SIMULTANEOUS REVERSE-PHASE ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATION OF GRAZOPREVIR AND ELBASVIR

Authors

  • Madhavi S Department of pharmaceutical sciences, University College of Pharmaceutical Sciences, Acharya Nagarjuna University, Guntur, Andhra Pradesh, India
  • Prameela Rani A Department of pharmaceutical sciences, University College of Pharmaceutical Sciences, Acharya Nagarjuna University, Guntur, Andhra Pradesh, India

DOI:

https://doi.org/10.22159/ajpcr.2018.v11i4.21721

Keywords:

Grazoprevir, Elbasvir, Ultra performance liquid chromatography, Development, Validation, Forced degradation

Abstract

 Objective: The objective of this study was to develop and validate rapid, specific, sensitive, and precise reverse-phase ultra performance liquid chromatography (RP-UPLC) method for the quantitative determination of grazoprevir and elbasvir, as there are no official monograph and no analytical method by UPLC.

Methods: Chromatographic separation was achieved on a Waters Acquity UPLC HSS C18 (2.1 mm × 100 mm, 1.8 micron) column with a 45:55 (v/v) mixture of 0.1% orthophosphoric acid (pH 2.8) and acetonitrile as a mobile phase, thermostated at 30°C with a short run time of 3.0 min.

Results: The retention times were 0.73 and 1.29 min for grazoprevir and elbasvir, respectively. Quantification is achieved with TUV detection at 254 nm over the concentration range of 25–150 μg/ml for grazoprevir and for elbasvir 12.5–75 μg/ml, with a correlation coefficient of 0.999 and 0.999, respectively. The developed method was validated according to the International Conference on Harmonization (ICH) guidelines with respect to linearity, accuracy, precision, specificity, and robustness. Forced degradation study was extended out under acidic, alkaline, oxidative, photolytic, and thermal conditions to demonstrate the stability-indicating capability of the developed UPLC method. The degradation products were well resolved from the main peak, thus proved the stability-indicating power of the method. The results of the analysis were validated statistically.

Conclusion: The method is precise, accurate, linear, robust, and fast. The short retention time allows the analysis of a large number of samples in a short period of time and, therefore, should be cost-effective for routine analysis in the pharmaceutical industry.

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References

WHO Fact Sheet, 2016. Available from: http://www.who.int/mediacentre/Factsheets/fs164/en. [Last accessed on 2017 Jul 15].

Baid-Agrawal S, Pascual M, Moradpour D, Somasundaram R, Muche M. Hepatitis C virus infection and kidney transplantation in 2014: What’s new? Am J Transplant 2014;14:2206-20.

Zepatier (Elbasvir and Grazoprevir). Tablets for Oral Use. Whitehouse Station, NJ, USA: Merck & Co.; 2016.

Papudesu C, Kottilil S, Bagch S. Elbasvir/grazoprevir for treatment of chronic hepatitis C virus infection. Hepatol Int 2017;11:152-60.

Polamreddy P, Vishwakarma V, Gundla R. A review on anti-HCV agents targeting active site and allosteric sites of non-structural protein 5b [NS5B]. Int J Pharm Pharm Sci 2016;8:1-18.

Bell AM, Wagner JL, Barber KE. Elbasvir/grazoprevir: A review of the latest agent in the fight against hepatitis C. Int J Hepatol 2016;2016:1-7.

Liua H, Xua H, Song W. Validated UPLC/MS/MS assay for quantitative bioanalysis of elbasvir in rat plasma and application to pharmacokinetic study. J Chromatogr B 2016;1015:150-6.

Potluri H, Battula SR, Yeturu S. Picogram level quantification of grazoprevir and elbasvir with deuterated internal standards in human plasma samples by LC–ESI-MS/MS. Indian J Pharm Educ Res 2016;50:612-9.

Akram N, Umamahesh M, Ramachari T. A new validated RP-HPLC method for the determination of elbasvir and grazoprevir in its bulk and pharmaceutical dosage forms. Int J Chem Pharm Anal 2017;4:1-11.

Vancha AR, Naresh D, Sowjanya1 P, Kumar GV. Analytical method development and validation for elbasvir and grazoprevir in combine pharmaceutical dosage forms by RP-HPLC. Int J Res Dev Pharm Life Sci 2016;4:94-101.

International Conference on Harmonization. ICH Q2 (R1) Guideline on Validation of Analytical Procedures: Text and Methodology Yokohama, Japan; 2005.

Ponnuri RN, Pragallapati P, Mastanamma SK, Ravindra N, Mandava VB. Development and validation of a stability indicating reverse phase high performance liquid chromatography method for simultaneous determination of clindamycin, metronidazole, and clotrimazole in pharmaceutical combined dosage forms. Asian J Pharm Clin Res 2017;10:111-7.

Reddy GN, Prasad VV, Maharana PK. Development and validation of a stability indicating UPLC method for determination of erlotinib in pharmaceutical formulations. Pharm Chem 2012;4:2288-97.

Sunnis MC, Rajput AP. Development and validation of a new stability indicating analytical method for the determination of related components of brimonidine tartrate in drug substances and drug product using apples. Int J Pharm Pharm Sci 2011;3:145-50.

Balan P, Kannappan N. Development and validation of stability-indicating RP-UPLC method for simultaneous estimation of thiocolchicoside and aceclofenac in combined dosage form. Int Curr Pharm J 2014;3:296-300.

Yanamandra R, Vadla CS, Puppala UM, Patro B. Development and validation of a rapid RPUPLC method for the simultaneous estimation of bambuterol hydrochloride and montelukast sodium from tablets. Indian J Pharm Sci 2012;74:116-21.

Vasanth AD, Rajkamal B. A UPLC-MS/MS method development and validation for the estimation of pomalidomide from human plasma. Int J App Pharm 2017;9:37-43.

Published

01-04-2018

How to Cite

S, M., and P. Rani A. “SIMULTANEOUS REVERSE-PHASE ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATION OF GRAZOPREVIR AND ELBASVIR”. Asian Journal of Pharmaceutical and Clinical Research, vol. 11, no. 4, Apr. 2018, pp. 100-4, doi:10.22159/ajpcr.2018.v11i4.21721.

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