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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
No data
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference
Reference Type:
publication
Title:
Salmonella Mutagenicity Tests: II. Results from the Testing of 270 Chemicals.
Author:
Mortelmans K., Haworth S., Lawlor T., Speck W., Tainer B., Zeiger E.
Year:
1986
Bibliographic source:
Environmental Mutagenesis Volume 8, Supplement 7: 1-119
Report Date:
1985

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
Benzyl Acetate was examined alongside 269 other chemicals for its ability to induce mutagenic changes when tested in Salmonella typhimurium bacterial strains in the presence and absence of metabolic activation with rat and hamster S-9 mix using the preincubation assay method.
GLP compliance:
not specified
Remarks:
Study is a publication
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Benzyl Acetate
- Molecular formula (if other than submission substance): No information provided
- Molecular weight (if other than submission substance): No information provided
- Smiles notation (if other than submission substance): No information provided
- InChl (if other than submission substance): No information provided
- Structural formula attached as image file (if other than submission substance): see Fig. No information provided
- Substance type: No information provided
- Physical state: No information provided
- Analytical purity: 99.1%
- Impurities (identity and concentrations): No information provided
- Composition of test material, percentage of components: No information provided
- Isomers composition: No information provided
- Purity test date: No information provided
- Lot/batch No.: No information provided
- Expiration date of the lot/batch: No information provided
- Radiochemical purity (if radiolabelling): No information provided
- Specific activity (if radiolabelling): No information provided
- Locations of the label (if radiolabelling): No information provided
- Expiration date of radiochemical substance (if radiolabelling): No information provided
- Stability under test conditions: No information provided
- Storage condition of test material: No information provided
- Other: No information provided
Specific details on test material used for the study:
- Name of test material (as cited in study report): Benzyl Acetate
- Substance type: No information provided
- Physical state: clear liquid
- Analytical purity: 99.1%

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
Not applicable as the species/strain used are bacterial and not mammalian.
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium TA 97
Details on mammalian cell type (if applicable):
Not applicable as the species/strain used are bacterial and not mammalian.
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced rat and hamster metabolic activation (S-9 mix).
Test concentrations with justification for top dose:
0,10,100,1000 and 10000ug/plate.
Vehicle / solvent:
Distilled water. For chemicals that were not soluble or had low solubility in water, dimethyl sulfoxide (DMSO) was used. Ethanol (95%) or acetone was used for chemicals insoluble in water or DMSO.
Controlsopen allclose all
Untreated negative controls:
yes
Remarks:
Potassium chloride
Negative solvent / vehicle controls:
not specified
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
Used with metabolic activation for strains TA 1535 and TA 100
Untreated negative controls:
yes
Remarks:
Potassium chloride
Negative solvent / vehicle controls:
not specified
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylenediamine
Remarks:
Used with metabolic activation for strain TA 98
Untreated negative controls:
yes
Remarks:
Potassium chloride
Negative solvent / vehicle controls:
not specified
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
Used with metabolic activation for strains TA 97 and TA 1537
Untreated negative controls:
yes
Remarks:
Potassium chloride
Negative solvent / vehicle controls:
not specified
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
Used with all strains with rat and hamster liver metabolic activation systems.
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48 hours
- Expression time (cells in growth medium): Not documented
- Selection time (if incubation with a selection agent): Not documented
- Fixation time (start of exposure up to fixation or harvest of cells): Not documented

SELECTION AGENT (mutation assays): Not documented
SPINDLE INHIBITOR (cytogenetic assays): Not documented
STAIN (for cytogenetic assays): Not documented

NUMBER OF REPLICATIONS: All assays were repeated no less than one week after completion of the initial test.

NUMBER OF CELLS EVALUATED: Not documented

DETERMINATION OF CYTOTOXICITY
- Method: Toxicity was evidenced by one or more of the following phenomena: appearance of his- pinpoint colonies, reduced numbers of revertant colonies per plate or thinning or absence of the bacterial lawn.

OTHER EXAMINATIONS:
- Determination of polyploidy: Not documented
- Determination of endoreplication: Not documented
- Other: Not documented

OTHER: At least one toxic dose was incorporated into the first mutagenicity test, the repeat test(s) occasionally had the doses adjusted so that an apparent toxic dose was not reached.
Evaluation criteria:
Mutagenic response: a dose-related, reproducible increase in the number of revertants over background, even if the increase was less than twofold.
Nonmutagenic response: when no increase in the number of revertants was elicited
Questionable response: when there was an absence of a clear cut dose related increase in revertants, when the dose related increase in revertants was not reproducible or when the response was of insufficient magnitude to support a determination of mutagenicity.
Statistics:
No information provided.

Results and discussion

Test resultsopen allclose all
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:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Species / strain:
S. typhimurium TA 97
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
The hamster S-9 mix proved, in general, to be better than the rat S-9 mix in inducing a mutagenic response
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

No additional information provided.

Applicant's summary and conclusion

Conclusions:
Based on the results of the study, the test substance, Benzyl Acetate can be considered to be non-mutagenic under the conditions of this study. As a result of this, it does not require any classification according to Regulation EC No. 1272/2008.
Executive summary:

In the study conducted by Mortelmans et al in 1986, Benzyl Acetate was examined for its ability to cause mutagenic changes when tested in five strains of the bacteria Salmonella typhimurium, specifically, TA 1535, TA 1537, TA97, TA 98 and TA 100 through the preincubation assay method. The test was conducted both in the presence and absence of metabolic activation using rat and hamster liver derived S-9 mix, over a range of doses, from 0 to 10000 ug/plate. Based on the results of this study, the test substance Benzyl Acetate was not considered to be mutagenic under the conditions of this test. As a result of this, it does not require any classification according to Regulation EC No. 1272/2008.