Vehicle Dimethyl sulfoxide Remarks - Method No significant protocol variations. Metabolic Test Substance Concentration (µg/plate) Resulting in: Activation Cytotoxicity in Cytotoxicity in Precipitation Genotoxic Effect Preliminary Test Main Test Absent Test 1 >2500 - Negative Test 2 - Negative Present Test 1 >2500 >2500 - Negative Test 2 >2500 - Negative Remarks - Results No mutagenic effects were observed with or without metabolic activation. Results with positive controls demonstrated the validity of the test system. The notified chemical was not mutagenic to bacteria under the conditions of the test. JAI Research Foundation (2009j) Genotoxicity in – Notified chemical OECD TG 476 In vitro Mammalian Cell Gene Mutation Test. Species/Strain Chinese hamster ovary Cell Type/Cell Line Cell line CHO-K1 Metabolic Activation System Extract of rat liver (treated with Aroclor 1254) called S9 fraction was used. The S9 fraction was buffered and supplemented with the essential co-factors NADP, KCl and glucose-6-phosphate to form S9 mix. β Vehicle Dimethyl sulfoxide Remarks - Method No significant protocol variations. Concentrations for testing were selected on the basis of a preliminary cytotoxicity test. S9 was used at 2% in Test 2 with metabolic activation, and at 1% in the remaining tests. Metabolic Test Substance Concentration (μg/mL) Exposure Expression Selection Activation Period Time Time Absent Test 1 31.25, 62.5*, 125*, 250*, 500* 4 h 24 h 48 h Test 2 31.25, 62.5*, 125*, 250*, 500* 23 h 24 h 48 h Present Test 1 31.25, 62.5*, 125*, 250*, 500* 4 h 24 h 48 h Test 2 31.25, 62.5*, 125*, 250*, 500* 4 h 24 h 48 h *Cultures selected for metaphase analysis. Metabolic Test Substance Concentration (µg/mL) Resulting in: Activation Cytotoxicity in Cytotoxicity in Precipitation Genotoxic Effect Preliminary Test Main Test >500 Absent Test 1 >500 >500 Negative
Vehicle Dimethyl sulfoxide Remarks - Method No significant protocol variations. Metabolic Activation Test Substance Concentration (µg/plate) Resulting in: Cytotoxicity in Preliminary Test Cytotoxicity in Main Test Precipitation Genotoxic Effect Absent Test 1 >2500 - Negative Test 2 - Negative Present Test 1 >2500 >2500 - Negative Test 2 >2500 - Negative Remarks - Results No mutagenic effects were observed with or without metabolic activation. Results with positive controls demonstrated the validity of the test system. The notified chemical was not mutagenic to bacteria under the conditions of the test. JAI Research Foundation (2009j) Genotoxicity – in Notified chemical OECD TG 476 In vitro Mammalian Cell Gene Mutation Test. Species/Strain Chinese hamster ovary Cell Type/Cell Line Cell line CHO-K1 Metabolic Activation System Extract of rat liver (treated with Aroclor 1254) called S9 fraction was used. The S9 fraction was buffered and supplemented with the essential co-factors β NADP, KCl and glucose-6-phosphate to form S9 mix. Vehicle Dimethyl sulfoxide Remarks - Method No significant protocol variations. Concentrations for testing were selected on the basis of a preliminary cytotoxicity test. S9 was used at 2% in Test 2 with metabolic activation, and at 1% in the remaining tests. Metabolic Activation Test Substance Concentration (μg/mL) Exposure Period Expression Time Selection Time Absent Test 1 31.25, 62.5*, 125*, 250*, 500* 4 h 24 h 48 h Test 2 31.25, 62.5*, 125*, 250*, 500* 23 h 24 h 48 h Present Test 1 31.25, 62.5*, 125*, 250*, 500* 4 h 24 h 48 h Test 2 31.25, 62.5*, 125*, 250*, 500* 4 h 24 h 48 h *Cultures selected for metaphase analysis. Metabolic Activation Test Substance Concentration (µg/mL) Resulting in: Cytotoxicity in Preliminary Test Cytotoxicity in Main Test Precipitation Genotoxic Effect Absent >500 Test 1 >500 >500 Negative
STD1346FR.docx