• V. L. N. BALAJI GUPTA TIRUVEEDHI Department of Basic Science, Vishnu Institute of Technology (A), Bhimavaram, Andhra Pradesh, India
  • VENKATESWARA RAO BATTULA Department of Engineering Chemistry, Andhra University, Visakhapatnam, Andhra Pradesh, India
  • KISHORE BABU BONIGE Department of Engineering Chemistry, Andhra University, Visakhapatnam, Andhra Pradesh, India




Doravirine, Tenofovir disoproxil fumarate, Lamivudine, Stability, RP-HPLC


Objective: In this study, a RP-HPLC (stability-indicating) based assay method for the estimation of doravirine (DRV), tenofovir disoproxil fumarate (TFF) and lamivudine (LMV) simultaneously in the tablets was described.

Methods: The simultaneous analysis of DRV, TFF and LMV was done with HPLC system (Agilent 1100 series) and Luna Phenomenex C18 (250 mm × 4.6 mm × 5 μ) column with isocratic mobile phase (35% volume ratio of methanol and 65% volume ratio of 20 mmol ammonium formate, pH 5). Validation of assay method was done on sensitivity, linearity, accuracy, selectivity, precision, robustness and specificity.

Results: The calibration curves were linear through the range of 25-200 µg/ml for DRV and 75-600 µg/ml for TFF and LMV. The percent relative standard deviation for intraday variation/precision, interday variation/precision, intermediate precision/ruggedness and robustness were lower than 2%. The recovery of LMV (99.09-99.76%), TFF (99.10-99.41%) and DRV (98.65-99.28%) confirmed the good accuracy. The stability of LMV, TFF and DRV in 0.1N NaOH, 3% peroxide, 0.1 N HCl, UV light and dry heat of 60 °C was determined.

Conclusion: The results have allowed the method to be implemented in the tablets to quantify DRV, TFF, and LMV.


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Rainer S. German advisory committee blood (Arbeitskreis Blut), subgroup ‘assessment of pathogens transmissible by blood’. Human immunodeficiency virus (HIV). Transfus Med Hemother 2016;43:203-22.

Nyamweya S, Hegedus A, Jaye A, Rowland Jones S, Flanagan KL, Macallan DC. Comparing HIV-1 and HIV-2 infection: lessons for viral immunopathogenesis. Rev Med Virol 2013;23:221-40.

Global HIV and AIDS statistics-2020 fact sheet, unaids.org. Available from: https://www.unaids.org/en/resources/fact-sheet [Last assessed on 05 Aug 2020]

Global Statistics, Hiv gov. Available from: https://www.hiv.gov/hiv-basics/overview/data-and-trends/global-statistics [Last assessed on 10 Aug 2020]

Drug and Device News. P and T 2018;43:588-98.

Pham HT, Xiao MA, Principe MA, Wong A, Mesplède T. Pharmaceutical, clinical, and resistance information on doravirine, a novel non-nucleoside reverse transcriptase inhibitor for the treatment of HIV-1 infection. Drugs Context 2020;9:11-4.

Delstrigo 100 mg/300 mg/300 mg film-coated tablets. Available from: https://www.medicines.org.uk/emc/product/9694/smpc [Last accessed on 05 Jun 2020]

Boyle A, Moss CE, Marzolini C, Khoo S. Clinical pharmacodynamics, pharmacokinetics, and drug interaction profile of doravirine. Clin Pharmacokinet 2019;58:1553-65.

Colombier MA, Molina JM. Doravirine: a review. Curr Opin HIV AIDS 2018;13:308-14.

Deeks ED. Doravirine: first global approval. Drugs 2018;78:1643-50.

Kearney BP, Flaherty JF, Shah J. Tenofovir disoproxil fumarate: clinical pharmacology and pharmacokinetics. Clin Pharmacokinet 2004;43:595-612.

Lyseng Williamson KA, Reynolds NA, Plosker GL. Tenofovir disoproxil fumarate: a review of its use in the management of HIV infection. Drugs 2005;65:413-32.

Quercia R, Perno CF, Koteff J, Moore K, McCoig C, St Clair M, et al. Twenty-five years of lamivudine: current and future use for the treatment of HIV-1 infection. J Acquir Immune Defic Syndr 2018;78:125–35.

