Positive controls: When with S9 mix 2-aminoanthracene for all strains. When without S9 mix 2-nitrofluorene (2-NF) for TA98; Methyl methanesulphonate (MMS) for TA100; 9-aminoacridine (9-AA) for TA1537; and N-ethyl-N-nitro-N-nitrosoguanidinne (ENNG) for TA1535 and WP2uvrA. Similar to OECD TG 471 & 472 Test method: Precipitation was observed at 312.5 g/plate and above. Comment: There was no toxicity to the bacteria under the test conditions. Under the conditions of the study, the notified chemical caused no substantial increases in revertant colony numbers over control counts at any concentration in either the presence or absence of the rat liver microsomal enzymes. All positive and negative controls responded appropriately. The notified chemical was considered to be non-mutagenic Result: under the conditions of the assay. 9.3.2 Chromosome aberration test in CHO cells (Murie, 2000) in vitro Chinese hamster (CHO) cells Liver fraction (S9 mix) from rats pretreated with Aroclor Metabolic activation: 1254. Two independent experiments were conducted in duplicate. DMSO was used as the vehicle. The experimental design Experimental design: and concentrations tested are tabulated below: Metabolic Experiment Test substance concentration (g/mL) Controls Activation No -S9 1 22 hour treatment: Positive: MMS. 0, 20, 39, 78, 156, 313, 625, 1250, 2 500 and 5 000. Negative: DMSO. 2 22 hour treatment (24 hour harvest): Positive: MMS. 0, 39, 78, 156, 313, 625 and 1 250. Negative: DMSO. 46 hour treatment (48 hour harvest): 0, 39, 78, 156, 313, 625 and 1 250.
Positive controls: When with S9 mix 2-aminoanthracene for all strains. When without S9 mix 2-nitrofluorene (2-NF) for TA98; Methyl methanesulphonate (MMS) for TA100; 9-aminoacridine (9-AA) for TA1537; and N-ethyl-N-nitro-N-nitrosoguanidinne (ENNG) for TA1535 and WP2uvrA. Test method: Similar to OECD TG 471 & 472 Comment: Precipitation was observed at 312.5 g/plate and above. There was no toxicity to the bacteria under the test conditions. Under the conditions of the study, the notified chemical caused no substantial increases in revertant colony numbers over control counts at any concentration in either the presence or absence of the rat liver microsomal enzymes. All positive and negative controls responded appropriately. Result: The notified chemical was considered to be non-mutagenic under the conditions of the assay. 9.3.2 Chromosome aberration test in CHO cells in vitro (Murie, 2000) Chinese hamster (CHO) cells Metabolic activation: Liver fraction (S9 mix) from rats pretreated with Aroclor 1254. Experimental design: Two independent experiments were conducted in duplicate. DMSO was used as the vehicle. The experimental design and concentrations tested are tabulated below: Metabolic Activation Experiment No Test substance concentration (g/mL) Controls -S9 1 22 hour treatment: 0, 20, 39, 78, 156, 313, 625, 1250, 2 500 and 5 000. Positive: MMS. Negative: DMSO. 2 22 hour treatment (24 hour harvest): 0, 39, 78, 156, 313, 625 and 1 250. 46 hour treatment (48 hour harvest): 0, 39, 78, 156, 313, 625 and 1 250. Positive: MMS. Negative: DMSO.
NA869FR.docx