IN-SILICO TOXICITY ASSESSMENT AND TRACE LEVEL QUANTIFICATION OF VERATRYL CHLORIDE A POTENTIAL GENOTOXIC IMPURITY IN IVABRADINE HYDROCHLORIDE USING LC-MS/MS
Abstract
A sensitive and selective liquid chromatography-mass spectrometry (LC-MS/MS) method using multiple reaction monitoring (MRM) mode was developed and validated for trace analysis of 4-(chloromethyl)-1, 2-dimethoxybenzene (Veratryl chloride) a potential genotoxic impurity in Ivabradine hydrochloride (IVB). Chromatographic separation was performed on a Poroshell 120EC C18 (50 × 3.0 mm, 2.7 μm) column using a mixture of 10 mM ammonium formate and acetonitrile in isocratic elution mode at a 0.25 mL/min flow rate. A simple pre-column derivatization with di-ethylamine was employed for the derivatization of the veratryl chloride. The developed LC-MS/MS method was linear and accurate in the 1.5–10.0 ppm concentration range with r2 ˃ 0.999 and percent recoveries greater than 90%. The developed method was precise with RSD (%) of not more than 4.5%. In-silico genotoxicity and carcinogenicity potential of veratryl chloride was assessed using ICH M7 principles found to be positive. The developed method can identify and quantify veratryl chloride in IVB, hence can be applied by quality control labs of pharmaceutical industries for trace quantification.
Keywords:
LC-MS/MS, MRM, Genotoxic impurity, In-silico toxicity, Method validation, Pre-column derivatizationDOI
https://doi.org/10.25004/IJPSDR.2023.150413References
Giordani A, Kobel W and Gally HU. Overall impact of the regulatory requirements for genotoxic impurities on the drug development process. Eur J Pharm Sci. 2011; 43:1-15.
ICH M7(R1): Assessment and control of DNA reactive (Mutagenic) Impurities in pharmaceuticals to limit potential carcinogenic risk. 2017. https://database.ich.org/sites/default/files/M7_R1_Guideline.pdf
Yancy CW, Jessup M, Bozkurt B, Butler J, Casey Jr DE, Colvin MM, Drazner MH, Filippatos G, Fonarow GC, Givertz MM, Hollenberg SM, Lindenfeld J, Masoudi FA, McBride PE, Peterson PN, Stevenson LW, Westlake C. 2017 ACC/AHA/HFSA Focused update of the 2013 ACCF/AHA guideline for the management of heart failure: A report of the American College of Cardiology/American Heart Association Task Force on Clinical Practice Guidelines and the Heart Failure Society of America. Circulation. 2017; 136:e137-e161.
Tardif JC, Ford I, Tendera M, Bourassa MG, Fox K. Efficacy of Ivabradine, a new selective I(f) inhibitor, compared with atenolol in patients with chronic stable angina. Eur Heart J. 2005; 26:2529-2536.
Thollon C, Cambarrat C, Vian J, Prost JF, Peglion JL, Vilaine JP. Electrophysiological effects of S 16257, a novel sino-atrial node modulator, on rabbit and guinea-pig cardiac preparations. comparison with UL-FS 49. Br J Pharmacol. 1994; 112: 37-42.
Sulfi S, Tmimmis AD. Ivabradine – The first selective sinus node I(f) channel inhibitor in the treatment of stable angina. Int J Clin Pract. 2006; 60: 222-228.
Muller L, Mauthe RJ, Riley CM, Andino MM, Antonis DD, Beels C, George JDe, De Knaep AGM, Ellison D, Fagerland JA, Frank R, Fritschel B, Galloway S, Harpur E, Humfrey CDN, Jacks AS, Jagota N, Mackinnon J, Mohan Genes DK, Donovan MRO, Smith MD, Vudathala G, Yotti L. A rationale for determining, testing, and controlling specific impurities in pharmaceuticals that possess potential for genotoxicity. Regul Toxicol Pharmacol. 2006; 44:198-211.
Snodin DJ. Genotoxic Impurities: From Structural Alerts to Qualification. Org Process Res Dev. 2010; 14: 960-976.
Ravi Kiran P, Punna Rao R, Syam Kumar MB, Sumanth M, Chandra Sekhar KVG. In silico toxicity assessment and trace level quantification of two genotoxic impurities in silodosin using capillary gas chromatography. J Anal Sci Technol. 2023; 14:15.
Ravi Kiran P. In silico toxicity studies and trace level quantification of genotoxic impurities in pharmaceutical drug substances. Department of Chemistry, Birla Institute of Technology and Science, Pilani, India (2022).
Sunitha S, Srinivasan BP. Development and Validation of RP-HPLC Method for the Estimation of Ivabradine Hydrochloride in Tablets. Indian J Pharm Sci.2010; 72: 667-671.
Ranjha NM, Majeed A, Hussain I, Rasool MF. Quantitative Determination and Validation of Ivabradine-HCl in Pharmaceutical Formulation and Rabbit Plasma by High Performance Liquid Chromatography Method. Curr Pharm Anal. 2017; 13:446-451.
Pundkar SA, Vyas VP. Analytical method development and validation for estimation of ivabradine hydrochloride by using RP-HPLC. Asian J Res Bio Pharm Sci. 2019; 7:29-43.
Tomic J, Djajic N, Agbaba D, Otasevic B, Malenovic A, Protic A. Robust optimization of gradient RP HPLC method for simultaneous determination of ivabradine and its eleven related impurities by AQbD approach. Acta Chromatogr. 2021; 34:1-11.
Tomic J, Ivkovic B, Olijacic S, Nikolic K, Naljuric N, Protic A, Agbaba D. Chemometricall assisted RP-HPLC method development for efficient separation of ivabradine and its eleven impurities. Acta Chromatogr. 2020; 32:53-63.
ICH Q2A(R1). Validation of Analytical Procedures: Text and Methodology. 2005. https://database.ich.org/sites/default/files/Q2%28R1%29%20Guideline.pdf
US FDA. Analytical Procedures and Methods Validation for Drugs and Biologics. 2015. https://www.fda.gov/files/drugs/published/Analytical-Procedures-and-Methods-Validation-for-Drugs-and-Biologics.pdf
Jerry M, Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Wiley (1992)
Liu DQ, Sun M, Kord AS. Analytical control of genotoxic impurities in the pazopanib hydrochloride manufacturing process. J Pharm Biomed Anal. 2009; 50:144-150.
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