Rapid stability-indicating UHPLC method for determination of lamivudine and tenofovir disoproxil fumarate in fixed-dose combination tablets

Danielle Evangelista Rabelo de Souza; Nicoly Kaliny Magela Gomes; Gerson Antônio Pianetti; Bruno Gonçalves Pereira; Christian Fernandes

doi:10.22456/2527-2616.117905

Authors

Danielle Evangelista Rabelo de Souza Divisão de Controle de Qualidade, Diretoria Industrial - Fundação Ezequiel Dias
Nicoly Kaliny Magela Gomes Laboratório de Controle de Qualidade de Medicamentos e Cosméticos, Departamento de Produtos Farmacêuticos, Faculdade de Farmácia - Universidade Federal de Minas Gerais
Gerson Antônio Pianetti Laboratório de Controle de Qualidade de Medicamentos e Cosméticos, Departamento de Produtos Farmacêuticos, Faculdade de Farmácia - Universidade Federal de Minas Gerais
Bruno Gonçalves Pereira Divisão de Desenvolvimento de Medicamentos, Diretoria Industrial - Fundação Ezequiel Dias
Christian Fernandes Laboratório de Controle de Qualidade de Medicamentos e Cosméticos, Departamento de Produtos Farmacêuticos, Faculdade de Farmácia - Universidade Federal de Minas Gerais https://orcid.org/0000-0002-3905-3674

DOI:

https://doi.org/10.22456/2527-2616.117905

Keywords:

AIDS, antiviral drug, stability-indicating method, forced degradation, ultra high-performance liquid chromatography

Abstract

Lamivudine (3TC) and tenofovir disoproxil fumarate (TDF) are antiretroviral drugs widely used for AIDS treatment. Since safety, efficacy and quality are essential for drug products, stability studies must be performed. Therefore, degradation studies must be carried out to evaluate the degradation products formed. The active pharmaceutical ingredients 3TC and TDF, as well as the tablets containing the combination of these drugs were subjected to a comprehensive forced degradation. 3TC, TDF and the degradation products were analyzed by a stability-indicating ultra-high performance liquid chromatography (UHPLC) method developed and validated. A C8 (100 x 2.0 mm, 2.2 μm) column and a mobile phase composed of 0.1 M ammonium acetate buffer pH 4.0, acetonitrile and methanol in gradient elution, at 0.5 mL/min, were used. The injection volume was 4 μL and detection was at 260 nm. The method was selective, precise, accurate and linear in the range 0.1-0.6 mg mL-1 for the two drugs. 3TC was degraded in acidic, alkaline and oxidative environment and in the presence of metal ions. Two degradation products were observed. TDF was degraded in the same conditions than those 3TC was labile. In addition, TDF was degraded in neutral condition. Again, two degradation products were formed. Chemical structures were proposed for the degradation products using UHPLC-QTOF/MS. The stability-indicating method developed showed to be useful in stability studies and in the quality control of fixed-dose combination tablets containing 3TC and TDF.

Downloads

Download data is not yet available.

References

World Health Organization. HIV/AIDS. https://www.who.int/news-room/factsheets/detail/hiv-aids. Accessed 25 Aug 2021.

BRASIL. Ministério da Saúde. Secretaria de Vigilância em Saúde - Departamento de DST, AIDS e Hepatites Virais. Protocolo clínico e diretrizes terapêuticas para manejo da infecção pelo HIV em adultos, Brasília; 2015.

BRASIL. Ministério da Saúde. Secretaria de Vigilância em Saúde - Departamento de DST, AIDS e Hepatites Virais. Boletim Epidemiológico HIVAIDS, Brasília; 2015.

BRASIL. Ministério da Saúde. Secretaria de Vigilância em Saúde. Departamento de DST, AIDS e Hepatites virais. Adesão ao tratamento antirretroviral no Brasil: coletânea de estudos do projeto ATAR, Brasília; 2010.

