METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF AMPRENAVIR IN HUMAN PLASMA BY LIQUID CHROMATOGRAPHY–ELECTROSPRAY IONIZATION–TANDEM MASS SPECTROMETRY
Objective: The main aim of the present study was to develop a sensitive liquid chromatography–electrospray ionization–tandem mass spectrometric technique for the quantitation of amprenavir in human plasma.
Methods: Chromatographic separation was achieved on a reversed-phase Symmetry C18 (50 mm×4.6 mm, 3.5 μm) column with isocratic elution by acetonitrile and 0.1% v/v formic acid in the ratio of 90:10 v/v as mobile phase. Chromatographic peaks were resolved with 0.7 ml/min flow rate. Drug was extracted with ethyl acetate solvent by liquid–liquid extraction method. Monitoring of transition of m/z 506.2 and 71.0 for amprenavir and 628 and 421 for methyl-indinavir was made on multiple reaction monitoring.
Results: Calibration curve of amprenavir was linear over 1–600 ng/ml concentration range with regression coefficient (r2) value of >0.99. The % relative standard deviation values were <8.5% for interday and intraday precision and accuracy. The method has excellent recovery, and the percentage recovery values of lower quality control (QC), median QC, and higher QC samples were 101.86%, 102.8%, and 99.28%, respectively.
Conclusion: The drug was stable for more time at variable stability conditions, and method was successfully applicable to regular analysis of amprenavir in biological matrices.
2. Wittayanarakul K, Hannongbua S, Feig M. Accurate prediction of protonation state as a prerequisite for reliable MM-PB(GB)SA binding free energy calculations of HIV-1 protease inhibitors. J Comput Chem 2008;29:673-85.
3. Dandache S, Sévigny G, Yelle J, Stranix BR, Parkin N, Schapiro JM, et al. In vitro antiviral activity and cross-resistance profile of PL-100, a novel protease inhibitor of human immunodeficiency virus Type 1. Antimicrob Agents Chemother 2007;51:4036-43.
4. Sadler BM, Hanson CD, Chittick GE, Symonds WT, Roskell NS. Safety and pharmacokinetics of amprenavir (141W94), a human immunodeficiency virus (HIV) Type 1 protease inhibitor, following oral administration of single doses to HIV-infected adults. Antimicrob Agents Chemother 1999;43:1686-92.
5. Agenerase (amprenavir) Capsules. Full Prescribing Information. Section Dosage and Administration. US Food and Drug Administration. GlaxoSmithKline and Vertex Pharmaceuticals Inc. Available from: . [Last retrieved on 2015 Nov 29].
6. Rajitha K, Prasanna NL, Naveen R, Ranjith CH, Kumar AA. A rapid RP-HPLC method development and validation for the quantitative estimation of indinavir in capsules. Int J Pharm Pharm Sci 2014;6:453-6.
7. Padmini T, Satyanarayana L. Spectrophotometric determination of amprenavir by complex formation in bulk drug and formulation samples. Int J Pharm Clin Res 2017;9:521-4.
8. Chi J, Jayewardene AL, Stone JA, Motoya T, Aweeka FT. Simultaneous determination of five HIV protease inhibitors nelfinavir, indinavir, ritonavir, saquinavir and amprenavir in human plasma by LC/MS/MS. J Pharm Biomed Anal 2002;30:675-84.
9. Koehn J, Ding Y, Freeling J, Duan J, Ho RJ. A simple, efficient, and sensitive method for simultaneous detection of anti-HIV drugs atazanavir, ritonavir, and tenofovir by use of liquid chromatography-tandem mass spectrometry. Antimicrob Agents Chemother 2015;59:6682-8.
10. Das Mishra T, Kurani H, Singhal P, Shrivastav PS. Simultaneous quantitation of HIV-protease inhibitors ritonavir, lopinavir and indinavir in human plasma by UPLC-ESI-MS-MS. J Chromatogr Sci 2012;50:625-35.
11. US FDA. Guidance for Industry Bioanalytical Method Validation, Food and Drug Administration, Center for Drug Evaluation and Research (CDER). Rockville, Maryland: USA; 2001.
12. Tijare LK, Rangari NT, Mahajan UN. Areview on bioanalytical method development and validation. Asian J Pharm Clin Res 2016;9:6-10.
13. Liliya L, Dmytro K, Oleksandra O, Taras P, Mariya D. A HPLC-MS/MS method development and validation for the simultaneous determination of bisoprolol and enalapril in the present of enalaprilat in human plasma. Int J Appl Pharm 2018;10:31-40.
14. Swathi P, Vidyadhara S, Sasidhar RL, Chakravarthi KK. Method development and validation for the estimation of entecavir in bulk and pharmaceutical dosage forms by RP-HPLC. Int J Curr Pharm Res 2017;9:107-11.
15. Ravi Y, Rajkamal B. An improved LC-MS/MS method development and validation for the determination of trandolapril and verapamil in human plasma. Int J Pharm Pharm Sci 2018;11:91-5.
16. ICH: Q2B. Harmonized Tripartite Guideline, Validation of Analytical Procedure: Methodology, IFPMA. Geneva: Proceedings of the International Conference on Harmonization; 1996.
17. ICH: Q2 (R1). Validation of Analytical Procedures: Text and Methodology. International Conference on Harmonisation; 2005.
18. Lakshmi PR, Prahlad P, Ravindra N, Rao MV. A rapid and sensitive liquid chromatography mass spectrometry/mass spectrometry method for estimation of pioglitazone, keto pioglitazone and hydroxy pioglitazone in human plasma. Asian J Pharm Clin Res 2017;10:120-8.
19. Hepsebah NJ, Kumar AA. Bioanalytical method development and validation of ibrutinib in biological matrices by LC-MS/MS. Int J Pharm Pharm Sci 2018;11:22-6.
This work is licensed under a Creative Commons Attribution 4.0 International License.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.