Appendix B: Toxicological Investigations All studies were conducted using a mixture of the notified chemicals. Acute toxicity oral – Notified chemicals (isomers present at: 32.1% and 60.3%) trans- cis- OECD TG 423 Acute Oral Toxicity Acute Toxic Class Method. – Species/Strain Rat/ Wistar (RccHanTM:WIST) Vehicle Arachis oil BP (300 mg/kg bw) or none (2,000 mg/kg bw). Remarks - Method No significant protocol deviations. GLP Compliance. Group Number and Sex Dose Mortality of Animals mg/kg bw 1 3F 300 0/3 2 3F 2,000 0/3 3 3F 2,000 0/3 LD50 > 2,000 mg/kg bw Signs of Toxicity Clinical observations seen in animals treated at 2,000 mg/kg bw included piloerection (3/6 animals) and hunched posture (6/6 animals). One of these animals was seen to additionally exhibit ptosis, ataxia, lethargy and laboured, noisy and gasping respiration. This animal was still showing signs at the end of the observation period. Signs of toxicity were observed in all other animals from 1 hour to 4 hours post dosing only. No signs of systemic toxicity were noted in any animals treated at 300 mg/kg bw. Effects in Organs No macroscopic abnormalities were seen at necropsy in any of the test animals. Remarks - Results All test animals showed body weight gains during the study period, except for one animal treated at 2,000 mg/kg bw, which showed weight loss during the first week, but gained weight during the second week of observation. This was the same animal which showed additional signs of toxicity throughout the study period. The mixture of notified chemicals is of low toxicity via the oral route. Harlan (2013d) Acute toxicity dermal – Notified chemicals (isomers present at: 32.1% and 60.3%) trans- cis- OECD TG 402 Acute Dermal Toxicity Limit Test. – Species/Strain Rat/Wistar (RccHanTM:WIST) Vehicle None. Type of dressing Semi-occlusive. Remarks - Method No significant protocol deviations. GLP Compliance. Group Number and Sex Dose Mortality of Animals mg/kg bw 1 5 per sex 2,000 0/10
Appendix B: Toxicological Investigations All studies were conducted using a mixture of the notified chemicals. Acute toxicity – oral Notified chemicals (isomers present at: trans- 32.1% and cis- 60.3%) OECD TG 423 Acute Oral Toxicity – Acute Toxic Class Method. Species/Strain Rat/ Wistar (RccHanTM:WIST) Vehicle Arachis oil BP (300 mg/kg bw) or none (2,000 mg/kg bw). Remarks - Method No significant protocol deviations. GLP Compliance. Group Number and Sex of Animals Dose mg/kg bw Mortality 1 3F 300 0/3 2 3F 2,000 0/3 3 3F 2,000 0/3 LD50 > 2,000 mg/kg bw Signs of Toxicity Clinical observations seen in animals treated at 2,000 mg/kg bw included piloerection (3/6 animals) and hunched posture (6/6 animals). One of these animals was seen to additionally exhibit ptosis, ataxia, lethargy and laboured, noisy and gasping respiration. This animal was still showing signs at the end of the observation period. Signs of toxicity were observed in all other animals from 1 hour to 4 hours post dosing only. No signs of systemic toxicity were noted in any animals treated at 300 mg/kg bw. Effects in Organs No macroscopic abnormalities were seen at necropsy in any of the test animals. Remarks - Results All test animals showed body weight gains during the study period, except for one animal treated at 2,000 mg/kg bw, which showed weight loss during the first week, but gained weight during the second week of observation. This was the same animal which showed additional signs of toxicity throughout the study period. The mixture of notified chemicals is of low toxicity via the oral route. Harlan (2013d) Acute toxicity – dermal Notified chemicals (isomers present at: trans- 32.1% and cis- 60.3%) OECD TG 402 Acute Dermal Toxicity – Limit Test. Species/Strain Rat/Wistar (RccHanTM:WIST) Vehicle None. Type of dressing Semi-occlusive. Remarks - Method No significant protocol deviations. GLP Compliance. Group Number and Sex of Animals Dose mg/kg bw Mortality 1 5 per sex 2,000 0/10