Venkatesan S, Kannappan N, Mannemala SS. Stability-indicating HPLC method for the simultaneous determination of HIV tablet containing emtricitabine, tenofovir disoproxil fumarate, and rilpivirine hydrochloride in pharmaceutical dosage forms. Int Sch Res Notices 2014: 2014:1-9.

International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. Guideline Q1A(R2) - Stability Test of New Drug Substances and Products. Geneva; 2003.

BRASIL. Ministério da Saúde. Agência Nacional de Vigilância Sanitária. Resolução RDC nº 53, de 4 de dezembro de 2015, Brasília; 2015.

Swartz ME. UPLCTM: an introduction and review. J Liq Chromatogr Relat Technol 2005; 28:1253- 1263.

Bedse G, Kumar V, Singh S. Study of forced decomposition behavior of 3TC using LC, LCMS/TOF and MS. J Pharm Biomed Anal 2009: 49:55-63.

Gualdesi MS, Romero JE, Brinon MC, Raviolo MA. Development and validation of a stability indicating method for seven novel derivatives of 3TC with anti-HIV and anti-HBV activity in simulated gastric and intestinal fluids. J Pharm Biomed Anal 2013; 78-79:52-56.

Navaneethan G, Karunakaran K, Elango KP. Specific and selective quantification of antiviral drugs in the presence of impurities by liquid chromatography. J Food Drug Anal 2012; 20:446- 453.

Kaul N, Agrawal, H, Paradkar AR, Mahadik, KR. The International Conference on Harmonisation guidance in practice: Stress degradation studies on lamivudine and development of a validated specific stability-indicating HPTLC assay method. Chromatographia 2004; 60:213-221.

Ashenafi D, Chintam V, Veghel DV, 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; 33:1708-1716.

Golla VM, Kurmi M, Shaik K, Singh S. Stability behaviour of antiretroviral drugs and their combinations. 4: Characterization of degradation products of tenofovir alafenamide fumarate and comparison of its degradation and stability behavior with tenofovir disoproxil fumarate. J Pharm Biomed Anal 2016; 131:146–155.

Kurmi M, Kushwah BS, Sahu A, Narayanam M, Singh S. Stability behavior of antiretroviral drugs and their combinations. 2: Characterization of interaction products of 3TC and tenofovir disoproxil fumarate by mass and NMR spectrometry. J Pharm Biomed Anal 2016; 125:245-259.

Krishna MVVNM, Rao, SV, Venugopal NVS. Forced degradation study for tenofovir df, lamivudine and efavirenz in triple combination antiretroviral tablets and development of validated stability indicating assay method by UPLC. Curr Pharm Anal 2019; 15:82-94.

BRASIL. Agência Nacional de Vigilância Sanitária Resolução nº 899, de 29 de maio de 2003 - Guia para validação de métodos analíticos e bioanalíticos, Brasília; 2003.

Souza, SVC. Procedimento para validação intralaboratorial de métodos de ensaio: delineamento e aplicabilidade em análise de alimentos [PhD thesis], Belo Horizonte (MG): Universidade Federal de Minas Gerais; 2007.

Souza SVC, Junqueira RG. A procedure to assess linearity by ordinary least squares method. Anal Chim Acta 2005; 552:25-35.

Carstensen JT, Rhodes CT. Drug stability. Principles and Practices. New York: Marcel Dekker; 2000.

Snyder LR, Kirkland JJ, Glajch JL. Practical HPLC method development. New York: John Wiley & Sons; 1997.

Green JM. A practical guide to analytical method validation. Anal Chem 1996; 68:305A-309A.

Youden WJ, Steiner EH. Statistical manual of the Association of Official Analytical Chemists. 48th ed. Arlington: AOAC, 1975.

World Health Organization. The International Pharmacopoeia, seventh ed; 2017.

Silva JPA. Estudo de estabilidade do antirretroviral tenofovir: uma abordagem integrada para o desenvolvimento de medicamento [dissertation]. Recife (PE): Universidade Federal de Pernambuco; 2014.