WP2uvrA- E. coli: Metabolic Activation System S9 mix from induced rat liver phenobarbitone/β-naphthoflavone in a) With metabolic activation: 5 5,000 µg/plate – Main Test b) Without metabolic activation: 5 5,000 µg/plate – Vehicle Distilled water Remarks - Method A preliminary test and range-finding test (test 1) were performed to determine the concentration of test substance in the main test (Test 2). Positive controls: with metabolic activation - 2-Aminoanthracene [TA100, TA1535, TA1537, WP2uvrA-], Benzo(a)pyrene [TA98]; without metabolic activation N-ethyl-N’-nitro-N-nitrosoguanidine [WP2uvrA-, – TA100, TA1535], 9-Aminoacridine [TA1537], 4-Nitroquinoline-1-oxide [TA98]. Metabolic Test Substance Concentration (µg/plate) Resulting in: Activation Cytotoxicity in Cytotoxicity in Precipitation Genotoxic Effect Preliminary Test Main Test Absent Test 1 500 500 1,500 negative ≥ ≥ ≥ Test 2 500 1,500 negative ≥ ≥ Present Test 1 500 5,000 1,500 negative ≥ ≥ ≥ Test 2 5,000 1,500 negative ≥ ≥ Remarks - Results No biologically relevant increase in the frequency of revertant colonies was obtained in the presence or absence of metabolic activation. Growth inhibition and a significant reduction in the number of revertants was observed in all strains in the presence and absence of S. typhimurium metabolic activation from 500 µg/plate. A decrease in the number of revertant colonies was also observed for in the presence and E. coli absence of metabolic activation. However, this decrease did not always relate to a corresponding increase in test substance. The precipitate observed at and above 1,500 µg/plate did not prevent scoring of revertant colonies. Positive and negative controls performed as expected. The notified chemical was not mutagenic to bacteria under the conditions of the test. Safepharm (2008e)
E. coli: WP2uvrA- Metabolic Activation System S9 mix from phenobarbitone/β-naphthoflavone induced rat liver in Main Test a) With metabolic activation: 5 – 5,000 µg/plate b) Without metabolic activation: 5 – 5,000 µg/plate Vehicle Distilled water Remarks - Method A preliminary test and range-finding test (test 1) were performed to determine the concentration of test substance in the main test (Test 2). Positive controls: with metabolic activation - 2-Aminoanthracene [TA100, TA1535, TA1537, WP2uvrA-], Benzo(a)pyrene [TA98]; without metabolic activation – N-ethyl-N’-nitro-N-nitrosoguanidine [WP2uvrA-, TA100, TA1535], 9-Aminoacridine [TA1537], 4-Nitroquinoline-1-oxide [TA98]. Metabolic Activation Test Substance Concentration (µg/plate) Resulting in: Cytotoxicity in Preliminary Test Cytotoxicity in Main Test Precipitation Genotoxic Effect Absent Test 1 ≥ 500 ≥ 500 ≥ 1,500 negative Test 2 ≥ 500 ≥ 1,500 negative Present Test 1 ≥ 500 ≥ 5,000 ≥ 1,500 negative Test 2 ≥ 5,000 ≥ 1,500 negative Remarks - Results No biologically relevant increase in the frequency of revertant colonies was obtained in the presence or absence of metabolic activation. Growth inhibition and a significant reduction in the number of revertants was observed in all S. typhimurium strains in the presence and absence of metabolic activation from 500 µg/plate. A decrease in the number of revertant colonies was also observed for E. coli in the presence and absence of metabolic activation. However, this decrease did not always relate to a corresponding increase in test substance. The precipitate observed at and above 1,500 µg/plate did not prevent scoring of revertant colonies. Positive and negative controls performed as expected. The notified chemical was not mutagenic to bacteria under the conditions of the test. Safepharm (2008e)