Without metabolic activation for strain 156 to 5000 µg/plate E. coli With metabolic activation for strain: 156 to 5000 µg/plate E. coli Vehicle Water (for the notified chemical) Remarks - Method Concentration of the tested material was 30% Appropriate vehicle and positive controls were used. The negative controls were within normal limits and the positive controls sodium azide, aminopropylamino] acridine.2HCl, 2-aminoanthracene, Benzo(a)pyrene demonstrated the sensitivity of the test system. The mutagenicity study on strains was repeated without metabolic activation because of growth inhibition in the initial test. Metabolic Test Substance Concentration (µg/plate) Resulting in: Activation Cytotoxicity in Cytotoxicity in Main Precipitation Genotoxic Effect Preliminary Test Test Absent Test 1 > 78 for 78 for Not observed no S. S. typhimurium 2500 for typhimurium E. coli 5000 for E. coli Test 2 - 78 for Not observed no S. typhimurium Present Test 1 > 313 for 156 for Not observed no S. S. typhimurium 2500 for typhimurium E. coli 5000 for E. coli Test 2 - not performed - no Remarks - Results The observed cytotoxicity of the notified chemical at relatively low concentrations of the notified chemical is most likely due to the surfactant properties. The notified chemical is not mutagenic to bacteria under the conditions of the test. BML (1997) Genotoxicity in vitro – Notified chemical Study in compliance with Japanese regulatory standards for Toxicity testing of Pharmaceutical products and GLP standards. Species/Strain Chinese hamster Cell Type/Cell Line Lung fibroblasts (CHL/IU) cells Metabolic Activation System S9 fraction from Phenobarbital and 5,6-benzoflavone activated Rat liver at 5% Vehicle Saline Remarks - Method Concentration of the notified chemical in the test is stated to be ten times higher than in the table below. However, the dilution in the cell medium was not taken into account. Metabolic Test Substance Concentration (μg/mL) Exposure Harvest Activation Period Time Absent Test 1 4*; 8*; 12*; 16* 24h 24h Test 2 4*; 8*; 12*; 16* 48h 48h Test 3 7.8*; 15.6*; 31.3*; 62.5* 6h 24h
Without metabolic activation for E. coli strain 156 to 5000 µg/plate With metabolic activation for E. coli strain: 156 to 5000 µg/plate Vehicle Water (for the notified chemical) Remarks - Method Concentration of the tested material was 30% Appropriate vehicle and positive controls were used. The negative controls were within normal limits and the positive controls sodium azide, aminopropylamino] acridine.2HCl, 2-aminoanthracene, Benzo(a)pyrene demonstrated the sensitivity of the test system. The mutagenicity study on strains was repeated without metabolic activation because of growth inhibition in the initial test. Metabolic Activation Test Substance Concentration (µg/plate) Resulting in: Cytotoxicity in Preliminary Test Cytotoxicity in Main Test Precipitation Genotoxic Effect Absent Test 1 > 78 for S. typhimurium 5000 for E. coli 78 for S. typhimurium 2500 for E. coli Not observed no Test 2 - 78 for S. typhimurium Not observed no Present Test 1 > 313 for S. typhimurium 5000 for E. coli 156 for S. typhimurium 2500 for E. coli Not observed no Test 2 - not performed - no Remarks - Results The observed cytotoxicity of the notified chemical at relatively low concentrations of the notified chemical is most likely due to the surfactant properties. The notified chemical is not mutagenic to bacteria under the conditions of the test. BML (1997) Genotoxicity – in vitro Notified chemical Study in compliance with Japanese regulatory standards for Toxicity testing of Pharmaceutical products and GLP standards. Species/Strain Chinese hamster Cell Type/Cell Line Lung fibroblasts (CHL/IU) cells Metabolic Activation System S9 fraction from Phenobarbital and 5,6-benzoflavone activated Rat liver at 5% Vehicle Saline Remarks - Method Concentration of the notified chemical in the test is stated to be ten times higher than in the table below. However, the dilution in the cell medium was not taken into account. Metabolic Activation Test Substance Concentration (μg/mL) Exposure Period Harvest Time Absent Test 1 4*; 8*; 12*; 16* 24h 24h Test 2 4*; 8*; 12*; 16* 48h 48h Test 3 7.8*; 15.6*; 31.3*; 62.5* 6h 24h
EX130FR.docx