Deepali G, Elvis M. UV spectrophotometric method for assay of the anti-retroviral agent lamivudine in active pharmaceutical ingredient and in its tablet formulation. J Young Pharm 2010;2:417-9.

Sonar KV, Sapkale P, Jadhav A, Deshmukh T, Patil S, Murkute P. Development and validation of UV spectroscopic method for estimation of lamivudine in tablet dosage form. Int J Curr Pharm Res 2017;9:86-9.

Vikram Singh A, Nath LK, Pani NR. Development and validation of analytical method for the estimation of lamivudine in rabbit plasma. J Pharm Anal 2011;1:251-7.

Kano EK, dos Reis Serra CH, Koono EE, Andrade SS, Porta V. Determination of lamivudine in human plasma by HPLC and its use in bioequivalence studies. Int J Pharm 2005;297:73-9.

Bakshi AN, Neelam T. Quantification and impurity profiling of anti-hiv drugs by HPLC MS/MS method. JCR 2020;7:4109-16.

Alekhya K, Srinivasan MS, Subramani S, Surya R, Vijey AM. Simultaneous evaluation of abacavir sulfate as well as lamivudine in medical formulations by gradient reversed-phase high-performance liquid chromatography technique. Asian J Pharm Clin Res 2018;11:110-3.

Shirkhedkar Atul A, Bhirud Charushila H, Surana Sanjay J. Application of UV-spectrophotometric methods for estimation of tenofovir disoproxil fumarate in tablets. Pak J Pharm Sci 2009;22:27-9.

Jullien V, Treluyer JM, Pons G, Rey E. Determination of tenofovir in human plasma by high-performance liquid chromatography with spectrofluorimetric detection. J Chromatogr B: Anal Technol Biomed Life Sci 2003;785:377-81.

Ashenafi D, Chintam V, Van Veghel D, Dragovic S, Hoogmartens J, Adams E. Development of a validated liquid chromatographic method for the determination of related substances and assay of tenofovir disoproxil fumarate. J Sep Sci 2010;33:1708-16.

Bodempudi SB, Rupakula RCB, Reddy KS. Development and validation of rp-chiral HPLC method for quantification of (s)-isomer in tenofovir disoproxil fumarate. Int J Curr Pharm Res 2017;9:31-6.

Agrahari V, Youan BB. Sensitive and rapid HPLC quantification of tenofovir from hyaluronic acid-based nanomedicine. AAPS PharmSciTech 2012;13:202-10.

Saha C, Gupta NV, Chandan RS, Shanmukha Priya P. The development of a validated stability indicating LC-MS method for the determination of tenofovir disoproxil fumarate using quality by design approach. Int J Appl Pharm 2019;11:406-17.

Wiriyakosol N, Puangpetch A, Manosuthi W, Tomongkon S, Sukasem C, Pinthong D. A LC/MS/MS method for determination of tenofovir in human plasma and its application to toxicity monitoring. J Chromatogr B: Anal Technol Biomed Life Sci 2018;1085:89-95.

Desai R, Roadcap B, Goykhman D, Woolf E. Determination of doravirine in human plasma using liquid-liquid extraction and HPLC-MS/MS. Bioanalysis 2019;11:1495-508.

International Conference on Harmonization (ICH). Stability testing of new drug substances and products Q1A (R2). Geneva, Switzerland; 2003.

International Conference on Harmonization (ICH). Harmonized tripartite guideline validation of analytical procedures: Text and methodology Q2 (R1). Geneva: IFPMA, Switzerland; 2005.

Ravichandran V, Shalini S, Sundaram KM, Rajak H. Validation of analytical methods-strategies and importance. Int J Pharm Pharm Sci 2010;2:18-22.



How to Cite

TIRUVEEDHI, V. L. N. B. G., BATTULA, V. R., & BONIGE, K. B. (2021). RP-HPLC (STABILITY-INDICATING) BASED ASSAY METHOD FOR THE SIMULTANEOUS ESTIMATION OF DORAVIRINE, TENOFOVIR DISOPROXIL FUMARATE AND LAMIVUDINE. International Journal of Applied Pharmaceutics, 13(1), 153–159. https://doi.org/10.22159/ijap.2021v13i1.39608



Original Article(s)