Back
 AJAC  Vol.6 No.10 , September 2015
Stability Indicating RP-UPLC Method for Assay of Emtricitabine and Tenofovir Disoproxil Fumarate in Bulk and Dosage Forms
Abstract: A simple, sensitive and rapid stability indicating reverse phase ultra performance liquid chromatography (RP-UPLC) method was developed and validated for the determination of Emtricitabine (EMT) and Tenofovir disoproxil fumarate (TDF) in pure and tablet dosage forms. The chromatographic separation was achieved by using Waters (Alliance) UPLC system equipped with auto-sampler and photo diode array detector. A volume of 5 μL of standard or test was injected into the column and the components were separated by using the mixture of 0.68% potassium dihydrogen orthophosphate buffer of pH = 6 and methanol in the ratio 45:55 v/v as mobile phase at a flow rate of 1.2 mL/min through BEH C18 (100 mm × 2.1, 1.8 μm) at ambient temperature and were detected at a wavelength of 261 nm. System suitable parameters such as plate count and tailing factor for EMT and TDF were found to be 2427 & 3685, 1.16 & 1.23 respectively, and resolution between EMT and TDF peaks was found to be 3.12. The chromatographic parameters like retention time, peak area and peak height of EMT and TDF were found to be 0.684 & 0.930, 694,200 & 8,778,000 and 272,881 & 3685 respectively. Percent of assay of EMT and TDF in bulk and dosage forms was determined and found to be 101.48 and 103.22 respectively. Study of degradation was examined and found that the drugs were stable under degradation conditions. The present method was developed keeping the principles of green chemistry by using eco-friendly solvent methanol in mobile phase. The developed method was found to be simple, rapid and applied for the analysis of Truvada; therefore the proposed method is recommended for the analysis of EMT and TDF in pure and tablet dosage forms in any quality control laboratories.
Cite this paper: Purnima, B. , Reddy, T. , Rao, Y. , Ramu, G. and Ramachandran, D. (2015) Stability Indicating RP-UPLC Method for Assay of Emtricitabine and Tenofovir Disoproxil Fumarate in Bulk and Dosage Forms. American Journal of Analytical Chemistry, 6, 807-821. doi: 10.4236/ajac.2015.610077.
References

[1]   Anandakumar, K., Kannan, K. and Vetrichelvan, T. (2011) Development and Validation of Emtricitabine and Tenofovir Disoproxil Fumerate in Pure and in Fixed Dose Combination by UV Spectrophotometry. Digest Journal of Nano-materials and Biostructures, 6, 1085-1090.

[2]   Patel, S., Baghel, U.S., Rajesh, P., Prabhakar, D., Engla, G. and Nagar, P.N. (2009) Spectrophotometric Method Development Tenofovir Disoproxil Fumarate and Emtricitabine in Bulk Drug and Tablet Dosage Form. International Journal of Pharmaceutical and Clinical Research, 1, 28-30.

[3]   Choudhari, V.P., Ingale, K.D., Barhate, A., Kale, A.N., Bobade, C.D. and Kuchekar, B.S. (2010) Development and Validation of Simultaneous and Isoabsorptive UV-Spectrophotometric Methods for Tenofovir and Emtricitabine in Pharmaceutical Formulations. Journal of Pharmacy Research, 9, 11-13.

[4]   Ghorpade, S.A., Sali, M.S., Kategaonkar, A.H., Patel, D.M., Choudhari, V.P. and Kuchekar, B.S. (2010) Simultaneous Determination of Emtricitabine and Tenofovir by Area under Curve and Dual Wavelength Spectrophotometric Method. Journal of the Chilean Chemical Society, 54, 331-333.
http://dx.doi.org/10.4067/s0717-97072010000100027

[5]   Vishnu, P.C., Snehal, I., Sacchidanand, R.G., Dipali, D.T., Vikram, G.M. and Ambekar, A. (2011) Spectrophotometric Simultaneous Determination of Tenofovir Disoproxil Fumarate and Emtricitabine in Combined Tablet Dosage Form by Ratio Derivative, First Order Derivative and Absorbance Corrected Methods and Its Application to Dissolution Study. Pharmaceutical Methods, 2, 47-52.
http://dx.doi.org/10.4103/2229-4708.81096

[6]   Ahindita, P., Aurobinda, K.M., Amit, G.S., Dannana, K., Swapna, M. and Sudam, C.S.I. (2010) Development and Validation of Spectrophotometric Methods for Determination of Emtricitabine and Tenofovir Disoproxil Fumarate in Bulk and Tablet Dosage Form. International Journal of PharmTech Research, 3, 1874-1882.

