Author(s):
Sathish Kumar Konidala, Ramu Samineni, Yamarthi Venkateswara Rao, Jithendra Chimakurthy, Chaitanya Sucharitha Kolakaluri, Vishali Korrapati
Email(s):
sathishkonidala@gmail.com
DOI:
10.52711/0974-360X.2022.00525 
Address:
Sathish Kumar Konidala*, Ramu Samineni, Yamarthi Venkateswara Rao, Jithendra Chimakurthy, Chaitanya Sucharitha Kolakaluri, Vishali Korrapati
Faculty of Pharmacy, Department of Pharmaceutical Sciences, Vignan’s Foundation for Science, Technology and Research, Vadlamudi, Guntur, Andhra Pradesh-522213.
*Corresponding Author
Published In:
Volume - 15,
Issue - 7,
Year - 2022
ABSTRACT:
Objective: The objective of this work is to develop a precise, accurate and validated reverse phase ultra-performance liquid chromatographic technique for effective simultaneous determination of Emtricitabine, Tenofovir and Efavirenz in bulk and tablet formulation. Method: Separation of the selected drugs was optimized after several trials including changing mobile phase and its composition, stationary phase, flow rate, column temperature, etc. Finally the separation of drugs was achieved on BEH C18 column using a mixture of methanol and phosphate buffer having pH 3.5 in the ratio of 65:35 v/v as mobile phase with flow rate of 0.3 ml/min and the analytes were detected at a wavelength of 260 nm. Results: The developed method was validated by determining the parameters like linearity, system suitability, recovery, precision, specificity, robustness, ruggedness, LOD, and LOQ as per the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines. The system suitability parameters were within the limits, retention times (Rt) for Emtricitabine, Tenofovir and Efavirenz were found to be 0.432, 0.671, and 2.772 min respectively. The method showed linearity between the concentration range of 10-50 µg/ml for Emtricitabine (r2= 0.9987), 15-75 µg/ml for Tenofovir (r2= 0.9983) and 30-150 µg/ml for Efavirenz (r2= 0.9982). The percentage recovery results at 50%, 100% and 150% of Emtricitabine, Tenofovir and Efavirenz were found to be in the range of 99.45 % - 100.15 %. Since there was no interference due to excipients and mobile phase, the method was found to be specific. The assay results of the combined tablet dosage form by the developed method were identified as in good agreement with the acceptance limit. Conclusions: The current method was proved to be effective for routine simultaneous determination of Emtricitabine, Tenofovir and Efavirenz in bulk and tablet formulation.
Cite this article:
Sathish Kumar Konidala, Ramu Samineni, Yamarthi Venkateswara Rao, Jithendra Chimakurthy, Chaitanya Sucharitha Kolakaluri, Vishali Korrapati. Novel RP-UPLC Method Development and Validation for Simultaneous Quantification of Emtricitabine, Tenofovir and Efavirenz in Bulk and Tablet Dosage Forms. Research Journal of Pharmacy and Technology. 2022; 15(7):3141-6. doi: 10.52711/0974-360X.2022.00525
Cite(Electronic):
Sathish Kumar Konidala, Ramu Samineni, Yamarthi Venkateswara Rao, Jithendra Chimakurthy, Chaitanya Sucharitha Kolakaluri, Vishali Korrapati. Novel RP-UPLC Method Development and Validation for Simultaneous Quantification of Emtricitabine, Tenofovir and Efavirenz in Bulk and Tablet Dosage Forms. Research Journal of Pharmacy and Technology. 2022; 15(7):3141-6. doi: 10.52711/0974-360X.2022.00525 Available on: https://www.rjptonline.org/AbstractView.aspx?PID=2022-15-7-47
REFERENCES:
1. https://www.rxlist.com/emtriva-drug.htm#description(Accessed on 28thjune 2020)
2. https://pubchem.ncbi.nlm.nih.gov/compound/Tenofovir(Accessed on 28thjune 2020)
3. https://www.rxlist.com/viread-drug.htm(Accessed on 30thjune 2020)
4. Cristofoletti R, Nair A, Abrahamsson B, Groot DW, Kopp S, Langguth P, Polli JE, Shah VP, Dressman JB. Biowaiver monographs for immediate release solid oral dosage forms,efavirenz, Journal of pharmaceutical sciences. (2013)102(2)318-29.
