Forced Degradation Study for Tenofovir DF, Lamivudine and Efavirenz in Triple Combination Anti-Retroviral Tablets and Development of Validated Stability Indicating Assay Method by UPLC

Author(s): Mantripragada V.V.N. Murali Krishna*, Sumathi Vinay Rao, Nutulapati V.S. Venugopal

Journal Name: Current Pharmaceutical Analysis

Volume 15 , Issue 1 , 2019


Become EABM
Become Reviewer
Call for Editor

Graphical Abstract:


Abstract:

Introduction: Tenofovir Disoproxil Fumarate/Lamivudine/Efavirenz Tablets are a pharmaceutical dosage form indicated for the treatment of Human Immunodeficiency Virus (HIV) infection.

Methods: A simple, time efficient and stability indicating Reverse Phase Ultra Performance Liquid Chromatography (RP – UPLC) method was developed for the simultaneous determination of the three drugs present in the tablets. Mobile phase-A is 0.1% Trifluoro Acetic acid in water and mobile phase-B is acetonitrile in gradient elution mode. Flow rate at 0.4 mL/min was fixed in the method. Acquity BEH Phenyl (100 mm × 2.1 mm), 1.7 µ column was used for the separation of analytes at 260 nm wavelength. The developed method has been validated as specified in International Conference on Harmonization (ICH) guideline. Forced degradation study was conducted for Tenofovir Disoproxil fumarate (TDF), Lamivudine (LAM) and Efavirenz (EFA) pharmaceutical tablets to identify the degradation behaviour of individual drugs under stress study.

Results: The method was observed to be linear from 6-90 µg/mL concentration for TDF & LAM and 12-180 µg/mL for EFA with correlation coefficient values of more than 0.999. The method was found to be precise with RSD (Relative standard deviation) values of below 2% for replicate measurements. Recovery results were found to be between 99.0 and 100.8%. LOQ values for TDF, LAM & EFA were 0.041 µg/mL, 0.034 µg/mL & 0.053 µg/mL, respectively. Specificity of the analytical method was checked by injecting the stressed samples and evaluating the homogeneity of peaks using Photo Diode Array (PDA) detector. Peak purity results indicated that no interference was observed from degradation impurities for the quantification of TDF, LAM and EFA in the dosage form. The method was found to be robust under the deliberately modified chromatographic parameters.

Conclusion: The developed UPLC method is stability indicating, successfully validated and suitable for quantification purpose.

Keywords: Tenofovir disoproxil fumarate, lamivudine, efavirenz, forced degradation, stability indicating and UPLC, antiretroviral tablets.

