Stability Indicating HPTLC for Quantitative Estimation of Dolutegravir, Lamivudine, and Tenofovir in Bulk and Dosage Form
Pharmaceutical sciences- Pharmaceutical analysis
DOI:
https://doi.org/10.22376/ijlpr.2023.13.6.P200-P216Keywords:
Dolutegravir, HPTLC, Lamivudine, Stability-indicating, Tenofovir, ValidationAbstract
Pharmaceutical companies are manufacturing multiple drug formulations to meet the market demand and treatunresponsive patients to single-drug formulations. Hence, an integral aim of the present study was to investigate the completedegradation behavior of the drug. It was done through a systematic investigation involving forced decomposition of the drug undervarious stress conditions. The study's main objective is to develop a Simple, accurate, and stability-indicating high-performance thinlayer chromatographic method for densitometric determination of Dolutegravir, Lamivudine, and Tenofovir in bulk and dosage form.The method development used TLC aluminum plates precoated with silica gel 60F 254 – (Merck, Germany) as the stationary phase.The solvent system consisting of Chloroform, Toluene, methanol, and ammonia (7:2:1:0.2v/v/v/v) was used for the development.Densitometric measurement of Dolutegravir, Lamivudine, and Tenofovir was performed at 260nm. The system gave very wellresolved spots for Dolutegravir, Lamivudine, and Tenofovir at Rf values of 0.29, 0.48, and 0.65, respectively. The calibration curve ofthe drug 400 -3200 ng/spot for DTG, 500 to 4000 ng/spot for LMV, and concentration range 1000-8000 ng/spot for TDF wasobserved. The correlation coefficient was 0.9995 for DTG, 0.9986 for LMV, and 0.9992 for TAF. The regression line equation forDolutegravir, Lamivudine, and Tenofovir was found to be y = 1.0427x + 792.51, y = 0.8791x, + 1556.3 and y = 1.0217x + 99.194,respectively. The %RSD less than two indicates the method is precise and accurate. Stability testing aimsStability testing aims to provehow the drug quality varies with time under the influence of various natural factors such as humidity, light, and temperature, enablingstorage conditions, retest periods, and determining the drug's shelf life. The degradation products were well resolved from the puredrug with significantly different retention factors in HPTLC. The method was proved to be a stability-indicating method and can beused in practice for bulk and dosage forms and to evaluate the shelf life.
References
Cottrell ML, Hadzic T, Kashuba ADM. Clinical pharmacokinetic, pharmacy dynamic and drug-interaction profile of the integrase inhibitor dolutegravir. Clin Pharmacokinet. 2013;52(11):981-94. doi: 10.1007/s40262-013-0093-2, PMID 23824675.
Coates JA, Cammack N, Jenkinson HJ, Jowett AJ, Jowett MI, Pearson BA et al.. (-)-2’-deoxy-3’-thiacytidine is a potent, highly selective inhibitor of human immunodeficiency virus type 1 and type 2 replication in vitro. Antimicrob Agents Chemother. 1992;36(4):733-9. doi: 10.1128/AAC.36.4.733, PMID 1380229.
The Merck. Index. 13th ed. Whitehouse Station, NJ: Merck and Company Inc; 2001. p. 1631-32.
Martindale. The complete drug reference. 33rd ed. Vol. 620. London: Pharmaceutical Press; 2002. p. 642.
Asnani AJ, Mohurle SM, Pratyush K. Development and validation of UV spectrophotometric method for estimation of lamivudine and tenofovir disoproxil fumarate in combined dosage form using quality by design approach. Int J Pharm Sci Res. 2021;12(6):3306-15. doi: 10.13040/Ijpsr.0975-8232.12(6).3306-15.
Bhavyasri K, Divya E. Method Development, validation and forced degradation studies of dolutegravir, an antiretroviral drug using UV-visible spectroscopy. Int J PharmSci Scient Res. 2019;5(6):69-74.
Dhanwate SS, Pachauri AD, Ghode PD, Khandelwal KR. Development and validation of an analytical method for estimating lamivudine and dolutegravir sodium in the dosage form. J Pharm Sci Res. 2019;11(8):2886-90.
A SA, H BC, J SS. Application of UV-spectrophotometric methods for estimation of tenofovir disoproxil fumarate in tablets. Pak J Pharm Sci. 2009;22(1):27-9. PMID 19168416.
