GENOTOXIC IMPURITIES: AN IMPORTANT REGULATORY ASPECT
Keywords:Genotoxicity, Sources, Guidelines, Techniques for quantification, Control strategy, Limitations of present regulatory system to test genotoxicity
Genotoxins are agents/carriers such as chemical or radiation that can cause the damage to DNA or chromosomal structure, thereby causing mutations and the process are called as genotoxicity. Identification and understanding of genotoxins at a primary stage of drug development would enable us to prevent the potential damage that can be caused by these genotoxic agents. Various regulatory agencies such as International Council for Harmonization and EMEA, USFDA, European Pharmacopeia guidance, guidance for oncology products provide guidelines to limits the level of impurities in drug substances and drug products. Nowadays, conventional protocol of isolation, various spectral analysis high-performance liquid chromatography (LC), Fourier transform infrared to on-line analysis using modern, sophisticated hyphenated tools, like gas chromatography-mass spectroscopy, LC-MS so on, as well as modern software based in silico drug designs are extensively used by industry, research, and development areas and also there is tremendous increase in publications in the literature involving their use. Our review article focused on the various regulatory guidelines, application of hyphenated tools, and in silico techniques in genotoxic impurity and degradation product profiling of small molecules. A brief explanation is made on possible pitfalls in the experimentation and data interpretation. From this review, it concluded that there are various countries having their own regulatory agencies and regulatory guidelines for drug approvals, which may be followed by applying new chemical entities the new drug application title (NDA) in new drug application as well as there are various conventional to modern software based techniques to quantification of genotoxic impurities.
Savale SK. Genotoxicity of drugs: Introduction, prediction and evaluation. Asian J Biomater Res 2018;4:1-29.
Liu KT, Chen CH. Determination of impurities in pharmaceuticals: Why and how? In: Quality Management and Quality Control: New Trends and Developments. London, United Kingdom: IntechOpen; 2019.
Gosar A, Sayyed H, Shaikh T. Genotoxic impurities and its risk assessment in drug compounds. Drug Design Intellect Properties Int J 2018;2:227-32.
EFSA Scientific Committee. Scientific opinion on genotoxicity testing strategies applicable to food and feed safety assessment. EFSA J 2011;9:2379.
Shah SU. Importance of genotoxicity and S2A guidelines for genotoxicity testing for pharmaceuticals. IOSR J Pharm Biol Sci 2012;1:43-54.
Guideline IH. Impurities in new drug substances Q3A (R2). In: Proceedings of the International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use. Geneva, Switzerland: 2006. Available from: https://www.database. ich.org/sites/default/files/Q3A_R2__Guideline.pdf. [Last accessed on 2019 Oct 31].
International Conference on Harmonisation Guideline on Impurities in New Drug Products, Q3B(R2); 2006. Available from: https://www.ema. europa.eu/en/documents/scientific-guideline/ich-q-3-b-r2-impurities-new-drug-products-step-5en.pdf. [Last accessed on 2019 Oct 31].
International Conference on Harmonisation Guideline for Residual Solvents, Q3C(R4); 2019. Available from: https://www.ema.europa.eu/en/documents/scientific-guideline/international-conference-harmonisation-technical-requirements-registration-pharmaceuticals-human-use_en-33.pdf. [Last accessed on 2019 Nov 01].
European Medicines Agency. Guideline on the Specification Limits for Residues of Metal Catalysts or Metal Reagents, Doc. Ref. EMEA/ CHMP/SWP/4446/2000. Netherlands: European Medicines Agency; 2013. Available from: https://www.ema.europa.eu/documents/ scientific-guideline/guideline-specification-limits-residues-metal-catalysts-metal-reagents_en.pdf. [Last accessed on 2019 Oct 31].
European Medicines Agency. Guideline on the Limits of Genotoxic Impurities, CPMP/SWP/5199/02,EMEA/CHMP/QWP/251344/2006. European Medicines Agency; 2007. Available from: https://www.ema. europa.eu/documents/scientific-guideline/guideline-limits-genotoxic-impurities_en.pdf. [Last accessed on 2019 Oct 31].
Kroes R, Renwick AG, Cheeseman M, Kleiner J, Mangelsdorf I, Piersma A, et al. Structure-based thresholds of toxicological concern (TTC): Guidance for application to substances present at low levels in the diet. Food Chem Toxicol 2004;42:65-83.
Muller L, Mauthe RJ, Riley CM, Andino MM, Antonis DD, Beels C, et al. A rationale for determining, testing and controlling specific impurities in pharmaceuticals that possess potential for genotoxicity, regul. Toxicol Pharmacol 2006;44:198-211.
Center for Drug Evaluation and Research, Food and Drug Administration. Guidance (Draft) for Industry Genotoxic and Carcinogenic Impurities in Drug Substances and Products: Recommended Approaches. United States: Center for Drug Evaluation and Research, Food and Drug Administration; 2008.
Connelly JC. ICH guideline residual solvents. Counc pharmeur 1997;1:S1-68.
