Ethyl acetate

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:

1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Published study but containing sufficient details to be able to judge it reliable for hazard assessment. Part of a programme of work by the NTP. Data tables available for results. TA102 not included. Full documentation in NTP studies available.

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1992

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Ethyl acetate

EC Number:

205-500-4

EC Name:

Ethyl acetate

Cas Number:

141-78-6

Molecular formula:

C4H8O2

IUPAC Name:

ethyl acetate

Details on test material:

- Name of test material (as cited in study report): Ethyl Acetate
- Analytical purity: >99%

Method

Species / strain

Species / strain 1

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

- Type and identity of media:
- Culture prepared at 37C overnight with shaking in Oxoid #2 broth or in defined minimal medium supplemented with biotin and histidine. Phenotype analysed at time of use.

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 97

Details on mammalian cell type (if applicable):

- Type and identity of media:
- Culture prepared at 37C overnight with shaking in Oxoid #2 broth or in defined minimal medium supplemented with biotin and histidine. Phenotype analysed at time of use.

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced male SD rat and male Syrian hamster liver S9 fraction, each used at two concentrations (10% and 30%)

Test concentrations with justification for top dose:

0 ; 100 ; 333 ; 1000 ; 3333 ; 10000 µg/plate. Doses were prepared using dimethyl sulphoxide as the solvent; a maximum of 0.5 ml solvent was added to each plate. Each dose was tested in triplicate without activation, and with 10% rat and hamster liver S-9.

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Migrated to IUCLID6: used for TA1535 and TA100 without metabolic activation

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

Migrated to IUCLID6: used for TA97 without metabolic activation

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylenediamine used for TA98 without metabolic activation

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene for all strains with metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 mins at 37C
- Expression time (cells in growth medium): 2 days at 37C

NUMBER OF REPLICATIONS: 5 concentrations in triplicate without metabolic activation, and with 10% and 30% liver S-9 from rat and hamster. Replicate tests were performed after the initial trial to confirm these results.

Evaluation criteria:

A material was considered mutagenic if it produced a reproducible, dose-related increase in revertants over the solvent control, under a single metabolic activation condition, in replicate trials. A material was considered questionable if the positive response was elicited at only one concentration, or if the response could not be reproduced. A chemical was designated as non-mutagenic only after it was tested without metabolic activation, and with 10% and 30% rat and hamster S-9.

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:

no cytotoxicity

Remarks:

up to maximum dose tested (10000ug/ml)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

up to maximum dose tested (10000ug/ml)

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'.

Any other information on results incl. tables

After a negative result was obtained, the substance was retested without metabolic activation and with 30% S-9. Repeat experiments were performed at least one week following the initial trial.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Executive summary:

Zeiger et al (1992) reported that ethyl acetate was negative in a Salmonella gene mutation assay (Ames Tester Strains TA97, TA98, TA100, TA1535, and TA1537), with and without exogenous metabolic activation.