Vidyadhara S, Sasidhar RLC, Rao BV, Kumari PR. Simultaneous UV spectrophotometric method for the determination of tenofovir, efavirenz, and lamivudine in Bulk and combined dosage form. Asian Jour Pharmac Anal. 2016;6(4):253-8. doi: 10.5958/2231-5675.2016.00037.5.
Dulange V, Gajeli GB. Development and validation of UV spectroscopy method for the estimation of Dolutegravir in bulk and pharmaceutical dosage form. Asian J Pharm Anal. 2021;11(3):188-90. doi: 10.52711/2231-5675.2021.00032.
Sriveni T, Naveen V, Rupa VS, Renuka A, Porika S, Haque MA et al. development and validation of Dolutegravir in bulk and formulation: an antiretroviral drug usingUV-spectroscopy. IJPQA.2021;12(1):57-60.
Masthanamma SK, Sridhar TA, Saidulu P. A Novel UV-spectrophotometric Method development and validation of Dolutegravir in bulk and its laboratory synthetic mixture by using 8 m urea as hydrotropic solubilizing agent. Int J Pharm Sci Drug Res. 2015;7(4):370-5.
Bhadauria RS, Gupta RK. Development and validation of UV spectrophotometric method for simultaneous estimation of lamivudine and tenofovir Disproxil fumarate in the combined dosage form. JDDT. 2020;9(4-s):1156-9.
U H, Akm P. Development and validation of stability indicating simultaneous UV spectrophotometric method for determination of emtricitabine, tenofovir disoproxil fumarate, cobicistat, and elvitegravir in pure and pharmaceutical dosage form. Asian J Pharm Clin Res. 2018;11(4):177-84. doi: 10.22159/Ajpcr.2018.V11i4.22897.
Bhirud CH, Hiremath SN. Development of validated stability-indicating simultaneous estimation of tenofovir disoproxil fumarate and emtricitabine in tablets by HPTLC. J Pharm Res. 2013;7(2):157-61 https:. doi: 10.1016/j .jopr.2013 .02.019.
Bizimana T, Kayumba PC, Védaste K, Egide K, Kaale E. Development of high performance thin layer chromatography for simultaneous analysis of lamivudine and tenofovir disoproxil fumarate. RWJour. 2016;3(1):6-10. doi: 10.4314/rj.v3i1.1F.
Bhavar GB, Pekamwar SS, Aher KB, Thorat RS, Chaudhari SR. High-performance liquid chromatographic and high-performance thin-layer chromatographic method for quantitative estimation of dolutegravir sodium in bulk drug and pharmaceutical dosage form. Sci Pharm. 2016;84(2):305-20. doi: 10.3797/Scipharm.1507-09, PMID 27222606.
Dhanve A, Rao JR, Dhale C. Development and validation of stability indicating HPTLC method for simultaneous estimation of lamivudine and dolutegravir sodium in bulk and pharmaceutical dosage formulation. Int J Pharm Sci Res. 2018;9(11):4701-4708doi. doi: 10.13040/ijpsr.0975-8232.9(11).4701-08.
Rao NM, Sankar DG. Development and validation of stability-indicating HPLC method for simultaneous determination of lamivudine, tenofovir, and Dolutegravir in bulk and their tablet dosage form. Future J Pharm Sci. 2015;1(2):73-7: https:. doi: 10.1016/j.fjps.2015.11.002.
M V, Jyothi N S. Development of validation and stability indicating method of anti-HIV Dolutegravir drug and its related impurities by using RP-HPLC. J Chromatogr Sep Tech. 2020;11(1):426. doi: 10.35248/2157-7064.20.11.426.
Patel K, Shah U, Patel D, Patel JK. Patel TB.development and validation of RP- HPLC stability indicating method for simultaneous estimation of Dolutegravir and Lamivudine in bulk and pharmaceutical dosage form. J Pharm Res Int. 2021;33(60B):570-84. doi: 10.9734/jpri/2021/v 33i60B 34656.
Anandakumar K, Kamarajanb K, Vetrichelvan TA. Validated RP-HPLC method for simultaneous estimation of lamivudine and tenofovir disoproxil fumarate in pure and the tablet dosage form. Eurasian J Anal Chem. 2012;7:56-66.
