{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T10:00:09Z","timestamp":1790762409261,"version":"4.1.0"},"reference-count":36,"publisher":"Wiley","issue":"8-9","license":[{"start":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T00:00:00Z","timestamp":1790726400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T00:00:00Z","timestamp":1790726400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100022416","name":"Center for Drug Evaluation and Research","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100022416","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Environ and Mol Mutagen"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Nitrosamine drug impurities, including both small\u2010molecule nitrosamines and nitrosamine drug substance\u2010related impurities (NDSRIs), are members of a cohort of concern for pharmaceutical safety assessment. As the toxicity of most nitrosamine impurities requires metabolic activation by cytochrome P450 enzymes, this study used human HepaRG cells, which have metabolic activity similar to that of human hepatic cells, to evaluate methods for nitrosamine safety assessment. The genotoxicity of four small\u2010molecule nitrosamines,\n                    <jats:italic>N\u2010<\/jats:italic>\n                    bis(2,2\u2010diethoxyethyl) nitrous amide (BDEA),\n                    <jats:italic>N<\/jats:italic>\n                    \u2010nitrosodiphenylamine,\n                    <jats:italic>N,N<\/jats:italic>\n                    \u2010nitroso\u2010\n                    <jats:italic>N<\/jats:italic>\n                    \u2010ethylaniline (NEPA), and 1\u2010methyl\u20104\u2010nitrosopiperazine (MNP), and five NDSRIs,\n                    <jats:italic>N<\/jats:italic>\n                    \u2010nitroso\u2010bumetanide,\n                    <jats:italic>N<\/jats:italic>\n                    \u2010nitroso\u2010ciprofloxacin,\n                    <jats:italic>N<\/jats:italic>\n                    \u2010nitroso\u2010dabigatran etexilate,\n                    <jats:italic>N<\/jats:italic>\n                    \u2010nitroso\u2010desmethyl\u2010diphenhydramine, and\n                    <jats:italic>N<\/jats:italic>\n                    \u2010nitroso\u2010sertraline, was evaluated in two\u2010dimensional (2D)\/attached and three\u2010dimensional (3D)\/spheroid HepaRG cell cultures using two high\u2010throughput genetic toxicity assays, the CometChip DNA damage assay and the flow\u2010cytometric micronucleus (MN) assay. These test substances exhibited a range of responses in other genotoxicity assays, including strong positive, negative, and inconclusive results. Following a 24\u2010h treatment, two NDSRIs,\n                    <jats:italic>N<\/jats:italic>\n                    \u2010nitroso\u2010desmethyl\u2010diphenhydramine and\n                    <jats:italic>N<\/jats:italic>\n                    \u2010nitroso\u2010sertraline, and two small\u2010molecule nitrosamines, BDEA and MNP, induced DNA damage, with the small\u2010molecule nitrosamines positive only in 3D HepaRG spheroids. MN formation was detected only in spheroids exposed to\n                    <jats:italic>N<\/jats:italic>\n                    \u2010nitroso\u2010desmethyl\u2010diphenhydramine. Benchmark concentration (BMC) values derived from the DNA damage data were lower for the genotoxic NDSRIs compared to the genotoxic small\u2010molecule nitrosamines, which correlated with higher cytotoxicity for the NDSRIs. These results show that, while HepaRG spheroids were a more sensitive model for the hazard identification of small\u2010molecule nitrosamines, the responses produced by NDSRIs were more complex, with NDSRIs tested producing similar levels of DNA damage in 2D and 3D cultures and inducing MN formation only in 3D spheroids.\n                  <\/jats:p>","DOI":"10.1002\/em.70082","type":"journal-article","created":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T09:02:37Z","timestamp":1790758957000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["<scp>DNA<\/scp>\n                    Damage and Micronucleus Induction by Small\u2010Molecule Nitrosamine Drug Impurities and Nitrosamine Drug Substance\u2010Related Impurities in\n                    <scp>HepaRG<\/scp>\n                    Cells"],"prefix":"10.1002","volume":"67","author":[{"given":"Hannah S.","family":"Xu","sequence":"first","affiliation":[{"name":"Division of Genetic and Molecular Toxicology National Center for Toxicological Research, U.S. Food and Drug Administration  Jefferson Arkansas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ji\u2010Eun","family":"Seo","sequence":"additional","affiliation":[{"name":"Division of Genetic and Molecular Toxicology National Center for Toxicological Research, U.S. Food and Drug Administration  Jefferson Arkansas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xilin","family":"Li","sequence":"additional","affiliation":[{"name":"Division of Genetic and Molecular Toxicology National Center for Toxicological Research, U.S. Food and Drug Administration  Jefferson Arkansas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3501-9014","authenticated-orcid":false,"given":"Nan","family":"Mei","sequence":"additional","affiliation":[{"name":"Division of Genetic and Molecular Toxicology National Center for Toxicological Research, U.S. Food and Drug Administration  Jefferson Arkansas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aisar H.","family":"Atrakchi","sequence":"additional","affiliation":[{"name":"Center for Drug Evaluation and Research, U.S. Food and Drug Administration  Silver Spring Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karen L.","family":"Davis Bruno","sequence":"additional","affiliation":[{"name":"Center for Drug Evaluation and Research, U.S. Food and Drug Administration  Silver Spring Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sruthi T.","family":"King","sequence":"additional","affiliation":[{"name":"Center for Drug Evaluation and Research, U.S. Food and Drug Administration  Silver Spring Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert 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USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5951-2156","authenticated-orcid":false,"given":"Xiaoqing","family":"Guo","sequence":"additional","affiliation":[{"name":"Division of Genetic and Molecular Toxicology National Center for Toxicological Research, U.S. Food and Drug Administration  Jefferson Arkansas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,30]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1093\/mutage\/geag011","article-title":"Nitrosamine Ames Data Review and Method Development: Proceedings of a US FDA\/HESI Workshop","volume":"41","author":"Atrakchi A.","year":"2026","journal-title":"Mutagenesis"},{"key":"e_1_2_11_3_1","doi-asserted-by":"crossref","first-page":"105835","DOI":"10.1016\/j.yrtph.2025.105835","article-title":"HESI GTTC Ring Trial: Concordance Between Ames and Rodent Carcinogenicity 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