Ethylene diacetoacetate

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: Performed according to a previous guideline version using a different combination of test strains

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Ethylene diacetoacetate

EC Number:

226-724-9

EC Name:

Ethylene diacetoacetate

Cas Number:

5459-04-1

Molecular formula:

C10H14O6

IUPAC Name:

ethane-1,2-diyl bis(3-oxobutanoate)

Details on test material:

- Name of test material (as cited in study report): Acetessigsäureglykolester

Method

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver homogenate fraction (S9)

Test concentrations with justification for top dose:

4, 20, 100, 500, 2500, 5000 µg/plate

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Sodium-azide, 9-Aminoacridine, 2-Nitrofluorene, 2-Aminoanthracene

Evaluation criteria:

A test compound is classified as mutagenic if it has either of the following effects:

a) it produces at least a 2-fold increase in the mean number of revertants per plate of
at least one of the tester strains over the mean number of revertants per plate of
the appropriate vehicle control at complete bacterial background lawns.

b) it induces a dose-related increase in the mean number of revertants per plate of at
least one of the tester strains over the mean number of revertants per plate of the
appropriate vehicle control in at least two to three concentrations of the test
compound at complete bacterial background lawns.

If the test substance does not produce reproducible increases of at least 2 times the
concurrent solvent controls, at any dose level with any bacterial strain, it is considered
to show no evidence of mutagenic activity in this system.

The test results must be reproducible.

Results and discussion

Test results

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:

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

Acetessigsaureglycolester is not mutagenic in these bacterial test systems either in the absence or in the presence of an exogenous metabolizing system.

Executive summary:

Acetessigsäureglycolester was tested for mutagenicity with the strains TA 100, TA 1535, TA 1537 and TA 98 of Salmonella typhimurium.

Two independent mutagenicity studies were conducted, each in the absence and in the presence of a metabolizing system derived from a rat liver homogenate. For both studies, the compound was dissolved in DMSO, and each bacterial strain was exposed to 6 dose levels. Doses for both studies ranged from 4 to 5000 µg/plate.

Control plates without mutagen showed that the number of spontaneous revertant colonies was within the laboratory's historical control range and similar to that described in the literature. All the positive control compounds gave the expected increase in the number of revertant colonies.

Toxicity: In the mutagenicity experiments toxicity was not observed either with or without metabolic activation. In the toxicity test using histidine enriched agar plates and a dilution of the tester strain TA 100 (designated TA 100 D), which was performed in parallel with the second experiment, toxicity was found at concentrations of 2500 µg/plate and above in the absence of metabolic activation. In the presence of metabolic activation the test compound proved to be not toxic to the bacterial strain.

Mutagenicity: In the absence and in the presence of the metabolic activation system Acetessigsaureglycolester did not result in relevant increases in the number of revertants in any of the bacterial strains.

Summarizing, it can be stated that Acetessigsäureglycolester is not mutagenic in these bacterial test systems either with or without exogenous metabolic activation at the dose levels investigated.