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EC number: 240-357-1 | CAS number: 16245-77-5
The test substance was assayed for mutation in five histidine-requiring strains (TA98, TA100, TA1535, TA1537 and TA102) of Salmonella typhimurium, both in the absence and in the presence of metabolic activation by an Aroclor 1254-induced rat liver postmitochondrial fraction (S-9), in two separate experiments.
An initial toxicity range-finder experiment was carried out in the absence and in the presence of S-9 in strain TA100 only, using final concentrations of the test substance at 1.6, 8, 40, 200, 1000 and 5000 μg/plate, plus negative (solvent) and positive controls. Following these treatments, no evidence of toxicity was observed. Experiment 1 treatments of the remaining test strains were performed in the absence and in the presence of S-9 and retained the same test doses as employed for the range-finder experiment. Following these treatments, evidence of toxicity was observed, but was limited to 5000 µg/plate treatments in strain TA102 in the absence and in the presence of S-9. Experiment 2 treatments of all the tester strains were performed in the absence and in the presence of S-9 with the maximum test dose of 5000 μg/plate retained. A narrowed dose range was employed (156.3 - 5000 µg/plate), in order to examine more closely those concentrations of the test substance approaching the maximum test dose and therefore, considered most likely to provide evidence of any mutagenic activity. In addition, all treatments in the presence of S-9 were further modified by the inclusion of a preincubation step. In this way, it was hoped to increase the range of mutagenic chemicals that could be detected using this assay system. Following an increase in revertants observed in strain TA98 in the presence of S-9 in Experiment 1, plate incorporation treatments of strain TA98 in the presence of S-9 were also performed in Experiment 2. Following the Experiment 2 treatments, evidence of toxicity was again observed in strain TA102, occurring at 2500 and 5000 µg/plate in this strain in the presence of S-9, but only at 5000 µg/plate in the absence of S-9. Similar evidence of toxicity was also observed following plate incorporation treatments of strain TA98 in the presence of S-9 at the maximum test dose of 5000 μg/plate.
The test article was completely soluble in the aqueous assay system at all concentrations treated, in each of the experiments performed. Negative (solvent) and positive control treatments were included for all strains in both experiments. The mean numbers of revertant colonies on negative control plates all fell within acceptable ranges, and were significantly elevated by positive control treatments. Following the test substance treatments of all the tester strains, in the absence and in the presence of S-9, increases in revertant numbers were observed in strain TA98 in the presence of S-9 which were statistically significant when the data were analysed at the 1% level using Dunnett's test. These increases were dose-related and reproducible over two independent experiments, and were therefore considered to be indicative of the test substance’s mutagenic activity in this strain following metabolic activation. No comparable increases were seen in strain TA98 in the absence of S-9, and no other increases sufficient to be considered as indicative of mutagenic activity were observed in any of the other tester strains.
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