Callis CM, Bercu JP, DeVries KM, Dow LK, Robbins DK, Varie DL. Risk assessment of genotoxic impurities in marketed compounds administered over a short-term duration: Applications to oncology products and implications for impurity control limits. Org Process Res Dev 2010;14:986-92.
Leighton JK, Olejniczak K, Onodera H. ICH S9: Nonclinical evaluation of anticancer pharmaceuticals: A perspective from regulators on the development of the guideline. In: Global Approach in Safety Testing. New York: Springer; 2013. p. 283-98.
Venkatramani CJ, Sayah MA. Analytical strategies for genotoxic impurities in the pharmaceutical industry. Am Pharm Rev 2014;17:1-5.
Sultan S, Tengli A, Akhila G. Analytical assessment of genotoxic impurities in pharmaceuticals by gas chromatographic techniques. Res Rev 2019;1:22-32.
Mohamed SA, Sabita U, Rajendra S, Raman D. Genotoxicity: Mechanisms, testing guidelines and methods. Glob J Pharm Pharm Sci 2017;1:1-6.
Guideline IH. Assessment and Control of DNA Reactive (Mutagenic) Impurities in Pharmaceuticals To Limit Potential Carcinogenic RISK M7. Geneva: International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH); 2018. Available from: https://www.fda.gov/regulatory-information/search-fda-guidance-documents/m7r1-assessment-and-control-dna-reactive-mutagenic-impurities-pharmaceuticals-limit-potential [Last accessed on 2019 Nov 02].
Pierson DA, Olsen BA, Robbins DK, DeVries KM, Varie DL. Approaches to assessment, testing decisions, and analytical determination of genotoxic impurities in drug substances. Org Process Res Dev 2009;13:285-91.
Helmy R, Strickfuss S, Al-Sayah M, Hamilton S, Bu X, Lee C, et al. Quantification of Genotoxic Impurities in Active Pharmaceutical Ingredients. United Kingdom; Taylor & Francis Group; 2015. p. 293.
Luo L, Gu C, Li M, Zheng X, Zheng F. Determination of residual 4-nitrobenzaldehyde in chloramphenicol and its pharmaceutical formulation by HPLC with UV/Vis detection after derivatization with 3-nitrophenylhydrazine. J Pharm Biomed Anal 2018;156:307-12.
Jain M, Srivastava V, Kumar R, Dangi V, Hiriyanna SG, Kumar A, et al. Determination of five potential genotoxic impurities in dalfampridine using liquid chromatography. J Pharm Biomed Anal 2017;133:27-31.
Douša M, Doubský J, Srbek J. Utilization of photochemically induced fluorescence detection for HPLC determination of genotoxic impurities in the vortioxetine manufacturing process. J Chromatogr Sci 2016;54:1625-30.
Vundavilli A, Kumar J. Determination of hydroxylamine by HPLC, a mutagenic impurity in febuxostat drug substance. Int J Pharm Pharm Res 2018;12:318-30.
Huang Y, Lu H, Zhang F, Min C. Identification, isolation, characterization, and UHPLC quantification of potential genotoxic impurities in linagliptin. J Sep Sci 2018;41:3985-94.
Fu M, Lu Q, Hewitt E, Wang J. Ultra high performance liquid chromatography coupled with high resolution quantitation mass spectrometry method development and validation for determining genotoxic 2, 5-dichlorobenzoyl chloride in MLN9708 drug substance. J Pharm Biomed Anal 2014;89:233-9.
Hussain SZ, Maqbool K. GC-MS: Principle, technique and its application in food science. Int J Curr Sci 2014;13:116-26.
Parr MK, Joseph JF. NDMA impurity in valsartan and other pharmaceutical products: Analytical methods for the determination of N-nitrosamines. J Pharm Biomed Anal 2018;164:536-49.
Division of Pharmaceutical Analysis, Center for Drug Evaluation and Research. GC/MS Headspace Method for Detection of NDMA in Valsartan Drug Substance and Drug Products. United States: Division of Pharmaceutical Analysis, Center for Drug Evaluation and Research; 2019.
Reddy SR, Reddy KH, Kumar MN, Reddy PM, Reddy JV, Sharma HK. A validated GC-MS method for the determination of genotoxic impurities in divalproex sodium drug substance. J Chromatogr Sci 2018;57:101-7.
D’Souza AJ,Lokhande SR, Anvekar T. Development and validation of GCMS method for the detection and quantification of potential genotoxic impurity ethyl 4-bromobutyrate in tolvaptan tablets. Int J Res Appl Sci Eng Technol 2018;6:2460-5.
Zate PB, Kothari S, Lokhande MV. Confirmation and quantification of genotoxic impurity 2-dimethylaminoethyl chloride hydrochloride (DMC HCl) by GCMS in chlorpheniramine/chlorphenamine maleate. J Appl Chem 2017;10:21-6.
Maddala VL, Ray PC, Rao KN. A sensitive and selective GC-MS method for analysis of genotoxic impurities in dobutamine hydrochloride. Orient J Chem 2016;32:1685-90.
