2'-methylacetoacetanilide

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study documented and performed according to GLP standards and in compliance with OECD Test Guidelines 471 and 472.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

not specified

Principles of method if other than guideline:

Not applicable.

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2'-methylacetoacetanilide

EC Number:

202-267-0

EC Name:

2'-methylacetoacetanilide

Cas Number:

93-68-5

Molecular formula:

C11H13NO2

IUPAC Name:

N-(2-methylphenyl)-3-oxobutanamide

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Purity: 99.9%

Method

Target gene:

Not applicable.

Species / strain

Species / strain / cell type:

other: TA98, TA100, TA1535, TA1537, Escherichia coli WP2uvrA

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9 from male rat liver, induced with phenobarbital and 5,6-benzoflavon

Test concentrations with justification for top dose:

0, 156, 313, 625, 1250, 2500 and 5000 microg/plate.

Vehicle / solvent:

Dimethylsulphoxide

Controls

Untreated negative controls:

yes

Remarks:

dimethylsulphoxide

Negative solvent / vehicle controls:

yes

Remarks:

dimethylsulphoxide

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Without S9mix: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (TA 98, TA 100 and WP2), sodium azide (TA 1535), 9-aminoacridine hydrochloride (TA 1537); With S9 mix: 2-aminoanthracene (all five strains)

Details on test system and experimental conditions:

TEST SYSTEM:
Metabolic activation system: S9 from male rat liver, induced with phenobarbital and 5,6-benzoflavon
ADMINISTRATION:
Number of replicate: 2
Plates per dose: 3
Application: Pre-incubation
Solvent: DMSO (Concentration was not described)
Test parameter: Revertant colonies per plate

Evaluation criteria:

Number of revertant colonies for all treatment groups is compared with those obtained for solvent control groups.
Any toxic effects of the test substance can be detected by a substantial reduction in revertant colony counts or by
the absence of a complete background bacterial lawn.

Statistics:

Not applicable.

Results and discussion

Test results

Species / strain:

other: TA98, TA100, TA1535, TA1537, Escherichia coli WP2uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Number of revertant colonies per plate in all doses with/without S9mix were equivalent to control. On the other hand, more than two times revertant colonies were observed in all positive controls. Visible precipitation was not observed in any plates.

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 without metabolic activation
negative with metabolic activation

lt is concluded that, when tested at dose levels up to 5000 microg/plate in dimethylsulphoxide, Acetoacet-o-toluidide was not mutagenic in this bacterial test system.

Executive summary:

The genetic toxicity study (Ames) was published in the OECD SIDS dossier in year 2003 conducted and reported

according to the OECD Guideline for the testing of chemicals 471 and 472 and principles of GLP in year 1999. Number of revertant colonies per plate in all doses with/without S9 mix were equivalent to control. On the other hand, more than two times revertant colonies were observed in all positive controls. Visible precipitation was not observed in any plates.

When tested at dose levels of up to 5000 microg/plate in five strains with or without S9 mix no toxicity was observed. A top dose level of 5000 g/plate was chosen for the subsequent mutation study. Other dose levels used in the mutation assays were: 2500, 1250, 625, 313, 156 microg/plate. The concurrent positive control compounds demonstrated the sensitivity of the assay and the metabolising activity of the liver preparations.

Therefore, Acetoacet-o-toluidide is not mutagenic to Salmonella typhimurium and Escherichia coli.