahdi, J.G. and Bowen, I.D. Pharmacological Importance of Stereochemical Resolution of Enantiomeric Drugs. Drug-Safety 1997, 17, 149-165.
11. Borman, S. FDA issues flexible policy on chiral drugs. Chemical and Engineering News Archive 1992, 70, 24, 5 (News).
12. Stinson, S.C. In wake of new FDA guidelines, most drug firms are developing single enantiomers, spawning a “chirotechnology” industry. Chiral Drugs, Chemical and Engineering News Archive 1992, 70, 39, 46-79 (News).
13. BingyunLi, DonaldT.Haynie, Chiral Drug Separation, Encyclopedia of Chemical Processing, 2005.
14. Ozlem Coskun, Separation techniques: Chromatography, North Clin Istanb. 2016, 3(2), 156–160.
15. Zaheer Ahmed, Sathish Kumar Shetty, B. Gopinath, Manzoor Ahmed and B. K. Sridhar, Development and Validation of RP-HPLC Method for the Determination of Tenofovir Disoproxil in Bulk and in Pharmaceutical Formulation, Int. J. Chem. Sci., 2009, 7(4), 2447-2458.
16. Tripti Sharma, Neelam Mishra, A validated RP-HPLC method for estimation of tenofovir disoproxil fumarate in bulk and pharmaceutical formulation, Asian J Pharm Clin Res, 2012, 5(3), 108-110.
17. Shweta Havele and Sunil R. Dhaneshwar, Development and validation of a stability-indicating lc method for the determination of tenofovir disoproxil fumarate in pharmaceutical formulation, Songklanakarin J. Sci. Technol. 2012, 34 (6), 615-622.
18. Suresh babu bodempudi, Ravi chandra babu rupakula, Konda S Reddy, Development and validation of RP-chiral HPLC method for quantification of (s)-isomer in tenofovir disoproxil fumarate, Int J Curr Pharm Res, 2017, 19(6), 31-36.
19. Seshachalam U, Rajababu B, Haribabu B, Chandrasekhar KB. Enantiomeric separation of tenofovir on an achiral C18 column by HPLC using L-phenylalanine as a chiral mobile phase additive. J Liq Chromatogr Relat Technol 2008, 31, 410-20.
20. Yoshio Okamoto, Eiji Yashima. Polysaccharide Derivatives for Chromatographic Separation of Enantiomers. Angewandte Chemie, 1998, 37(8), 1020-1043.
21. E. Yashima, H. Iida and Y. Okamoto, Enantiomeric Differentiation by Synthetic Helical Polymers, Differentiation of Enantiomers I, 2013.
22. Karen D. Ward, Ardith D. Bravenec and Timothy J. Ward, Chiral separations by High Performance Liquid Chromatography, Encyclopedia of Analytical Chemistry, 2019.
23. International Conference on Harmonization of technical requirements for registration of pharmaceuticals for human use ICH harmonized tripartite guideline Validation of analytical procedures: text and methodology Q2 (R1), step 4 (2005).
24. General Information /USP Chapter <1225> Validation.