Gooty AR, Katreddi HR, Hunnur RK, Sharma HK, Masani NK. Simultaneous determination of genotoxic impurities in fudosteine drugs by GC-MS. J Chromatogr Sci 2016;54:1277-81.
Maddala VL, Ray PC, Venugopal K, Rao KM. A sensitive and selective GC-MS analysis of process related genotoxic impurities of nebivolol hydrochloride. Asian J Chem 2016;28:811-3.
Kakasaheb NA, Ramakrishna K, Srinivasarao V. Genotoxic impurity method development and validation by GCMS for the analysis of methyl methane sulfonate (MMS) in zidovudine drug substance. ACAIJ 2015;15:184-9.
Patrian B, Poiger T, Müller MD. Carbon Tetrachloride in Folpet Formulations by Headspace GC-MS. Available from: http://www. agroscope.ch.
Kakasaheb NA, Ramakrishna K, Srinivasarao V. Method development and validation by GC-MS for quantification of 1-chloroethyl cyclohexyl carbonate as a genotoxic impurity in candesartan cilexetil drug substance. Int J Pharm Pharm Sci 2014;6:370-2.
Akshatha HS, Gurupadayya BM. Application of liquid chromatography coupled with mass spectrometry in the impurity profiling of drug substances and products. Asian J Pharm Clin Res 2018;11:30-7.
Iliou K, Malenović A, Loukas YL, Dotsikas Y. Analysis of potential genotoxic impurities in rabeprazole active pharmaceutical ingredient via liquid chromatography-tandem mass spectrometry, following quality-by-design principles for method development. J Pharm Biomed Anal 2018;149:410-8.
Subbaiah N, Reddy GR, Kanyawar N, Gangrade M. A selective and sensitive method development and validation by Lc-Ms/Ms approach for trace level quantification of three potential genotoxic impurities in pantoprazole sodium drug substance. Rasayan J Chem 2017;10:1080-87.
Grigori K, Loukas YL, Malenović A, Samara V, Kalaskani A, Dimovasili E, et al. Chemometrically assisted development and validation of LC-MS/MS method for the analysis of potential genotoxic impurities in meropenem active pharmaceutical ingredient. J Pharm Biomed Anal 2017;145:307-14.
Srinivasu N, Ramachandran D. Determination of genotoxic impurity in atazanavir sulphate drug substance by LC-MS. J Pharm Biomed Anal 2017;132:156-8.
Rao LK, Devanna N, Reddy KV. Method development and validation study for quantitative determination of genotoxic impurity and its precursor in fluconazole sample by liquid chromatography-tandem mass spectrometry. Int J Pharm Pharm Sci 2016;8:84-9.
Jaishetty N, Palanisamy K, Maruthapillai A, Jaishetty R. Trace level quantification of the (−) 2-(2-amino-5-chlorophenyl)-4-cyclopropyl-1, 1, 1-trifluoro-3-butyn-2-ol genotoxic impurity in efavirenz drug substance and drug product using LC-MS/MS. Sci Pharm 2016;84:456-66.
Harigaya K, Yamada H, Horimoto S, Nishi H, Haginaka J. Sensitive quantitation of residual phenylhydrazine in antipyrine by LC-ICP-MS with iodo derivatization. Anal Sci 2014;30:845-50.
Narayana M, Chandrasekhar KB, Rao BM. Quantification of genotoxic impurity 2-butyl p-toluene-sulfonate at ppm level by LC-MS/MS in naproxen drug substance. Chem Sci 2014;3:929-36.
Amberg A, Czich A, Thybaud V. In silico/computational assessment for the evaluation of genotoxic impurities. Pharm Ind Pract Genotoxic Impurities 2014;1425:528-53.
Zhu Q, Li T, Wei X, Li J, Wang W. In silico and in vitro genotoxicity evaluation of descarboxyl levofloxacin, an impurity in levofloxacin. Drug Chem Toxicol 2014;37:311-5.
Emerce E, Cok I, Degim IT. Determination of the impurities in drug products containing montelukast and in silico/in vitro genotoxicological assessments of sulfoxide impurity. Toxicol Lett 2015;238:90-9.
Guideline IH. Assessment and Control of DNA Reactive (Mutagenic) Impurities in Pharmaceuticals to Limit Potential Carcinogenic RISK M7. Geneva: International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH); 2014.
Robinson DI. Control of genotoxic impurities in active pharmaceutical ingredients: A review and perspective. Org Process Res Dev 2010;14:946-59.
Singh S, Handa T, Narayanam M, Sahu A, Junwal M, Shah RP. A critical review on the use of modern sophisticated hyphenated tools in the characterization of impurities and degradation products. J Pharm Biomed Anal 2012;69:148-73.
Jena GB, Kaul CL, Ramarao P. Genotoxicity testing, regulatory requirement for drug discovery and development: Impact of ICH guidelines. Indian J Pharmacol 2002;34:86-99.
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