were no significant increases in other tester strains, either in the presence or absence of metabolic activation; Concurrent positive controls used in the test induced marked increases in the frequency of revertant colonies and the activity of the S9 fraction was found to be satisfactory. Additive 3 is considered mutagenic in bacterial strains Result: TA 98 and TA 1538 in the presence of metabolic activation. Investigation 2 TA98 Strain: Salmonella typhimurium Additive 3: 85% of notified chemical in ester vehicle; Test substances: Additive 3A: 50% of notified chemical in ester vehicle Acetone, THF Test Substance Vehicle: Liver fraction (S9 mix) from rats pretreated with Metabolic activation system: Aroclor 1254 Test substances were soluble in both acetone and THF at Test Substance Solubility in and above 200 mg/ml Vehicle Each concentration was tested in triplicate, with S9 as Experiment 1 – follows: Design and Test Additive 3 or Additive 3A at 0, 3.3, 10, 33, 100, 333, 1 Concentrations: 000 or 5 000 g/plate in acetone or THF; positive control: 2-aminoanthracene 1.0 g/plate; Vehicle controls: acetone, THF. Precipitation observed at 333 g per plate, but no Comment: toxicity; Additive 3 with acetone as vehicle: no significant increase in revertant colonies; Additive 3a with acetone as vehicle: non-dose responsive increase (2.2 fold at 5 000 g) in revertant colonies; Additive 3 and Additive 3a with THF as vehicle: no significant increase in revertant colonies. Each concentration was tested in triplicate, with S9 as Experiment 2 – follows: Study Design and Test Concentrations: Additive 3 at 0, 3 333 or 4 000 g/plate each in plating aliquots of 5, 10, 25, 50 100 L of acetone; positive control: 2-aminoanthracene 1.0 g/plate; vehicle control: acetone
were no significant increases in other tester strains, either in the presence or absence of metabolic activation; Concurrent positive controls used in the test induced marked increases in the frequency of revertant colonies and the activity of the S9 fraction was found to be satisfactory. Result: Additive 3 is considered mutagenic in bacterial strains TA 98 and TA 1538 in the presence of metabolic activation. Investigation 2 Strain: Salmonella typhimurium TA98 Test substances: Additive 3: 85% of notified chemical in ester vehicle; Additive 3A: 50% of notified chemical in ester vehicle Test Substance Vehicle: Acetone, THF Metabolic activation system: Liver fraction (S9 mix) from rats pretreated with Aroclor 1254 Test Substance Solubility in Vehicle Test substances were soluble in both acetone and THF at and above 200 mg/ml Experiment 1 – Design and Test Concentrations: Each concentration was tested in triplicate, with S9 as follows: Additive 3 or Additive 3A at 0, 3.3, 10, 33, 100, 333, 1 000 or 5 000 g/plate in acetone or THF; positive control: 2-aminoanthracene 1.0 g/plate; Vehicle controls: acetone, THF. Comment: Precipitation observed at 333 g per plate, but no toxicity; Additive 3 with acetone as vehicle: no significant increase in revertant colonies; Additive 3a with acetone as vehicle: non-dose responsive increase (2.2 fold at 5 000 g) in revertant colonies; Additive 3 and Additive 3a with THF as vehicle: no significant increase in revertant colonies. Experiment 2 – Study Design and Test Concentrations: Each concentration was tested in triplicate, with S9 as follows: Additive 3 at 0, 3 333 or 4 000 g/plate each in plating aliquots of 5, 10, 25, 50 100 L of acetone; positive control: 2-aminoanthracene 1.0 g/plate; vehicle control: acetone
NA705FR.docx