Comment: None. LD50: >2 000 mg/kg Result: The notified chemical was of very low acute oral toxicity in rats. 9.1.2 Dermal Toxicity (Edgar, 1999a) Species/strain: Rat/Sprague Dawley Number/sex of animals: 5/sex Observation period: 14 days Method of administration: A dermal dose of 2 000 mg/kg was applied under an occlusive dressing for 24 hours. Test method: OECD TG 402, limit test Mortality: No deaths were recorded. Clinical observations: Red discharge from the nose was observed on Day 1. Morphological findings: No abnormalities were noted at necropsy. Comment: None, LD50: > 2 000 mg/kg Result: The notified chemical was of low dermal toxicity in rats. 9.1.3 Inhalation Toxicity An inhalation toxicity study was not provided. 9.1.4 Skin Irritation (Edgar, 1999b) Species/strain: Rabbit/New Zealand White Number/sex of animals: 3 females. Observation period: 7 days Method of administration: A volume of 0.5 mL was applied under a semi-occlusive dressing for 4 hours. Test method: OECD TG 404
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.
+S9 1 6 treatment: 0, 20, 39, 78, 156, 313, 625, 1250, 2 500 and 5 000. Positive: CPH. Negative: DMSO. 2 6 hours treatment (24 hour harvest): 0, 39, 78, 625, 1 250 and 2 500. 6 treatment (48 hour harvest): 0, 39, 78, 625, 1 250 and 2 500. Positive: CPH. Negative: DMSO. - cultures selected for metaphase analysis DMSO: Dimethyl sulphoxide CPH: cyclophosphamide Test method: OECD TG 473 Comment: Precipitation increased gradually from 78 g/mL. Cytotoxicity was observed at 156 and 625 g/mL and above with and without S9 mix, respectively in the first test, and at 625 and 156 g/mL and above with and without S9 mix, respectively in the second test. The test substance did not induce any significant or increases in the frequency of cells with aberrations in either experiment. All positive and negative controls responded appropriately. Result: The notified chemical was considered to be non-clastogenic under the conditions of the study. 9.3.3 Micronucleus Assay in the Bone Marrow Cells of the Mouse (Watson & Innes, 2000) Species/strain: Mice/CD-1 Number and sex of animals: Test group: 10/sex, Vehicle control 5/sex, Positive control group: 5 males. Dosing: 2 000 mg/kg at 0 and 24 hours. Sampling: 48 hours. Vehicle: maize oil. Positive control: CPH. Method of administration: Oral (gavage). Test method: OECD TG 474