4'-chloro-2',5'-dimethoxyacetoacetanilide

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Important aspects in line with current OECD guidelines. Prior to GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1982

Report date:

1982

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Company guideline similar to OECD 471

Deviations:

not applicable

Principles of method if other than guideline:

This report describes experiments performed in a short term test using the procedure of the Salmonella / mammalian-microsome-mutagenicity test (Ames Test) to assess the mutagenic potential of the test material in amino acid-dependent strains of Salmonella typhimurium and a strain of Escheriachia coli described by Green.

References:
1) B.N. Ames, W.W. Durston, E. Yamasaki and F.D. Lee, Carcinogens are mutagens. A simple test system combining liver homogenate for activation and bacteria for detection, Proc. Nat. Acad. Sci., USA 70 (1973) 2281-2285

2) B.N. Ames, J. McCann and E. Yamasaki, Methods for detecting carcinogens and mutagens with the Salmonella / mammalian-microsome mutagenicty test, Mutat. Res. 31 (1975) 347-364

3) M.H.L. Green and W.J. Muriel, Mutagen testing using trp+ reversion in Escherichia coli, Res. 38 (1976) 3-32

4) A. P. Alvares, D.R. Bickers and A. Kappas: Poly chlorinated biphenyls: a new type of inducer of cytochrome P 448 in the liver. Proc. Nat. Acad. Sci USA 70 (1973) 1321-1325

GLP compliance:

no

Remarks:

- prior to EU GLP Directives

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4'-chloro-2',5'-dimethoxyacetoacetanilide

EC Number:

224-638-6

EC Name:

4'-chloro-2',5'-dimethoxyacetoacetanilide

Cas Number:

4433-79-8

Molecular formula:

C12H14ClNO4

IUPAC Name:

N-(4-chloro-2,5-dimethoxyphenyl)-3-oxobutanamide

Details on test material:

- Name of test material (as cited in study report): Naphtol AS-IRG Pt. 288/82

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:

S. typhimurium TA 1538

Species / strain 3

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:

4 to 10000 µg/plate (toxicity test)
4 to 5000 µg/plate (mutagenicity)

Vehicle / solvent:

DMSO

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA 100, TA 1535: without metabolic activation

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA 1537: without metabolic activation

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA 98, TA 1538: without metabolic activation

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-Methyl-N'-nitro-N-nitrosoguanidine

Remarks:

WP2uvrA: without metabolic activation

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

TA 98, TA100, TA1535, TA 1537, TA 1538, WP2uvrA: with metabolic activation

Controls 6

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene

Remarks:

TA 98, TA100, TA1535, TA 1537, TA 1538, WP2uvrA: with metabolic activation

Details on test system and experimental conditions:

- Agar mixed with histidine (Salmonella) / tryptophan (E.coli) and biotin solution.
- The test compound solution, overnight nutrient broth culture and if required S-9 mix or buffer added.
- After incubation for 48 to 72 hrs at 37°C in the dark, colonies (his+ revertants) are counted.
- Preliminary toxicity tests were performed with all tester strains (reduced rate of spontaneously occurring colonies as well as visible thinning of the bacterial lawn were used as indicator for toxicity)

Evaluation criteria:

positive: significant increase in the number of colonies

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

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

At doses between 5000µg/plate and 10000µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

At doses between 5000µg/plate and 10000µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

At doses between 5000µg/plate and 10000µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

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

Summarizing, it can be stated that Naphtol AS IRG Pt. 288/82 is not mutagenic in these bacterial test systems either with or without exogenous metabolic activation at the dose levels investigated.

Executive summary:

Naphtol AS IRG Pt. 288/82 was tested for mutagenicity with the strains TA 100, TA 1535, TA 1537, TA 1538, TA 98 of Salmonella typhimurium and Escherichia coli WP2uvrA.

The mutagenicity studies were conducted in the absence and in the presence of a metabolizing system derived from rat liver homogenate. A dose range of 6 different doses from 4 µg/plate to 5 000 µg/plate was used.

Control plates without mutagen showed that the number of spontaneous revertant colonies was similiar to that described in the literature. All the positive control compounds gave the expected increase in the number of revertant colonies.

Toxicity: The test compound proved to be toxic to the bacteria at 5 000 or 10 000 µg/ plate. 5 000 µg/plate was chosen as top dose level for the mutagenicity study.

Mutagenicity: In the absence of metabolic activation system the test compound did not show a significant influence on the number of revertants in any of the bacterial strains due to mutagenicity. Also in the presence of a metabolic activation system, treatment of the cells with Naphtol AS-IRG Pt. 288/82 did not result in an increase in the number of revertant colonies with any of the strains used.

Summarizing, it can be stated that Naphtol AS IRG Pt. 288/82 is not mutagenic in these bacterial test systems either with or without exogenous metabolic activation at the dose levels investigated.