Metabolic Test Substance Concentration (μg/mL) Exposure Harvest Activation Period Time 20, 40*, 80*, 120*, 160, 200 4 18 hours Absent μg/mL 20, 40*, 80*, 120*, 160, 200 4 hours 18 hours Present μg/mL *Cultures selected for metaphase analysis. Remarks Results The chromosomal aberration rates (16% without S9, 22.5% with S9) – were increased after treatment with 120 as compared to solvent μg/mL controls (statistically significant and biologically relevant). The number of cells carrying exchanges (8% and 12.5%, respectively) was also distinctly increased as compared to control levels. Furthermore, in the presence of metabolic activation, there was as dose-related increase in the number of aberrant cells. No increase in the frequencies of polyploid metaphases was found after treatment with the test item compared to controls. Test concentrations up to 200 were chosen as precipitation was μg/mL observed following treatment with 425 in the pre-experiment. μg/mL effects indicated by reduced cell numbers and/or mitotic indices of below 50% of control were observed in all tests at all concentrations. At 120 the number of cells was reduced to 11% (without S9) and μg/mL, 26% (with S9) of control values. Appropriate mutagens were used as positive controls. They induced statistically significant increases in cells with structural chromosome aberrations. The notified chemical was clastogenic to Chinese Hamster V79 cells treated in vitro under the conditions of the test RCC Ltd. (2003w) 7.10. Genotoxicity in vivo – Notified chemical. OECD TG 474 Mammalian Erythrocyte Micronucleus Test. EC Directive 2000/32/EC B.12 Mutagenicity Mammalian Erythrocyte Micronucleus Test. Species/Strain Mouse / NMRI Route of Administration Single administration/Oral Vehicle Corn oil. Remarks Method No significant protocol deviations. – Group Number and Sex Dose Sacrifice Time of Animals mg/kg bw Hours 1 (negative control) 5/sex 24 2 5/sex 500 24 3 5/sex 1000 24 4 5/sex 2000 24 5 5/sex 2000 48 6 (positive control) 5/sex 40 (CP) 24 CP=cyclophosphamide.
Metabolic Activation Test Substance Concentration (μg/mL) Exposure Period Harvest Time Absent 20, 40*, 80*, 120*, 160, 200 μg/mL 4 18 hours Present 20, 40*, 80*, 120*, 160, 200 μg/mL 4 hours 18 hours *Cultures selected for metaphase analysis. Remarks – Results The chromosomal aberration rates (16% without S9, 22.5% with S9) were increased after treatment with 120 μg/mL as compared to solvent controls (statistically significant and biologically relevant). The number of cells carrying exchanges (8% and 12.5%, respectively) was also distinctly increased as compared to control levels. Furthermore, in the presence of metabolic activation, there was as dose-related increase in the number of aberrant cells. No increase in the frequencies of polyploid metaphases was found after treatment with the test item compared to controls. Test concentrations up to 200 μg/mL were chosen as precipitation was observed following treatment with 425 μg/mL in the pre-experiment. effects indicated by reduced cell numbers and/or mitotic indices of below 50% of control were observed in all tests at all concentrations. At 120 μg/mL, the number of cells was reduced to 11% (without S9) and 26% (with S9) of control values. Appropriate mutagens were used as positive controls. They induced statistically significant increases in cells with structural chromosome aberrations. The notified chemical was clastogenic to Chinese Hamster V79 cells treated in vitro under the conditions of the test RCC Ltd. (2003w) 7.10. Genotoxicity – in vivo Notified chemical. OECD TG 474 Mammalian Erythrocyte Micronucleus Test. EC Directive 2000/32/EC B.12 Mutagenicity Mammalian Erythrocyte Micronucleus Test. Species/Strain Mouse / NMRI Route of Administration Single administration/Oral Vehicle Corn oil. Remarks – Method No significant protocol deviations. Group Number and Sex of Animals Dose mg/kg bw Sacrifice Time Hours 1 (negative control) 5/sex 24 2 5/sex 500 24 3 5/sex 1000 24 4 5/sex 2000 24 5 5/sex 2000 48 6 (positive control) 5/sex 40 (CP) 24 CP=cyclophosphamide.