DEVELOPMENT AND VALIDATION OF A STABILITY INDICATING HPLC METHOD FOR THE DETERMINATION OF NILOTINIB HYDROCHLORIDE IN BULK AND PHARMACEUTICAL DOSAGE FORM
Objective: To develop a rapid, accurate, linear, sensitive and stability indicating RP-HPLC method for the determination of nilotinib in bulk and pharmaceutical dosage forms in the presence of its four related substances.
Methods: The RP-HPLC method was developed for the chromatographic separation of nilotinib and its impurities by using waters Xterra RP-18 (150*4.6 mm, 3.5 Âµm) column with a mobile phase combination of 10 mM ammonium formate with pH-3.5 and acetonitrile in gradient mode. An injection volume of 20 Âµl. Flow rate was 1.0 ml/min and detection was carried a wavelength of 250 nm. The method was validated as per ICH guidelines.
Results: The retention time for nilotinib and its four impurities were found to be 4.37, 7.40, 8.96, 10.21 and 10.87 min respectively. The linear regression analysis data for the calibration plots showed the good linear relationship in the concentration range of 0.04-3.0 ppm for the nilotinib impurities. The % recovery of nilotinib impurities was found to be 96.8-99.4% in the linearity range. The detection limit (LOD) values were about 0.014, 0.016, 0.005 and 0.03 ppm respectively and the quantification limit (LOQ) values were 0.042, 0.048, 0.014 and 0.09 ppm respectively. The % degradation at various stress conditions like acid, alkaline, oxidative, thermal and photolytic stress was found to be 8.92, 18.35,5.63, 0.88 and 3.89 respectively.
Conclusion: The RP-HPLC method compatible with LC-MS was developed for the analysis of nilotinib and its four impurities. It was validated as per the ICH guidelines and found to be linear, robust, precise, accurate, sensitive, stability indicating and can be used for routine as well as stability analysis of capsule dosage forms as well as for drug substance.
2. Weisberg E, Manley P, Mestan J, Cowan JS, Ray A, Griffin JD. AMN107 (nilotinib): a novel and selective inhibitor of BCR-ABL. Br J Cancer 2006;94:1765-9.
3. Marek D, Marie-Christine W, Daniel BL, Thomas F, Heidemarie B. High-performance liquid chromatography with ultraviolet detection and protein precipitation as a way of quantitative determination of nilotinib with and without an internal standard. J Liq Chromatogr Relat Technol 2012;35:2503-10.
4. Pursche S, Ottmann OG, Ehninger G, Schleyer E. High-performance liquid chromatography method with ultraviolet detection for the quantification of the BCR-ABL inhibitor nilotinib (AMN107) in plasma, urine, culture medium and cell preparations. J Chromatogr B: Anal Technol Biomed Life Sci 2007;852:208-16.
5. Haouala A, Zanolari B, Rochat B, Montemurro M, Zaman K, Duchosal MA, et al. Therapeutic drug monitoring of the new targeted anticancer agents imatinib, nilotinib, dasatinib, sunitinib, sorafenib and lapatinib by LC tandem mass spectrometry. J Chromatogr B: Anal Technol Biomed Life Sci 2007;877:1982-96.
6. Silvia DF, Antonio DA, Francesca DM, Elisa P, Lorena B, Marco S, et al. New HPLCâ€“MS method for the simultaneous quantification of the antileukemia drugs imatinib, dasatinib and nilotinib in human plasma. J Chromatogr B: Anal Technol Biomed Life Sci 2007;877:1721-6.
7. Masatomo M, Naoto Takahashi, Ken IS. High-performance liquid chromatography with solid-phase extraction for the quantitative determination of nilotinib in human plasma. Biomed Chromatogr 2010;24:789-93.
8. Yuki M, Yamakawa Y, Uchida T, Nambu T, Kawaguchi T, Hamada A, et al. High-performance liquid chromatographic assay for the determination of nilotinib in human plasma. Biol Pharm Bull 2011;34:1126-8.
9. Prenen H, Guetens G, de Boeck G, Debiec RM, Manly P, SchÃ¶ffski P, et al. Cellular uptake of the tyrosine kinase inhibitors imatinib and AMN107 in gastrointestinal stromal tumor cell lines. Pharmacology 2006;77:11-6.
10. Satyanarayana L, Naidu SV, Narasimha Rao M, Suma Latha R. The estimation of nilotinib in capsule dosage form by RP-HPLC. Asian J Pharm Technol 2011;1:82-4.
11. Sudhakar BK, Venkataramanna M, Mohanareddy C, Narayanareddy P, Latha J. A validated stability indicative UPLC method for nilotinib hydrochloride for the determination of process related and degradation impurities. J Chromatogr Sci 2014;52:880-5.
12. Harika M, Kumar GS. Development and validation of method for the determination of nilotinib by RP-HPLC in bulk and pharmaceutical dosage forms. Int Res J Pharm 2012;3:161-4.
13. ICH guidance on method validation; Validation of analytical procedures Text and Methodology Q2; 2005.
14. ICH harmonized tripartite guideline; Validation of analytical methods Definitions and Terminology EMEA; 1994.
15. ICH harmonized tripartite guideline; Stability testing of new drug substances and products Q1A; 2003.
16. Blessy M, Ruchi DP, Prajesh NP, Agarwal YK. Development of forced degradation and stability indicating studies of drugs. J Pharm Anal 2014;4:159-65.
17. ICH harmonized tripartite guideline; Photostability testing of new drug substances and products Q1B; 1996.
18. Sowjanya G, Annapurna MM, Sriram AV. Development and validation of a stability indicating RP-HPLC method for the determination of nilotinib (A tyrosine kinase inhibitor). Indo Am J Pharm Res 2013;3:4541-51.