7. TOXICOLOGICAL INVESTIGATIONS Endpoint and Result Assessment Conclusion Rat, acute oral LD50 > 2500 mg/kg bw, low toxicity Rat, dermal* LD50 > 2000 mg/kg bw, low toxicity Rat, acute inhalation Not available Rabbit, skin irritation Moderately irritating Rabbit, eye irritation* Severely irritating Mouse, skin sensitisation – LLNA** Evidence of sensitisation Rat, repeat dose oral toxicity – 28 days. NOEL = 200 mg/kg bw/day Genotoxicity – bacterial reverse mutation Non mutagenic Genotoxicity – in chromosomal aberration Genotoxic Genotoxicity – in vivo micronucleus assay Non genotoxic *Analogue chemical **Local lymph node assay The analogue chemical contains reactive functional groups, are likely to contribute to toxicity, e.g., irritation and sensitisation. 7.1. Acute toxicity – oral Notified chemical OECD TG 423 Acute Oral Toxicity – Acute Toxic Class Method. Species/Strain Rat/Sprague-Dawley CD strain rat Vehicle Arachis oil BP Remarks - Method There were no significant deviations from the protocol. Group Number and Sex of Animals Dose mg/kg bw Mortality 1 3 F 2000 0 2 3 F 2000 0 LD50 > 2500 mg/kg bw Signs of Toxicity There were no signs of systemic toxicity. All animals showed expected gains in bodyweight over the study period. Effects in Organs No abnormalities were noted at necropsy. The notified chemical is of low toxicity via the oral SafePharm Laboratories (2004) 7.2. Acute toxicity – dermal Analogue chemical (90%) OECD TG 402 Acute Dermal Toxicity – Limit Test. Species/Strain Rat/Strain unknown Vehicle Unknown Type of dressing Unknown Remarks - Method Unknown Group Number and Sex of Animals Dose mg/kg bw Mortality
Concentration mg/L Number of Fish Mortality Nominal Actual 1 h 24 h 48 h 72 h 96 h 0 0 7 0 0 0 0 0 100 90-94 7 0 0 0 0 0 LC50 > 100 mg/L at 96 hours. NOEC 100 mg/L at 96 hours. Remarks – Results In the control and in the 100 mg/L test, all fish survived until the end of the test and no visible abnormalities were observed. No remarkable observations were made concerning the appearance of the test medium. The test medium was a clear solution throughout the whole test duration. The notified chemical was not harmful to fish at a concentration of 100 mg/L. RCC 8.2.2. Acute toxicity to aquatic invertebrates Notified chemical OECD TG 202 Daphnia sp. Acute Immobilisation Test and Reproduction Test – Static. EC Directive 92/69/EEC C.2 Acute Toxicity for Daphnia – Static. Species Daphnia Exposure Period 48 Auxiliary Solvent None Water Hardness 250 mg Analytical Monitoring HPLC Remarks - Method The test medium was prepared by dissolving 30.2 mg of test item completely in 300 mL of test water by intense stirring for 15 minutes at room temperature. No significant deviations from the test guidelines were reported. Concentration mg/L Number of D. magna Number Immobilised Nominal Actual 24 h 48 h 0 0 20 0 0 100 98-112 20 0 0 LC50 > 100 mg/L at 48 hours NOEC 100 mg/L at 48 hours Remarks - Results The test medium was a clear solution throughout the duration of the test. The notified chemical was not harmful to daphnids at a concentration of 100 mg/L. RCC (2006f) 8.2.3. Algal growth inhibition test Notified chemical OECD TG 201 Alga, Growth Inhibition Test. EC Directive 92/69/EEC C.3 Algal Inhibition Test. Species Scenedesmus subspicatus Exposure Period 72 hours