5. Jacob J, Nadig S. A UPLC method for simultaneous estimation of emtricitabine, tenofovir disoproxil fumarate and efavirenz in pharmaceutical dosage forms. Research Journal of Pharmacy and Technology. (2017) 10(12)4463-6.
6. El Barkil M, Gagnieu MC, Guitton J. 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, (2007)854(1-2)192-7.
7. Anbazhagan S, Indumathy N, Shanmugapandiyan P, Sridhar SK. Simultaneous quantification of stavudine, lamivudine and nevirapine by UV spectroscopy, reverse phase HPLC and HPTLC in tablets. Journal of pharmaceutical and biomedical analysis. (2005) 39(3-4)801-4.
8. Shewiyo DH, Kaale E, Ugullum C, Sigonda MN, Risha PG, Dejaegher B, Smeyers–Verbeke J, Vander Heyden Y. Development and validation of a normal-phase HPTLC method for the simultaneous analysis of lamivudine, stavudine and nevirapine in fixed-dose combination tablets. Journal of pharmaceutical and biomedical analysis, (2011) 54(3)445-50.
9. Shelke A, Shinde M, Mogal R, Sable R, Jadhav A. Application of UV Spectrophotometric methods for simultaneous estimation of Emtricitabine and Tenofoviralafenamide fumarate in bulk. Asian Journal of Pharmacy and Technology. 2018;8(2):103-7.
10. Rao DS, Balaji M, Reddy SH, Ramu S, Rao KK, Krishna GR. Development and UV-spectrophotometric Methods for Quantitative Estimation of Tenofovirdisproxilfumarate in Bulk and Pharmaceutical Dosage Form. Asian Journal of Research in Chemistry. 2012;5(3):422-4.
11. Vidyadhara S, Sasidhar RL, Rao BV, Kumari PR. Simultaneous UV Spectrophotometric Method for the Determination of Tenofovir, Efavirenz and Lamivudine in Bulk and Combined Dosage Form. Asian Journal of Pharmaceutical Analysis. 2016;6(4):253-8.
12. Montgomery ER, Edmanson AL, Cook SC, Hovsepian PK. Development and validation of a reverse-phase HPLC method for analysis of efavirenz and its related substances in the drug substance and in a capsule formulation, Journal of pharmaceutical and biomedical analysis. 200125(2)267-84.
13. Kappelhoff BS, Rosing H, Huitema AD, Beijnen JH. Simple and rapid method for the simultaneous determination of the non-nucleoside reverse transcriptase inhibitors efavirenz and nevirapine in human plasma using liquid chromatography, Journal of Chromatography B. 2003 Jul 25;792(2):353-62.
14. Jayaseelan S, Ganesh S, Rajasekar M, Sekar V, Perumal P. A new analytical method development and validation for the simultaneous estimation of lamivudine and stavudine in tablet dosage form by RP-HPLC method. Int J Pharm Tech Res. ((2010) 2(2)1539-42.
15. Jayakar B, Kumar M, Saravanan C, Kumudhavalli MV. Method development and validation of RP-HPLC method for simultaneous determination of Lamivudine and Zidovudine. J Chem Pharm Res. (2010)2(1)478-81.
16. Sharma R, Gupta P. Simultaneous quantification and validation of emtricitabine and tenofovirdisoproxil fumarate in a tablet dosage form. Eurasian journal of analytical chemistry. (2009) 4(3)276-84.
17. Jayaseelan S, Suresh S, Sathishkumar G, Sekar V, Perumal P. Bioanalytical method development and validation of Lamivudine by RP-HPLC method. Int J Chem Tech Res. (2010) 2163-7.