[1]
Drug bank: The DrugBank database is a comprehensive, freely accessible, online database containing information on drugs and drug targets.. https://www.drugbank.ca/drugs/DB00300
[2]
Moore, R.D. Natural history of HIV infection in the era of combination antiretroviral therapy. Chaisson RE. AIDS, 1999, 13(14), 1933-1942.
[3]
Raza, M.S.A.; AlOthman, Z.A.; Nafisur, R. Analytical techniques in pharmaceutical analysis: A review. Arabian . J. Chem., 2017, 10(1), S1409-S1421.
[4]
AlOthman, Z.A.; Rahman, N.; Siddiqui, M.R. Review on pharmaceutical impurities, stability studies and degradation products. Rev. Adv. Sci. Eng., 2013, 2(2), 155-166.
[5]
Rahman, N.; Azmi, S.N.H.; Wu, H.F. The importance of impurity analysis in pharmaceutical products: an integrated approach. Accred. Qual. Assur., 2006, 11(1-2), 69-74.
[6]
Bhavsar, S.D.; Patel, N.B.; Patel, N.C. RP-HPLC method for simultaneous estimation of Tenofovir disoproxil fumarate, Lamivudine, and Efavirenz in combined tablet dosage form. Pharm. Methods, 2012, 3(2), 73-78.
[7]
Kumar, K.A.; Abirami, G.; Murugan, S.; Ashok, B. RP-HPLC method for simultaneous estimation of Lamivudine, Tenofovir disoproxil fumarate and Efavirenz in tablet formulation. J. Anal. Chem., 2013, 68(9), 815-821.
[8]
Srinath, A.; Sneha, B.; Akhila, A.; Ahmed, R.; Kulkarni, R.G. Method development and validation for simultaneous estimation of lamivudine, tenofovir and efavirenz in combined tablet dosage form by RP-HPLC and UV-Spectroscopic Method. Int. J. Pharm. Sci. Res., 2014, 5(12), 5491-5497.
[9]
Bhargavi, G. Reddy.; K Mohammad, Younus. T.; Ravindra Reddy, Y. Method development and validation of Lamivudine, Tenofovir and Efavirenz in a combined dosage form by RP-HPLC. J. Pharm. Res., 2012, 5(12), 711-714.
[10]
Vanaja, P.; Anusha, N.; Giri Prasad, V.S. Development and validation of a RP-HPLC method for simultaneous estimation of Lamivudine, Tenofovir Disoproxil Fumarate and Efavirenz in a combined tablet dosage form. Int. J. Pharm. Pharm. Sci., 2013, 5(3), 116-121.
[11]
Rao, T.M.; Prabhakar, T.; Girija Sankar, G.; Naidu, P.V.L.; Jyothi, N. Simultaneous determination of Lamivudine, Tenofovir and Efavirenz in Lamivudine, Tenofovir Dispropoxil Fumarate and Efavirenz tablets by stability indicating isocratic RP-HPLC method with PDA detector. Int. J. Pharm., 2014, 3(10), 270-276.
[12]
Rao, N.S.; Venkataramana, K.; Srinivas, P.; Meghana, R.; Anitha, P. Method development and validation for simultaneous estimation of Lamivudine, Tenofovir Disoproxil Fumarate and Efavirenz in combined tablet dosage form by RP-HPLC. World J. Pharm. Pharmaceut. Sci., 2013, 3(1), 1658-1671.
[13]
Sharma, R.; Mehta, K. Simultaneous spectrophotometric estimation of tenofovir disoproxil fumarate and lamivudine in three component tablet formulation containing efavirenz. Indian J. Pharm. Sci., 2010, 72(4), 527-530.
[14]
Vidyadhara, S.; Sasidhar, R.L.C.; Venkateswara Rao, B.; Ratna Kumari, P. Simultaneous UV spectrophotometric method for the determination of tenofovir, efavirenz and lamivudine in bulk and combined dosage form. Asian J. Pharmaceut. Anal, 2016, 6(4), 253-258.
[15]
Murugan, S.; Sikdar, P.; Debnath, S.; Niranjan Babu, M. Development and validation of first order Derivative UV spectrophotometric method for the Estimation of Tenofovir disoproxil fumarate, Lamivudine and Efavirenz in bulk and tablet dosage Form. Int. J. Pharmaceut. Res. Anal., 2013, 3(1), 29-32.
[16]
Reddiah, C.H.V.; Devi, P.R.; Mukkanti, K.; Srinivasu, P. Development and validation of stability indicating hplc method for combination tablet dosage form of efavirenz, lamivudine and tenofovir in tablet. Int. J. Pharm. Phytopharm. Res, 2012, 2(1), 40-45.
[17]
SkIsmail. Y.M.; Gunasekaran, V. Development and validation for simultaneous estimation of Lamivudine, Tenofovir and Efavirenz by UPLC. Int. J. Pharm., 2012, 2(3), 656-660.
[18]
Rahman, N.; Sameen, S.; Kashif, M. Spectroscopic study on the interaction of haloperidol and 2,4-dinitrophenylhydrazine and its application for the quantification in drug formulations. Analyt. Chem. Lett., 2016, 6(6), 874-885.


Rights & PermissionsPrintExport Cite as

Article Details

VOLUME: 15
ISSUE: 1
Year: 2019
Published on: 28 November, 2018
Page: [82 - 94]
Pages: 13
DOI: 10.2174/1573412913666171120163959
Price: $65

Article Metrics

PDF: 26
HTML: 4
EPUB: 1