1,2,4-Benzenetricarboxylic acid, mixed branched tridecyl and isodecyl esters

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Conducted according OECD TG 471. GLP.

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 3

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,2,4-Benzenetricarboxylic acid, tris(2-ethylhexyl) ester

IUPAC Name:

1,2,4-Benzenetricarboxylic acid, tris(2-ethylhexyl) ester

Constituent 2

Reference substance name:

Tris(2-ethylhexyl) benzene-1,2,4-tricarboxylate

EC Number:

222-020-0

EC Name:

Tris(2-ethylhexyl) benzene-1,2,4-tricarboxylate

Cas Number:

3319-31-1

Molecular formula:

C33H54O6

IUPAC Name:

tris(2-ethylhexyl) benzene-1,2,4-tricarboxylate

Details on test material:

CAS #3319-31-1; 1,2,4-Benzenetricarboxylic acid, tris(2-ethylhexyl) ester

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

Test concentrations with justification for top dose:

without S9: 0, 313, 625, 1250, 2500, 5000 ug/plate (five strains)
with S9: 0, 313, 625, 1250, 2500, 5000 ug/plate (five strains)

Vehicle / solvent:

Acetone

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

Acetone, DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Without S9 mix, 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (TA100, WP2, TA98) Sodium azide (TA1535) 9-Aminoacridine (TA 1537); With S9 mix, 2- Aminoanthracene (five strains)

Details on test system and experimental conditions:

Prior to assay initiation, a toxicity pretest was performed using tester strain TA100. Based on these results, the doses for the final assay were determined. Each chemical was tested initially at half-log dose intervals up to a dose that elicited toxicity, or to a dose immediately below one which was toxic in the preliminary toxicity test. Chemicals that were not toxic were tested, with few exceptions, to a maximum dose of 10 mg/plate. In the definitive assay, each of the five strains was dosed with either the test substance; a vehicle control (DMSO); or a nontreated control and a positive control. The test mixture containing the tester strain and test substance with or without S9 was added to the surface of petri dishes containing Vogel-Bonner medium. The S-9 (9,000g supernatant) fractions of Aroclor 1254-induced, male Sprague-Dawley rat and male Syrian hamster livers were prepared as described previously [Haworth et al, 1983]. The S-9 mixes were prepared immediately prior to use and contained either 10% or 30% S-9; occasionally, other levels were used. The histidine-independent colonies that formed on the plates were counted following a two-day incubation at 37°C. Positive controls were as follows: 2 aminoanthracene (all strains with S9); sodium azide (without S9, TA1535, TA100), 4-nitro-ophenylenediamine (without S9, TA98) and 9-aminoacridine (without S9, TA 97, TA1537). There were 3 plates/dose group/strain/treatment. The test results were verified by repeating the assay. If the results were negative, they were repeated first without S9 and then with 30% S9.

Species/Strain: Salmonella typhimurium TA100, TA1535, TA98, TA1537; Escherichia coli WP2 uvrA

Positive controls:
Without S9 mix, 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (TA100, WP2, TA98) Sodium azide (TA1535)
9-Aminoacridine (TA 1537)
With S9 mix, 20 Aminoanthracene (five strains)

S9: Rat liver, induced with phenobarbital and 5,6-benzoflavone

Evaluation criteria:

Chemicals were judged to be mutagenic if the test results produced a dose-related, reproducible increase in histidine revertants over control. It was not a requirement for mutagenic responses to reach two-fold over background.

Results and discussion

Test results

Test results 1

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

no genotoxic effects were observed at the highest dose tested (5000 ug/plate)

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

Toxicity was not observed up to 5000 ug/plate in five strains with and without metabolic activation (S9 mix).

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

no genotoxic effects were observed at the highest dose tested (5000 ug/plate)

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

Toxicity was not observed up to 5000 ug/plate in five strains with and without metabolic activation (S9 mix).

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with metabolic activation at the highest dose tested (5000 ug/plate)
negative without metabolic activation at the highest dose tested (5000 ug/plate)

No genotoxic effects were observed at the highest dose tested (5000 ug/plate) in all five strains of bacteria, with or without metabolic activation.

Classification as a genotoxic agent is not warranted under the new Regulation (EC) 1272/2008 on classification, labeling and packaging of substances and mixtures (CLP) or under Directive 67/548/EEC for dangerous substances and Directive 1999/45/EC for preparations.

Executive summary:

The test material did not induce a statistically signifiant increase in revertant numbers in any of the test strains, either in the absence or in the presence of S-9. This study was therefore considered to have provided no indication of any test material mutagenic activity.

Classification as a genotoxic agent is not warranted under the new Regulation (EC) 1272/2008 on classification, labeling and packaging of substances and mixtures (CLP) or under Directive 67/548/EEC for dangerous substances and Directive 1999/45/EC for preparations.