Metabolic Test Substance Concentration (µg/plate) Resulting in: Activation Cytotoxicity in Cytotoxicity in Precipitation Genotoxic Effect Preliminary Test Main Test Absent Test 1 5,000 - - Negative ˃ Test 2 - 5,000 - Negative ˃ Present Test 1 5,000 - - Negative ˃ Test 2 - 5,000 - Negative ˃ Remarks - Results No toxic effects or no substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with the notified chemical at any dose level, neither in the presence nor absence of metabolic activation. In test 2, slight toxic effects were observed at 5,000 µg/plate in strains TA 1535 and TA 1537 with metabolic activation. The study authors concluded that there was no tendency of higher mutation rates with increasing concentrations of the notified chemical in the range below the generally acknowledged border of biological significance. The positive controls (sodium azide, 2-aminoanthracene, methyl methane sulfonate) showed a distinct increase in induced revertant colonies, confirming the efficacy of the test system. The notified chemical was not mutagenic to bacteria under the conditions of the test. RCC Ltd (2003t) Genotoxicity mammalian chromosome aberration test in vitro – Notified chemical (approximately 92% in purity) OECD TG 473 Mammalian Chromosome Aberration Test. In vitro EC Directive 2000/32/EC B.10 Mutagenicity - Mammalian In vitro Chromosome Aberration Test. Species/Strain Chinese Hamster Cell Type/Cell Line V79 Metabolic Activation System S9 fraction induced rat liver Phenobarbital/β-naphthoflavone Vehicle Deionised water Remarks - Method No significant protocol deviations. Metabolic Test Substance Concentration Exposure Harvest (μg/mL) Activation Period (hrs) Time (hrs) Absent Test 1 10, 20*, 40*, 60*, 80, 100 4 18 Test 2 5, 10*, 20*, 40*, 60, 80 18 18 Test 3 20*, 40, 60, 80 28 28 Present Test 1 10, 20, 40*, 60*, 80*, 100 4 18 Test 2 10*, 20*, 40*, 60, 80, 100 4 28 *Cultures selected for metaphase analysis.
Metabolic Activation Test Substance Concentration (µg/plate) Resulting in: Cytotoxicity in Preliminary Test Cytotoxicity in Main Test Precipitation Genotoxic Effect Absent Test 1 ˃ 5,000 - - Negative Test 2 - ˃ 5,000 - Negative Present Test 1 ˃ 5,000 - - Negative Test 2 - ˃ 5,000 - Negative Remarks - Results No toxic effects or no substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with the notified chemical at any dose level, neither in the presence nor absence of metabolic activation. In test 2, slight toxic effects were observed at 5,000 µg/plate in strains TA 1535 and TA 1537 with metabolic activation. The study authors concluded that there was no tendency of higher mutation rates with increasing concentrations of the notified chemical in the range below the generally acknowledged border of biological significance. The positive controls (sodium azide, 2-aminoanthracene, methyl methane sulfonate) showed a distinct increase in induced revertant colonies, confirming the efficacy of the test system. The notified chemical was not mutagenic to bacteria under the conditions of the test. RCC Ltd (2003t) Genotoxicity – in vitro mammalian chromosome aberration test Notified chemical (approximately 92% in purity) OECD TG 473 In vitro Mammalian Chromosome Aberration Test. EC Directive 2000/32/EC B.10 Mutagenicity - In vitro Mammalian Chromosome Aberration Test. Species/Strain Chinese Hamster Cell Type/Cell Line V79 Metabolic Activation System S9 fraction Phenobarbital/β-naphthoflavone induced rat liver Vehicle Deionised water Remarks - Method No significant protocol deviations. Metabolic Activation Test Substance Concentration (μg/mL) Exposure Period (hrs) Harvest Time (hrs) Absent Test 1 10, 20*, 40*, 60*, 80, 100 4 18 Test 2 5, 10*, 20*, 40*, 60, 80 18 18 Test 3 20*, 40, 60, 80 28 28 Present Test 1 10, 20, 40*, 60*, 80*, 100 4 18 Test 2 10*, 20*, 40*, 60, 80, 100 4 28 *Cultures selected for metaphase analysis.