Reaction mass of benzenamine, 2-ethyl-N-(2-ethyl-nonylphenyl) nonyl-, branched and benzenamine, 2-ethyl-N-(2-ethyl-nonylphenyl)-, branched

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 December 1996 to 16 May 1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of benzenamine, 2-ethyl-N-(2-ethyl-nonylphenyl) nonyl-, branched and benzenamine, 2-ethyl-N-(2-ethyl-nonylphenyl)-, branched

Molecular formula:

Not applicable

IUPAC Name:

Reaction mass of benzenamine, 2-ethyl-N-(2-ethyl-nonylphenyl) nonyl-, branched and benzenamine, 2-ethyl-N-(2-ethyl-nonylphenyl)-, branched

Test material form:

liquid

Details on test material:

CAS number: Not yet known
EC Number: Not yet known

Specific details on test material used for the study:

Description: Dark amber viscous liquid
Purity: 100%

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9 Rat liver, induced with Aroclor

Test concentrations with justification for top dose:

Test 1 and Test 2: 0, 50, 150, 500, 1500 and 500 µg/plate.
5000 µg is the standard top dose recommended in the guidelines.

Vehicle / solvent:

Acetone

Controls

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

N-ethyl-N-nitro-N-nitrosoguanidine

other: 2-Aminoanthracene

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium, other: TA 100, A 1535, TA 1537 and TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

1500 and 5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

1500 and 5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

The test material was considered to be non-mutagenic under the test conditions of this test.

Executive summary:

Salmonella typhimurium strains TA 1535, TA 1537, TA98 and TA 100, and Escherichia coli strain WP2uvrA- were treated with the test material using the Ames plate incorporation method at five dose levels, 50, 150, 500, 1500, and 5000 ug/plate, in triplicate, both with and without the addition of a rat liver S9 metabolising system. The experiment was repeated on a separate day using the same dose range as experiment 1, fresh cultures of the bacterial strains, and fresh test material formulations.

The dose range was determined in a preliminary toxicity assay in which the test material at dose levels from 50 to 5000 µg/plate caused no visible reduction in the growth of the bacterial lawn at any dose level, either with or without metabolic activation.  A precipitate was observed at and above 1500 µg/plate, but did not interfere with the scoring of revertant colonies.

No significant increase in the frequency of revertant colonies was recorded for any of the bacterial strains with any dose of the test material, either with or without metabolic activation. The vehicle (acetone) control plates produced counts of revertant colonies within the normal range.  All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with and without the metabolising system. The test material was considered to be non-mutagenic under the conditions of this test.