Species/strain: Rat/Sprague-Dawley (CD) Number/sex of animals: 5/sex per group Observation period: 15 days Method of administration: Oral (gavage) doses at 2 000, 3 200, 5 000 and 6 400 mg/kg. Test method: OECD TG 401 Mortality: 1 male at 3 200 mg/kg; 1 male and 3 females at 5 000 mg/kg; and 3 at 6 400 mg/kg. Clinical observations: Piloerection, hunched posture, red brown stained muzzle, ungroomed appearance and walking on toes were seen in most animals at all dose levels. Increased salivation, waddling/unsteady gait, lethargy, sensitivity to handling, respiratory distress, abnormal faeces, cold extremities, body tremors, increased lacrimation, prostration, thin appearance, pallid extremities, protruding eyes and partially close eyelids were observed at one or more dosages. All symptoms reversed by day 14 except piloerection, which was still evident on day 15 in males at 5000 mg/kg. Morphological findings: A generalised congestion was observed in either all or the majority of organs and tissues of the dead animals. No macroscopic abnormalities were found in surviving animals. Comment: All deaths (1 male was killed in extremis) occurred within 3 days of dosing. LD50: 5 537 mg/kg (males), 4 938 mg/kg (females), 5 313 mg/kg (combined Result: The notified chemical was of very low acute oral toxicity in rats. 9.1.2 Dermal Toxicity (McRae, 1998a) Species/strain: Rat/Sprague-Dawley(CD) Number/sex of animals: 5/sex Observation period: 15 days
Method of administration: A dermal application (2 000 mg/kg in corn oil) under occlusive dressing for 24 hours. Test method: OECD TG 402 Mortality: None Clinical observations: None Morphological findings: None Comment: 3 female animals had either no bodyweight gain or low bodyweight gain. LD50: > 2 000 mg/kg Result: The notified chemical was of low dermal toxicity in rats. 9.1.3 Inhalation Toxicity Not provided. 9.1.4 Skin Irritation (Parcell, 1998b) Species/strain: Rabbit/New Zealand White Number/sex of animals: 6 Observation period: 72 hours Method of administration: A single application of the notified chemical (0.5 g) in corn oil under a semi-occlusive dressing for 4 hours. Test method: OECD TG 404 Draize scores: Draize scores for erythema and oedema were zero for all animals at 1, 24, 48 and 72 hours after dosing. Result: The notified chemical was not irritating to the skin of rabbits. 9.1.5 Eye Irritation (Parcell, 1998a) Species/strain: Rabbit/New Zealand Number/sex of animals: 6 females
Observation period: 72 hours Method of administration: A volume of 0.1 mL of the notified chemical was placed into the everted lid of one eye of each animal. The other eye served as control. Test method: OECD TG 405 Draize scores: Time instillation Animal 1 hour 1 day 2 days 3 days Conjunctiva r c r c r c r c 1 2 0 2 0 0 0 0 0 2 1 0 1 0 0 0 0 0 3 1 0 1 0 0 0 0 0 4 1 0 1 0 0 0 0 0 5 1 0 1 0 0 0 0 0 6 1 0 0 0 0 0 0 0 1 see Attachment 1 for Draize scales r = redness c = chemosis Other Draize scores: Draize scores for cornea (density and area) and iris lesion in all 6 animals were zero during the study. Comment: Discharge from conjunctivae was not recorded. Result: The notified chemical was a slight irritant to the eyes of rabbits. 9.1.6 Skin Sensitisation (Coleman, 1998) Species/strain: Guinea pigs/albino Dunkin- Hartley Number of animals: 20 (test group); 20 males (test vehicle group); 10 males (positive control group); 10 males (vehicle control group). Induction: Intradermal injection (Day 1) 3 pairs of intradermal injections (0.1 mL) were made on shaved shoulder area of each animal: Test Freund’s Complete Adjuvant (FCA) with water (1:1 v/v);
-S9 First test Treatment/recovery time =3/18 hours 6.25, 12.5, 25, 50, 100*, 200*, 400* and 800 g/mL Positive: mitomycin C (0.8 g/mL) Negative: DMSO Positive: mitomycin C (0.4 g/mL) Negative: DMSO Second test Treatment time=21 hours 50, 100, 150, 200*, 300*, 400*, 500* and 600 g/mL +S9 First test Treatment/recovery time =3/18 hours 6.25, 12.5, 25, 50, 100*, 200*, 400* and 800* g/mL Positive: cyclophosphamide (20 g/mL) Negative: DMSO Second test Treatment/recovery time =3/18 50, 100, 200*, 400*, 500, 600, 700 and 800* g/mL DMSO – dimethylsulphoxide * cultures selected for metaphase analysis Test method: OECD TG 473 Comment: The notified chemical showed toxic at 400 and 800 g/mL without S9 and 800 g/mL with S9 in the first test, and showed toxic at 600 g/mL without S9 in the second test. Precipitates were observed at the 235.5 g/mL in culture medium when DMSO was used as the solvent. The notified chemical did not show indication of inducing chromosomal aberrations in the absence or presence of metabolic activation in this in vitro cytogenetic test system. Both positive control compounds caused large, statistically significant increases in the proportion of aberrant cells. Result: The notified chemical was non clastogenic under the conditions of the test. 9.3.3 Mammalian Cell Gene Mutation Test in vitro in the Mouse (Adams, 1998) Cells: Mouse lymphoma L5178Y cells Metabolic activation system: Liver fraction (S9 mix) from rats pretreated with Aroclor 1254.