Purnima BV, Reddy TVB, Rao YS, Ramu G. Stability indicating RP-UPLC method for assay of emtricitabine and tenofovir disoproxil fumarate in bulk and dosage forms. Am J Anal Chem. 2015;6:807-21. doi: 10.4236/ajac.2015.610077.
Kumar KA, Nagpal KL, Kumar J. Development and validation of an LC-MS/MS assay for bioequivalence study of dolutegravir, emtricitabine, tenofovir alafenamide in human plasma. Int J Res Pharm Life Sci. 2017;5(2):40-53.
Babu C, Devanna N, Reddy KVN. Validated gradient stability indicating RP-HPLC method for the simultaneous quantification of 11 related substances in the combined dosage forms of lamivudine and tenofovir disopeoxil fumarate. Int J Appl Pharm. 2017;9(4):61-8. doi: 10.22159/ijap.2017v9i4.19001.
Godela R, Kammari V, Gummadi S, Beda D. Concurrent estimation of lamivudine, tenofovir disoproxil fumarate, and efavirenz in blended mixture and triple combination tablet formulation by a new stability indicating RP-HPLC method. Futur J Pharm Sci. 2021;7(1):94. doi: 10.1186/s43094-021-00249-9.
R S, T S, D G, M T. Analytical method development and validation of stability indicating assay method of analysis for dolutegravir/lamivudine/ tenofovir disoproxil fumarate tablets using high-performance liquid chromatography. Res J Pharm Technol. 2021;14(5):2434-9. doi: 10.52711/0974-360X.2021.00428.
Dubbaka A, Sireesha D, Bakshi V. Analytical method development and validation for the simultaneous estimation of lamivudine and tenofovir disoproxil fumarate by RP-HPLC method. MOJ Proteom Bioinform. 2016;4(5):306-9. doi: 10.15406/mojpb.2016.04.00134.
Dubey S, Duggirala M. Simultaneous estimation of lamivudine, abacavir, and dolutegravir by UPLC method. Int J Appl Pharm. 2018;10(1):46-52. doi: 10.22159/Ijap.2018v10i1.21156.
Rathod SM, Patel PU. Development and validation of RP – HPLC method for estimation of lamivudine and dolutegravir sodium in synthetic mixture. Res J Pharm Technol. 2020;13(6):2864-8. doi: 10.5958/0974-360X.2020.00510.7.
Sykes C, Blake K, White N, Schauer AP, Guzman BB, Cottrell ML et al. Development and validation of an LC-MS/MS assay for the quantification of dolutegravir extracted from human hair. Anal Bioanal Chem. 2018;410(29):7773-81. doi: 10.1007/s00216-018-1394-y, PMID 30280227.
Srivastava B, Sheeja VK, Haribabu Y, Ebin CJ. Method Development and validation of simultaneous estimation of Emtricitabine and Tenofovir Alafenamide in bulk and tablet dosage form using LC-MS/MS. Res J Pharm Technol. 2021;14(5):2493-6. doi: 10.52711/0974-360X.2021.00439.
Wang X, Penchala SD, Amara A, Else L, McClure M, Boffito M. A Validated method for quantification of Dolutegravir using ultra-performance liquid chromatography coupled with UV detection. Ther Drug Monit. 2016;38(3):327-31. doi: 10.1097/FTD.0000000000000286. PMID 26859200.
Madeesh SK, Ismail Y, Gunasekaran V. Development and validation for simultaneous estimation of lamivudine, tenofovir and efavirenz by UPLC. Int J Pharm. 2012;2:656-60.
Sonia K, Bhavyashree B. Lakshmi KS, HPTLC method development and validation: an overview. J Pharm Sci Res. 2017;9:652-7.
Srivastava MM. An overview of HPTLC: A modern analytical technique with excellent potential for automation, optimization, hyphenation, and multidimensional applications. In: Srivastava MM, editor. High-performance thin layer chromatography (HPTLC); 2010. p. 3-24. doi: 10.1007/978-3-642-14025-9_1.
Weins C, Hauck HE. Advances and developments in thin layer chromatography. LC-GC Int. 1996;14:455-71.
Reich E, Schibli A, Debatt A. Validation of high-performance thin-layer chromatographic methods for the identification of botanicals in a cGMP environment. J AOAC Int. 2008;91(1):13-20. doi: 10.1093/jaoac/91.1.13, PMID 18376581.
