Ethyl 4,4,4-trifluoroacetoacetate

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:

August - September 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-compliant guideline study

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

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:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Ethyl 4,4,4-trifluoroacetoacetate

EC Number:

206-750-7

EC Name:

Ethyl 4,4,4-trifluoroacetoacetate

Cas Number:

372-31-6

Molecular formula:

C6H7F3O3

IUPAC Name:

ethyl 4,4,4-trifluoro-3-oxobutanoate

Test material form:

other: clear liquid

Details on test material:

- Name of test material (as cited in study report): P5117
- Physical state: clear, colourless liquid
- Purity test date: 25-Jul-1995
- Expiration date of the lot/batch: no data
- Stability under test conditions: stable
- Storage condition of test material: stored at ambient temperature

Method

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA98, TA1538, TA100, TA1535 and TA1537

Details on mammalian cell type (if applicable):

Type and identity of media:
Cultures of the histidine-dependent strains of Salmonella typhimurium were derived from cultures provided by Prof. Bruce Ames, University of California

Additional strain / cell type characteristics:

DNA polymerase A deficient

Metabolic activation:

with and without

Metabolic activation system:

S9 liver fractions of rats treated with Aroclor 1254

Test concentrations with justification for top dose:

- Pretest: 5, 50, 500, 2500, 5000 ug/plate
- Main test: 50, 158, 500, 1580, 5000 ug/plate

Vehicle / solvent:

Dimethylsulfoxide

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

other: 2-aminoanthracene

Details on test system and experimental conditions:

A solution of P5117 was prepared in DMSO at 50 mg/ml, and four half-log dilutions were prepared from this solution. Aliquots (0.1 ml) of each concentration of P5117 were placed in sterile tubes. Molten, histidinedeficient top-agar (2 ml), maintained at 45°C, and bacterial suspension (0.1 ml) were then added. The tubes were mixed by inversion and 0.5 ml rat liver microsomal preparation (S9 mix) or 0.1M sodium phosphate buffer was added where appropriate. The tubes were again inverted to mix thoroughly and the contents poured onto plates containing solidified minimal medium (20 ml). Further plates were prepared without the inclusion of the test organisms to verify the sterility of the S9 mix and the test material. A control series of plates was prepared to confirm the inability of DMSO (0.1 ml) to induce reversion in the bacterial strains, and to provide a measure of the spontaneous mutation rates. Aliquots (0.1 ml) of a 10 Exp.-06 dilution of the overnight cultures were spread on the surface of plates of complete medium to measure the viability and celldensity of each culture.
All plates were prepared in triplicate, allowed to solidify and incubated at 37°C for 2 days. After incubation, numbers of revertant colonies were counted, either manually or with an automated colony counter. Total colonies on nutrient plates were counted in the same way. Growth of the background lawn of non-revertant cells on minimal plates was verified. Results obtained with all strains were confirmed in a second, independent experiment. All plates and tubes were identified by the use of numbers indelibly marked
on the plates and test tube racks.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA98, TA1538, TA100, TA1535 and TA1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

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
negative without metabolic activation

It was concluded that P5117 did not exhibit any mutagenic activity under the conditions of test.

Executive summary:

This reverse mutation assay was performed in 1995 as GLP-study in accordance with OECD- and EU-guidelines. Five Salm. typh. strains (TA1535, TA1537, TA1538, TA98 and TA100) were tested with and without metabolic activation. Test concentrations ranged from 50 ug/plate up to 5000 ug/plate. The test item was found to be not mutagenic in this bacterial reversion mutation assay.