using Bacteria. Plate incorporation procedure and Pre incubation procedure Species/Strain TA1535, TA1537, TA98 and TA100 S. typhimurium: WP2uvrA E. coli: Metabolic Activation System Phenobarbital and -naphthoflavone induced rat liver microsomal fraction (S9) in a) With metabolic activation: 31.6 5,000 µg/plate – Main Test b) Without metabolic activation: 31.6 5,000 µg/plate – Vehicle DMSO Remarks - Method Main Test 1 and Test 2 used plate incorporation procedure and pre incubation procedure, respectively. Positive controls: 10 µg/plate sodium azide (NaN3) for TA100 and TA1535 10 µg/plate 4-nitro-o-phenylene-diamine (4-NOPD) for TA98 40 µg/plate 4-nitro-o-phenylene-diamine (4-NOPD) for TA1537 1 µL/plate methyl methane sulfonate (MMS) for WP2uvrA 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 > 5,000 > 5,000 Negative Test 2 - > 5,000 > 5,000 Negative Present Test 1 > 5,000 > 5,000 > 5,000 Negative Test 2 - > 5,000 > 5,000 Negative Remarks - Results No precipitation and cytotoxicity of the test substance were observed up to the highest concentration tested (5,000 µg/plate) in both the absence and the presence of the metabolic activation. The positive controls produced distinct increase of revertants indicative of effectiveness of the assay. The notified polymer was not mutagenic to bacteria under the conditions of the test. BSL (2009f)
using Bacteria. Plate incorporation procedure and Pre incubation procedure Species/Strain S. typhimurium: TA1535, TA1537, TA98 and TA100 E. coli: WP2uvrA Metabolic Activation System Phenobarbital and -naphthoflavone induced rat liver microsomal fraction (S9) in Main Test a) With metabolic activation: 31.6 – 5,000 µg/plate b) Without metabolic activation: 31.6 – 5,000 µg/plate Vehicle DMSO Remarks - Method Main Test 1 and Test 2 used plate incorporation procedure and pre incubation procedure, respectively. Positive controls: 10 µg/plate sodium azide (NaN3) for TA100 and TA1535 10 µg/plate 4-nitro-o-phenylene-diamine (4-NOPD) for TA98 40 µg/plate 4-nitro-o-phenylene-diamine (4-NOPD) for TA1537 1 µL/plate methyl methane sulfonate (MMS) for WP2uvrA 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 > 5,000 > 5,000 Negative Test 2 - > 5,000 > 5,000 Negative Present Test 1 > 5,000 > 5,000 > 5,000 Negative Test 2 - > 5,000 > 5,000 Negative Remarks - Results No precipitation and cytotoxicity of the test substance were observed up to the highest concentration tested (5,000 µg/plate) in both the absence and the presence of the metabolic activation. The positive controls produced distinct increase of revertants indicative of effectiveness of the assay. The notified polymer was not mutagenic to bacteria under the conditions of the test. BSL (2009f)