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KMID : 1177720210150010045
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Journal of Alternatives to Animal Experiments
2021 Volume.15 No. 1 p.45 ~ p.55
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Reverse Mutation Evaluation for Eight Chemicals using Microplate Method
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Kim Myoung-Jun
Heo Seong-Moo
Nam Su-Hyun
Back Seng-Min
Park Seong-Hoon
Oh Jeong-Ja
Chung Young-Shin
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Abstract
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This study was conducted to establish the technology for bacterial reverse mutation test using 384-well microplates and liquid media instead of solid agar in order to evaluate the mutagenicity of test articles with limited amounts. The important advantage of this test method was considered that the usage of S9 fraction prepared from rat liver was smaller in comparison with the Ames test using solid agar. Five bacterial strains (S. typhimurium TA98, TA100, TA1535, TA1537 and E. coli WP2 uvrA) recommended in OECD TG 471 were used to evaluate 8 chemicals including 6 mutagens (Allyl chloride, 4-amino-2-nitrophenol, ethylene glycol dimethyl ether, benzo[α]pyrene, 7,12-dimethylbenzanthracene, 8-hydroxyquinoline) and 2 non-mutagens (dioctyl phthalate, sulfisoxazole). All experiments were performed using Ames MPFTM Penta II kits (Xenometrix, Switzerland). Three independent laboratories participated in the intra-laboratory and inter-laboratory studies. Results from this study were compared with reference mutagenicity data (NTP or published data). In results of this study, five out of six mutagens were evaluated to be positive, and the one mutagen (ethylene glycol dimethyl ether) to be negative. Two non-mutagens were determined to be negative in microplate tests. Both the intra-laboratory and the inter-laboratory study showed the 87.5% concordance in the overall results. It was concluded that a microplated reverse mutation test may be a compatible test method for the animal welfare as well as for the limited amount of substances.
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KEYWORD
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microplate Ames, bacterial reverse mutation, genotoxicity, concordance
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