Genotoxicity Reverse Mutation Assay (Bourner et al., 1993) Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100 Strains: 0.5, 5, 50, 500, 5 000 g/plate Concentration range: EC Method B 14 (similar to OECD TG 471) Test method: the notified chemical was toxic at the top dose as Comments: judged by lack of effect on microcolony formation; precipitation above 500 g/plate was predicted to obscure scoring of mutants and some precipitation was seen at this dose; positive control mutagens, which established the sensitivity of the assay were: 2-aminoanthracene: +S9; 2-nitrofluorene: TA 98 sodium azide: TA 1535 and TA 100 –S9; 9-aminoacridine: TA 1537 –S9 the notified chemical was not mutagenic at Result: up to 500 in bacteria in either the g/plate absence or presence of metabolic activation provided by Aroclor 1254-induced SD rat liver S9 fraction Cytogenetic Study in Human Lymphocytes (Beard et al., 1993) In vitro phytohaemagglutinin-stimulated human Cells: peripheral lymphocytes from female donors 500 for approximately 19 or 43 hours in Doses: g/mL the absence of metabolic activation provided Aroclor 1254-induced rat liver S9 fraction (S9) and in its presence for 3 hours similar to OECD TG 473 Test method: the dose level was chosen to reduce the mitotic Comments: index to 40 80% of controls; negative controls – were within the expected limits; the positive control substances were ethylmethane sulphonate (500 or cyclophosphamide (12.5 g/mL) g/mL) and demonstrated the sensitivity of the assay
Genotoxicity Salmonella typhimurium Reverse Mutation Assay (Bourner et al., 1993) Strains: TA 1535, TA 1537, TA 98, TA 100 Concentration range: 0.5, 5, 50, 500, 5 000 g/plate Test method: EC Method B 14 (similar to OECD TG 471) Comments: the notified chemical was toxic at the top dose as judged by lack of effect on microcolony formation; precipitation above 500 g/plate was predicted to obscure scoring of mutants and some precipitation was seen at this dose; positive control mutagens, which established the sensitivity of the assay were: 2-aminoanthracene: +S9; 2-nitrofluorene: TA 98 –S9; sodium azide: TA 1535 and TA 100 9-aminoacridine: TA 1537 –S9 Result: the notified chemical was not mutagenic at up to 500 g/plate in bacteria in either the absence or presence of metabolic activation provided by Aroclor 1254-induced SD rat liver S9 fraction In vitro Cytogenetic Study in Human Lymphocytes (Beard et al., 1993) Cells: phytohaemagglutinin-stimulated human peripheral lymphocytes from female donors Doses: 500 g/mL for approximately 19 or 43 hours in the absence of metabolic activation provided Aroclor 1254-induced rat liver S9 fraction (S9) and in its presence for 3 hours Test method: similar to OECD TG 473 Comments: the dose level was chosen to reduce the mitotic index to 40 – 80% of controls; negative controls were within the expected limits; the positive control substances were ethylmethane sulphonate (500 g/mL) or cyclophosphamide (12.5 g/mL) and demonstrated the sensitivity of the assay