all doses in a dose-related manner. Desquamation occurred mainly following erythema; necrosis was observed in one rabbit receiving a dose of 2.15 mL/kg. Morphological findings: One rabbit in the 2.15 mL/kg dose group had pitted kidneys. LD50: > 4.64 mL/kg. Result: The test substance was of low dermal toxicity in rats. 9.1.3 Inhalation Toxicity (TM-694) (Tox Monitor Laboratories, 1994) Species/strain: rat/Sprague-Dawley. Number/sex of animals: 5/sex/dose group. Dose groups: 2.09 and 5.45 mg/L for 4 Observation period: 14 days. Method of administration: An aerosol was produced using an atomising system in a 100 L clear plastic chamber. Test method: OECD TG 403 Mortality: Complete mortality by day 3 in the 5.45 mg/L exposure group. Mortality of 20% by day 4 in the 2.09 mg/L exposure group. Clinical observations: 5.45 mg/L group: hypoactivity, wet fur in all animals, uncoordinated movement in 2 males until time of death. 2.09 mg/L group: uncoordinated movement in 1 male until time of death; piloerection and hypoactivity in 1 male which died and 1 male which did not; also hypoactivity only in 1 female. Morphological findings: 5.45 mg/L lungs red and mottled, g.i. tract distended. 2.09 mg/L group: as above in 2 males but less severe. LC50: Between 2.09 and 5.45 mg/L. Result: The notified chemical was of low acute inhalational toxicity in rats. 9.1.4 Skin Irritation (TM-592) (IBR-US, 1974) Species/strain: rabbit/albino.
Strains: E. coli WP2uvrA Metabolic activation: Microsomal fraction (S9) from Aroclor 1254-induced rat liver homogenate. Concentration range: 0, 10, 50, 100, 500, 1000 or 5000 g/plate (maximum of 2500 g/plate in the duplicate assay). Test method: OECD TG 471 Comment: Positive controls used were 2-anthramine to test the efficacy of the metabolic activation system and Positive controls demonstrated the sensitivity of the test system and negative controls were within historical limits. Result: The test substance was non mutagenic under the conditions of the test and no increase in the mean number of revertants above background occurred. 9.3.1.3 TM-599 (SRI International, 1993b) Strains: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 Metabolic activation: Microsomal fraction (S9) from Aroclor 1254-induced rat homogenate. Concentration range: 0, 10, 50, 100, 500, 1000 or 5000 g/plate (maximum of 2500 g/plate in the duplicate assay). Test method: OECD TG 471 Comment: Positive controls used were 2-anthramine to test the efficacy of the metabolic activation system, sodium and 2-nitrofluorene. Positive controls demonstrated the sensitivity of the test system and negative controls were within historical limits. Result: The test substance was non mutagenic under the conditions of the test and no increase in the mean number of revertants above background occurred. 9.3.1.4 TM-599D (SRI International, 1993c) Strains: E. coli WP2uvrA
demonstrated the test sensitivity. Negative control (vehicle) gave the expected response. Result: The test substance was non clastogenic under the conditions of the test and no increase in the frequency of micronucleated polychromatic erythrocytes occurred. 9.4 Overall Assessment of Toxicological Data Analogues of the notified chemical were of very low oral toxicity (LD50 > 5000 mg/kg, low acute dermal toxicity (LD50 > 4.64 mL/kg) and low acute inhalation toxicity (LC50 between 2.09 and 5.45 mg/L) in rats. Analogues of the notified chemical were slightly irritating to rabbit skin, not irritating to rabbit eyes and not skin sensitising in guinea pigs. The notified chemical exhibited diuretic effects in a 28-day oral repeated dose study in rats with a NOAEL of 50 mg/kg/day. Organ toxicity was limited to increased severity of hepatocyte microvacuolation at 500 mg/kg/day. A 90-day feeding study in rats was conducted on an analogue of the notified chemical. The NOAEL was 15 mg/kg/day on the basis of lower urine specific gravity in males, also an indicator of diuresis in the 28-day study with the notified chemical. Analogues of the notified chemical were not mutagenic in bacteria and were not clastogenic in mouse bone marrow cells in vivo. The test substance used to measure inhalation toxicity is determined to be a hazardous substance according to the NOHSC Approved Criteria for Classifying Hazardous Substances (NOHSC, 1999) and is assigned the risk phrase R20: Harmful by inhalation. Therefore, the notified chemical is assigned the risk phrase R20. 10. ASSESSMENT OF ENVIRONMENTAL EFFECTS A number of of the toxicity of the notified chemical or of a chemical analogue were provided, and the results of these test summarised below.
Easily visible translucent areas, details of iris slightly obscure 2 mild 50% to 75% 3 Opalescent areas, no details of iris visible, size of pupil barely discernible 3 moderate Greater than 75% 4 Opaque, iris invisible 4 severe CONJUNCTIVAE Redness Rating Chemosis Rating Discharge Rating Vessels normal Vessels definitely injected above normal More diffuse, deeper crimson red with individual vessels not easily discernible Diffuse beefy red 0 none 1 slight 2 mod. 3 severe No swelling Any swelling above normal Obvious swelling with partial eversion of lids Swelling with lids Swelling with lids to completely closed 0 none 1 slight 2 mild 3 mod. 4 severe No discharge Any amount different from normal Discharge with moistening of lids and adjacent hairs Discharge with moistening of lids and and considerable area around eye 0 none 1 slight 2 mod. 3 severe IRIS Values Rating Normal 0 none Folds above normal, congestion, swelling, circumcorneal injection, iris reacts to light 1 slight No reaction to light, haemorrhage, gross destruction 2 severe Draize, J. H., Woodward, G., Calvery, H. O. (1944) Methods for the Study of Irritation and Toxicity of Substances Applied Topically to the Skin and Mucous Membranes, J. Pharmacol. Exp. Ther. 82 : 377-390. Draize J. H. (1959) Appraisal of the Safety of Chemicals in Drugs and Cosmetics. Association of Food and Drug Officials of the US, 49 : 2-56.