{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T17:34:13Z","timestamp":1791999253660,"version":"4.4.0"},"reference-count":69,"publisher":"Oxford University Press (OUP)","issue":"2","license":[{"start":{"date-parts":[[2023,12,19]],"date-time":"2023-12-19T00:00:00Z","timestamp":1702944000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,3,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The robust control of genotoxic N-nitrosamine (NA) impurities is an important safety consideration for the pharmaceutical industry, especially considering recent drug product withdrawals. NAs belong to the \u2018cohort of concern\u2019 list of genotoxic impurities (ICH M7) because of the mutagenic and carcinogenic potency of this chemical class. In addition, regulatory concerns exist regarding the capacity of the Ames test to predict the carcinogenic potential of NAs because of historically discordant results. The reasons postulated to explain these discordant data generally point to aspects of Ames test study design. These include vehicle solvent choice, liver S9 species, bacterial strain, compound concentration, and use of pre-incubation versus plate incorporation methods. Many of these concerns have their roots in historical data generated prior to the harmonization of Ames test guidelines. Therefore, we investigated various Ames test assay parameters and used qualitative analysis and quantitative benchmark dose modelling to identify which combinations provided the most sensitive conditions in terms of mutagenic potency. Two alkyl-nitrosamines, N-nitrosodimethylamine (NDMA) and N-nitrosodiethylamine (NDEA) were studied. NDMA and NDEA mutagenicity was readily detected in the Ames test and key assay parameters were identified that contributed to assay sensitivity rankings. The pre-incubation method (30-min incubation), appropriate vehicle (water or methanol), and hamster-induced liver S9, alongside Salmonella typhimurium strains TA100 and TA1535 and Escherichia coli strain WP2uvrA(pKM101) provide the most sensitive combination of assay parameters in terms of NDMA and NDEA mutagenic potency in the Ames test. Using these parameters and further quantitative benchmark dose modelling, we show that N-nitrosomethylethylamine (NMEA) is positive in Ames test and therefore should no longer be considered a historically discordant NA. The results presented herein define a sensitive Ames test design that can be deployed for the assessment of NAs to support robust impurity qualifications.<\/jats:p>","DOI":"10.1093\/mutage\/gead033","type":"journal-article","created":{"date-parts":[[2023,12,15]],"date-time":"2023-12-15T15:25:17Z","timestamp":1702653917000},"page":"78-95","source":"Crossref","is-referenced-by-count":50,"title":["Ames test study designs for nitrosamine mutagenicity testing: qualitative and quantitative analysis of key assay parameters"],"prefix":"10.1093","volume":"39","author":[{"given":"Dean N","family":"Thomas","sequence":"first","affiliation":[{"name":"GSK Research & Development, Genetic Toxicology and Photosafety , Stevenage SG1 2NY , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John W","family":"Wills","sequence":"additional","affiliation":[{"name":"GSK Research & Development, Genetic Toxicology and Photosafety , Stevenage SG1 2NY , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helen","family":"Tracey","sequence":"additional","affiliation":[{"name":"GSK Research & Development, Genetic Toxicology and Photosafety , Stevenage SG1 2NY , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandy J","family":"Baldwin","sequence":"additional","affiliation":[{"name":"GSK Research & Development, Genetic Toxicology and Photosafety , Stevenage SG1 2NY , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark","family":"Burman","sequence":"additional","affiliation":[{"name":"GSK Research & Development, Genetic Toxicology and Photosafety , Stevenage SG1 2NY , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abbie N","family":"Williams","sequence":"additional","affiliation":[{"name":"GSK Research & Development, Genetic Toxicology and Photosafety , Stevenage SG1 2NY , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Danielle S G","family":"Harte","sequence":"additional","affiliation":[{"name":"GSK Research & Development, Genetic Toxicology and Photosafety , Stevenage SG1 2NY , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruby A","family":"Buckley","sequence":"additional","affiliation":[{"name":"GSK Research & Development, Genetic Toxicology and Photosafety , Stevenage SG1 2NY , United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1533-7664","authenticated-orcid":false,"given":"Anthony M","family":"Lynch","sequence":"additional","affiliation":[{"name":"GSK Research & Development, Genetic Toxicology and Photosafety , Stevenage SG1 2NY , United Kingdom"},{"name":"School of Medicine, Swansea University , Singleton Park, Swansea SA2 8PP , United 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