METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF ENTECAVIR IN BULK AND PHARMACEUTICAL DOSAGE FORMS BY RP-HPLC
Objective: The objective and purpose of the analysis have sensibly assessed by selecting of a rapid and sensitive RP-HPLC method for Entecavir in bulk and pharmaceutical dosage form by using the most commonly employed C-18column with UV detection.
Methods: In estimation by RP-HPLC method Agilent 1120 compact LC system with variable programmable UV detector and Rheodyne injector with 20 Âµl fixed loop was used for the chromatographic separation. The mode of operation was isocratic with the components of a solution consisting of methanol: acetonitrile(70:30v/v) and triethanolamine (2-4drops)at the flow rate of 1.2 ml/min and run time was 10 min. Forced degradation studies were conducted to evaluate the stability and specificity of the method along with the validation parameters.
Results: Validation parameters of HPLC were found at a detection wavelength of 255 nm. Linearity was observed with the concentration range (Beerâ€™s law range) 20-100Âµg/ml with R2=0.9991. Robustness with detection wavelengths 253 and 257 nm with a flow rate of 1 ml/min and 1.4 ml/min showed good results. The retention time of the drug was 2.64 min and assay showed 98.1%.
Conclusion: The proposed RP-HPLC method was validated as per the ICH Q2B Guidelines, and was found to be applicable for routine quantitative analysis of Entecavir by RP-HPLC using UV detector in pharmaceutical dosage forms. The results of linearity, precision, accuracy and specificity, were proved, that does not exceed certain specified limits. The method provides selective quantification with no interference from other formulation excipients. The proposed method was highly sensitive, reproducible, reliable, robust and specific. Therefore, this method is a simple, rapid analysis may actually be more desirable than a more complicated and time-consuming process. The degradation studies at various stress conditions like thermal and hydrolytic, drug gets degraded at a temperature of 80 Â°c and refluxing with water at 70 Â°c for 24hours.
2. www.drugbank.ca/drugs/DB00709. [Last accessed on 10 Apr 2017].
3. Dimou E, Papadimitropoulos V, Hadziyannis SJ. The role of entecavir in the treatment of chronic hepatitis B. Ther Clin Risk Management 2007;3:1077-86.
4. Ching Lung Lai, Man-Fung Yuen. The saga of Entecavir. Hepatol Int 2009;3:421â€“24.
5. Lai CL, Rosmawati M, Lao J, Van Vlierberghe H, Anderson FH, Thomas N, et al. Entecavir is superior to lamivudine in reducing hepatitis B virus DNA in patients with chronic hepatitis B infection. Gastroenterology 2002;123:1831-8.
6. Kiran Kumar V, Raju NA. Spectrophotometric estimation of Entecavir in pharmaceutical dosage formulations. Biomed Pharmacol J 2008;1. http://biomedpharmajournal.org/?p=477
7. SM Malipatil, Bharath S, Athanikar, Mogal dipali. UV-spectrophotometric estimation of entecavir in tablet dosage form. Pharma Sci Monitoran Int J Pharm Sci 2012;3:56-67.
8. Dalmora SL, Sangoi Mda S, Nogueira DR, Da Silva LM. Validation of a stability-indicating RP-HPLC method for the determination of entecavir in tablet dosage form. J AOAC Int 2010;93:523-30.
9. Vijay Amirtharaj R, Vinay Kumar Ch, N Senthil Kumar. Development and validation of RP-HPLC for the estimation of entecavir in tablet dosage form. Int J Res Pharm Biomed Sci 2011;2:1033-40.
10. Delepee R, Got L, Causse X, Agrofoglio LA, Si Ahmed SN. Simultaneous determination of tenofovir, adefovir, lamivudine, entecavir in plasma. Poster No.576. 05A. viral hepatitisâ€“a) Hepatitis b)â€“Experimental; 2010. p. s215.
11. Jung BH, Naser L, Rezk, Bridges AS, Corbett AH, Angela D. Simultaneous determination of 17 antiretroviral drugs in human plasma for quantitative analysis with liquid chromatography-tandem mass spectrometry. Biomed Chromatogr 2007;21:1095â€“104.
12. Duxi Zhang,, Yunlin Fu, Jane P Gale, Anne F Aubry, Mark E Arnold. A sensitive method for the determination of entecavir at picogram per millilitre level in human plasma by solid phase extraction and high-pH LCâ€“MS/MS. J Pharm Biomed Anal 2009;49:1027â€“33.
13. Denny Yifei Liu, Erika Hess. Determination of entecavir in human plasma using Lc/Ms/Ms. Tandem Labs; 2010.
14. Rajeswari P Subrahmanyam, G Devala Rao, G Sudhakar Sai Babu. Novel spectrophotometric methods for the determination of entecavir in pharmaceutical dosage forms. Int J Pharma Bio Sci 2011;2:210-3.
15. E Shotton, K Ridgway. Physical Pharmaceutics. First Indian Edition; 2008. p. 326-8.
16. Napoleon. Pharmaceutical Titrimetric Analysis. First Edition; 2006. p. 4-44.
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.