18. Rao BU, Nikalje AP. Stability-indicating HPLC method for the determination of efavirenz in bulk drug and in pharmaceutical dosage form. African journal of Pharmacy and Pharmacology. (2009) Dec 313(12)643-50.
19. Malipatil SM, Nandedkar MA. Determination of TenofovirDisoproxilFumarate by a Sensitive Simple Isocratic RP-HPLC Method, Journal of Indian Council of Chemists. (2009) 26(1):67-9.
20. Mandloi DK, Tyagi PK, Rai VK, Dey S, Ashada RK, Mohanraj P. Method development and validation of RP-HPLC in the application of in vitro dissolution study of lamivudine in bulk drug and tablet formulation, J Chem Pharm Res. (2009) 1286-96.
21. Sarat M, Krishna PM, Rambabu C. Development and Validation of RP-UPLC Method for Simultaneous Estimation of AbacavirSulphate and Lamivudine in Combined Tablet Dosage Form, International Journal of ChemTech Research. 2012 Jul3(4)939-44.
22. Patel BN, Suhagia BN, Patel CN. Development and Validation of HPLC Method for the Estimation of Emtricitabine in Capsule Dosage Form. Asian Journal of Research in Chemistry. 2010;3(4):869-71.
23. Rao BS, Nagaraju S, Kiran BV. Development, Validation and Stress Degradation Studies of Emtricitabine and TenofovirDisoproxilFumerate by High Performance Liquid Chromatography. Asian Journal of Research in Chemistry. 2013;6(10):936-44.
24. Sreelatha P, Devi BR. Development and Validation of Stability-indicating HPLC method for simultaneous estimation of Tenofovir, Emtricitabine and Efavirenz in fixed dose combination drug product. Asian Journal of Research in Chemistry. 2018;11(1):23-31.
25. Shaheen R, Rizwan SH. A Stability Indicating Method for estimating Emtricitabine and Tenofovirdisoproxil fumarate simultaneously in Bulk and Combined dosage form by RP-HPLC. Asian Journal of Research in Chemistry. 2020;13(3):203-8.
26. Sundar BS, Balamuralikrishna K. Development and Validation of New Reversed Phase High Performance Liquid Chromatography Method for the Estimation of Efavirenz in Bulk and in Pharmaceutical Dosage Forms. Asian Journal of Research in Chemistry. 2011;4(4):555-7.
27. Raju NA, Begum S. Simultaneous RP-HPLC method for the estimation of the emtricitabine, tenofovirdisoproxilfumerate and efavirenz in tablet dosage forms. Research Journal of Pharmacy and Technology. 2008;1(4):522-5.
28. Jacob J, Nadig S. A UPLC method for simultaneous estimation of emtricitabine, tenofovirdisoproxilfumarate and efavirenz in pharmaceutical dosage forms. Research Journal of Pharmacy and Technology. 2017;10(12):4463-6.
29. Gomes NA, Vaidya VV, Pudage A, Joshi SS, Parekh SA. Liquid chromatography–tandem mass spectrometry (LC–MS/MS) method for simultaneous determination of tenofovir and emtricitabine in human plasma and its application to a bioequivalence study, Journal of Pharmaceutical and Biomedical Analysis. (2008) Nov 448(3)918-26.
30. Prathipati PK, Mandal S, Destache CJ. LC–MS/MS method for the simultaneous determination of tenofovir, emtricitabine, elvitegravir and rilpivirine in dried blood spots. Biomedical Chromatography. (2018) Sep32(9) e4270.
31. US Food and Drug Administration, “ICH Guidelines on Validation of Analytical Procedures, Text and Methodology Q2 (R1),” Federal Register, FDA, Silver Spring, (2005).
32. Jacob J, Nadig S. A UPLC method for simultaneous estimation of emtricitabine, tenofovirdisoproxilfumarate and efavirenz in pharmaceutical dosage forms. Research Journal of Pharmacy and Technology. (2017)10(12)4463-6.