Swartz M. Chromatography: method validation. In: Wilson ID, Poole C, editors. Handbook of methods and instrumentation in separation science. CA: Academic Press; 2009. p. 614-20.
Koll K, Reich E, Blatter A, Veit M. Validation of standardized high-performance thin-layer chromatographic methods for quality control and stability testing of herbals. J AOAC Int. 2003;86(5):909-15. doi: 10.1093/jaoac/86.5.909, PMID 14632390.
ICH Guideline Q2. Validation of analytical procedures: text and methodology. Available from: http://www.ich.org/LOB/media/MEDIA417.pdf. Vol. R1; 2005.
Kats R. Forced degradation studies: regulatory considerations and implementation. Biopharm Int. 2005;18:1-7.
Reynolds WD. Forced degradation of pharmaceuticals. Am Pharm Rev. 2004;7(3):56-61.
Mangamma K, Rohini P, Manvitha GDP, Chandra UR, Rohini P. Stability indicating method development and validation for the simultaneous estimation of dolutegravir, lamivudine and tenofovir disoproxil fumarate by RP-HPLC. Int J Pharm Res Appl. 2020;5(2):16-23. doi: 10.35629/7781-05021623.
International Conference on Harmonization (ICH) of Technical Requirements for the Registration of Pharmaceuticals for Human Use, Validation of Analytical Procedures, ICH-Q2A, Geneva 1995.
Center for Drug Evaluation and Research, US Food and Drug Administration. Reviewer guidance and validation of chromatographic methods. Rockville, MD: FDA; 1994.
International Conference on Harmonization (ICH) of Technical Requirements for the Registration of Pharmaceuticals for Human Use, Validation of Analytical Procedures: Methodology, ICH Q2B, Geneva 1996.
Bakshi M, Singh S. Development of validated stability-indicating assay methods critical review. J Pharm Biomed Anal. 2002;28(6):1011-40. doi: 10.1016/s0731-7085(02)00047-x, PMID 12049968.
Gu Y, Zeng B, Sherman J. Development of Quantitative HPTLC Methods for dolutegravir, lamivudine, and tenofovir Disproxil fumarate in a combination pharmaceutical product using a model process published earlier for transfer of Minilab TLC screening methods to HPTLC-Densitometry. Acta Chromatographica. 2019;32(3):199-202. doi: 10.1556/1326.2019.00689.
Reynolds D, Facchine K. Available guidance and best practices for conducting forced degradation studies. Pharm Technol. 2002;26(3):48-56.
Chandramore K, Sonawane S. Overview on Development and Validation of Force degradation studies with Stability Indicating Methods. Biosci Biotechnol Res Asia. 2022;19(4):881-94. doi: 10.13005/bbra/3039.
Said NS, Nasr ZA, Abdel-Razeq SA. Assessment of the greenness of new stability indicating micellar UPLC and HPTLC methods for determination of tenofoviralafenamide in dosage forms. J Chromatogr Sci. 2021;59(10):909-22. doi: 10.1093/chromsci/bmaa143, PMID 33529317.
Alsante KM, Ando A, Brown R, Ensing J, Hatajik TD, Kong W et al. The role of degradant profiling in active pharmaceutical ingredients and drug products. Adv Drug Deliv Rev. 2007;59(1):29-37. doi: 10.1016/j.addr.2006.10.006, PMID 17187892.
Blessy M, Patel RD, Prajapati PN, Agrawal YK. Development of forced degradation and stability indicating studies of drugs—a review. J Pharm Anal. 2014;4(3):159-65. doi: 10.1016/j.jpha.2013.09.003, PMID 29403878.
Sutar SV, Yeligar VC, Patil SS. A review: stability indicating forced degradation studies. Res J Pharm Technol. 2019;12(2):885-90. doi: 10.5958/0974-360X.2019.00152.5.
Venkataraman S, Manasa M. Forced degradation studies: regulatory guidance, characterization of drugs, and their degradation products. Drug Invent Today. 2018;10(2):137.
Iram F, Iram H, Iqbal A, Husain A. Forced degradation studies. J Anal Pharm Res .2016. 2016;00073:3(6). doi: 10.15406/japlr.2016.03.00073.
Published
How to Cite
Issue
Section
Copyright (c) 2023 Charushila Bhangale, Shivanand Hiremath
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