[7]   Ghorpade, S.A., Sali, K.A.H., Patel, D.M., Choudhari, V.P. and Kuchekar, B.S. (2010) Simultaneous Determination of Emtricitabine and Tenofovir by Area under Curve and Dual Wavelength Spectrophotometric Method. Journal of the Chilean Chemical Society, 55, 115-117.
http://dx.doi.org/10.4067/S0717-97072010000100027

[8]   Choudhari, V.P., Ingale, S., Gite, S.R., Tajane, D.D., Modak, V.G. and Ambekar, A. (2011) Spectrophotometric Simultaneous Determination of Tenofovir Disoproxil Fumarate and Emtricitabine in Combined Tablet Dosage Form by Ratio Derivative First Order Derivative and Absorbance Corrected Methods and Its Application to Dissolution Study. Pharmaceutical Methods, 1, 47-52.
http://dx.doi.org/10.4103/2229-4708.81096

[9]   Karunakaran, A., Kamarajan, K. and Thangarasu, V. (2010) A Validated RP-HPLC Method for Simultaneous Estimation of Emtricitabine and Tenofovir Disoproxil Fumarate in Pure and in Tablet Dosage Form. Pharm Sin, 1, 52-60.

[10]   Sharma, R. and Gupta, P. (2009) A Validated RP-HPLC Method for Simultaneous Estimation of Emtricitabine and Tenofovir Disoproxil Fumarate in a Tablet Dosage Form. Eurasian Journal of Analytical Chemistry, 4, 276-284.

[11]   Pranitha, D., Vanitha, C. and Francies, P. (2010) Simultaneous Estimation of Emtricitabine, Tenofovir Disoproxil Fumarate and Rilpivirine in Bulk Form by RP-HPLC Method. Journal of Pharmacy Research, 5, 4600-4602.

[12]   Raju, N.A. and Begum, S. (2008) Simultaneous RP-HPLC Method for the Estimation of the Emtricitabine, Tenofovir Disoproxil Fumarate and Efavirenz in Tablet Dosage Forms. Research J Pharm Technol, 1, 522-525.

[13]   Rezk, N.L., Crutchley, R.D. and Kashuba, A.D. (2005) Simultaneous Quantification of Emtricitabine and Tenofovir in Human Plasma Using High-Performance Liquid Chromatography after Solid Phase Extraction. Journal of Chromatography B, 822, 201-208.
http://dx.doi.org/10.1016/j.jchromb.2005.06.019

[14]   Nirogi, R., Bhyrapuneni, G., Kandikere, V., Mudigonda, K., Komarneni, P., Aleti, R., et al. (2009) Simultaneous Quantification of a Non-Nucleoside Reverse Transcriptase Inhibitor Efavirenz, a Nucleoside Reverse Transcriptase Inhibitor Emtricitabine and a Nucleotide Reverse Transcriptase Inhibitor Tenofovir in Plasma by Liquid Chromatography Positive Ion Electrospray Tandem Mass Spectrometry. Journal of Chromatography, 23, 371-381.
http://dx.doi.org/10.1002/bmc.1125

[15]   Gomes, N.A., Vaidya, V.V., Pudage, A., Joshi, S.S. and Parekh, S.A. (2008) Liquid Chromatography-Tandem Mass Spectrometry Method for Simultaneous Determination of Tenofovir and Emtricitabine in Human Plasma and Its Application to a Bioequivalence Study. Journal of Pharmaceutical and Biomedical Analysis, 48, 918-926.
http://dx.doi.org/10.1016/j.jpba.2008.07.022

[16]   Delahunty, T., Bushman, L., Robbins, B. and Fletcher, C.V. (2009) The Simultaneous Assay of Tenofovir and Emtricitabine in Plasma Using LC/MS/MS and Isotopically Labeled Internal Standards. Journal of Chromatography B, 877, 1907-1914.
http://dx.doi.org/10.1016/j.jchromb.2009.05.029

[17]   Avolio, A.D., Sciandra, M. and Siccardi, M. (2008) A New Assay Based on Solid-Phase Extraction Procedure with LC-MS to Measure Plasmatic Concentrations of Tenofovir and Emtricitabine in HIV Infected Patients. Journal of Chromatographic Science, 46, 524-528.
http://dx.doi.org/10.1093/chromsci/46.6.524

[18]   Yadav, M., Singhal, P., Goswami, S., Pande, U.C., Sanyal, M. and Shrivastav, P.S. (2010) Selective Determination of Antiretroviral Agents Tenofovir, Emtricitabine, and Lamivudine in Human Plasma by a LC-MS-MS Method for a Bioequivalence Study in Healthy Indian Subjects. Journal of Chromatographic Science, 48, 704-713.
http://dx.doi.org/10.1093/chromsci/48.9.704

[19]   Joshi, M., Nikalje, A.P., Shahed, M. and Dehghan, M. (2009) HPTLC Method for the Simultaneous Estimation of Emtricitabine and Tenofovir in Tablet Dosage Form. Indian Journal of Pharmaceutical Sciences, 71, 95-97.
http://dx.doi.org/10.4103/0250-474X.51951

[20]   Kavitha, K.Y., Geetha, G., Hariprasad, R., Venkatnarayanan, R. and Kaviarasu, M. (2013) Development and Validation of RP-UPLC Analytical Method for Simultaneous Estimation of the Emtricitabine, Tenofovir Disoproxil Fumarate and Rilpivirine and Its Pharmaceutical Dosage Form. International Research Journal of Pharmacy, 4, 150-155.

