Ethyl 3-[(5-chloro-2-nitrophenyl)phenylamino]-3-oxopropionate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

The test method was designed to be compatible with the OECD guidelines for Teting of Chemicals no. 471, Method B13/14 of Commission Regulation (EC) no. 440/2008 and the USA, EPA OCSPP harmonized guideline.

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Ethyl 3-[(5-chloro-2-nitrophenyl)phenylamino]-3-oxopropionate

EC Number:

244-907-1

EC Name:

Ethyl 3-[(5-chloro-2-nitrophenyl)phenylamino]-3-oxopropionate

Cas Number:

22316-45-6

Molecular formula:

C17H15ClN2O5

IUPAC Name:

ethyl 3-[(5-chloro-2-nitrophenyl)(phenyl)amino]-3-oxopropanoate

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:

Exp 1: 1.5 to 5000 ug/plate
The dose range was amended following the results of Exp.1 and was 15 to 5000 ug/plate.

Vehicle / solvent:

dimethyl sulphoxide

Details on test system and experimental conditions:

Six dose levels were selected in Exp 2 in order to achieve both a minimum of four non-toxic dose levels and the toxic of the test item following the change in test methodology.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium, other: TA1535, TA1537, TA98 and TA100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Any other information on results incl. tables

No visible reduction in the growth of the bacterial background lawn at any dose level, either in the presence or absence of metabolic activation, in the first mutation test (plate incorporation method) and consequently the same maximum dose level was used in the second mutation test. Similarily, there was no visible reduction in the growth of the bacterial background lawn at any dose level, either in the presence or absence of metabolic activation, in the second mutation test (pre-incubation method). A test item film was noted at 5000 ug/plate in the first mutation test only (plate incorporation method9 this observation did not prevent the scoring of revertant colonies.

There were no significant increases in the frequency of revertant colonies recorded for any of the bacterial strains, with any dose of the test item, either with of without S9-mix in Exp.1 (plate incorporation method). Similarly, no toxicologically significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test item either with or witohout S9mix in Exp. 2 (pre-incubation method).

A small, statistically significant increase in revertant colony frequency was observed in the second mutation test in TA1535 in the absence of S9 -mix at 15 ug/plate. This increase was considered to be of no biological relevance because there was no evidence of a dose-response relationship or reproducibility. Furthermore, the individual revertant counts at the statistically significant dose level within the in-house historical untreated/vehicle control rang for each tester strain and the maximum fold increase was only 1.8 times the concurrent vehicle controls.

Applicant's summary and conclusion

Conclusions:

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