[21]   Nagaraju, P.T., Channabasavaraj, K.P. and Shantha Kumar, P.T. (2011) Development and Validation of Spectrophotometric Method for Estimation of Emtricitabine in Tablet Dosage Form. International Journal of ChemTech Research, 3, 23-28.

[22]   Droste, J.A.H., Aarnoutse, R.E. and Burger, D.M. (2007) Determination of Emtricitabine in Human Plasma Using HPLC with Fluorometric Detection. Journal of Liquid Chromatography & Related Technologies, 30, 2769-2778.
http://dx.doi.org/10.1080/10826070701560900

[23]   Ashenafi, D., Verbeek, A., Hoogmartens, J. and Adams, E. (2009) Development and Validation of an LC Method for the Determination of Emtricitabine and Related Compounds in the Drug Substance. Journal of Separation Science, 32, 1823-1830.
http://dx.doi.org/10.1002/jssc.200800688

[24]   Seshachalam, U., Haribabu, B. and Chandrasekhar, K.B. (2007) Development and Validation of a Stability-Indicating Liquid Chromatographic Method for Determination of Emtricitabine and Related Impurities in Drug Substance. Journal of Separation Science, 30, 999-1004.
http://dx.doi.org/10.1002/jssc.200600429

[25]   Pendela, M., Mamade, D.A., Hoogmartens, J., Van Schepdael, A. and Adams, E. (2010) Characterization of Emtricitabine Related Substances by Liquid Chromatography Coupled to an Ion Trap Mass Spectrometer. Talanta, 82, 125-128.
http://dx.doi.org/10.1016/j.talanta.2010.04.004

[26]   Atul, H.B. and Charashila Sajai, J.S. (2009) Application of UV-Spectrophotometric Methods for Estimation of Tenofovir Disoproxil Fumarate in Tablets. Pakistan Journal of Pharmaceutical Sciences, 22, 27-29.

[27]   Pratap Reddy, J. and Chakravarthy, I.E. (2012) New Spectrophotometric Determination of Tenofovir Disoproxil Fumaratein Bulk and Pharmaceutical Dosage Form. IOSR Journal of Applied Chemistry, 1, 29-33.
http://dx.doi.org/10.9790/5736-0122933

[28]   Rani, T.S., Sujatha, K. and Chitra, K. (2012) Spectrophotometric Methods for Estimation of Tenofovir Disoproxil Fumarate in Tablet. J Pharm Anal, 1, 9-12.

[29]   Onah, J.O. and Ajima, U. (2011) Spectrophotometric Determination of Tenofovir Disoproxil Fumarate after Complexation with Ammonium Molybdate and Picric Acid. International Journal of Drug Development and Research, 3, 199-204.

[30]   Kandagal, P.B., Manjunatha, D.H., Seetharamappa, J. and Kalanur, S.S. (2008) RP-HPLC Method for the Determination of Tenofovir in Pharmaceutical Formulations and Spiked Human Plasma. Analytical Letters, 41, 561-570.
http://dx.doi.org/10.1080/00032710801910742

[31]   Jullien, V., Tréluyer, J.M., Pons, G. and Rey, E. (2003) Determination of Tenofovir in Human Plasma by High-Performance Liquid Chromatography with Spectrofluorimetric Detection. Journal of Chromatography B, 785, 377-381.
http://dx.doi.org/10.1016/S1570-0232(02)00933-9

[32]   Sparidans, R.W., Crommentuyn, K.M.L., Schellens, J.H.M. and Beijnen, J.H. (2003) Liquid Chromatographic Assay for the Antiviral Nucleotide Analogue Tenofovir in Plasma Using Derivatization with Chloroacetaldehyde. Journal of Chromatography B, 791, 227-233.
http://dx.doi.org/10.1016/S1570-0232(03)00225-3

[33]   Bhavsar, S., Patel, B.N. and Patel, C.N. (2012) RP-HPLC Method for Simultaneous Estimation of Tenofovir Disoproxil Fumarate, Lamivudine, and Efavirenz in Combined Tablet Dosage Form. Pharm Methods, 3, 73-78.
http://dx.doi.org/10.4103/2229-4708.103876

[34]   Barkil, M.E., Gagnieu, M. and Guitton, J. (2007) Relevance of a Combined UV and Single Mass Spectrometry Detection for the Determination of Tenofovir in Human Plasma by HPLC in Therapeutic Drug Monitoring. Journal of Chromatography B, 54, 192-197.
http://dx.doi.org/10.1016/j.jchromb.2007.04.015

[35]   Sentenac, S., Fernandez, C., Thuillier, A., Lechat, P. and Aymard, G. (2003) Sensitive Determination of Tenofovir in Human Plasma Samples Using Reversed-Phase Liquid Chromatography. Journal of Chromatography B, 793, 317-324.
http://dx.doi.org/10.1016/S1570-0232(03)00333-7

 
 
Top