Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed publication

Data source

Reference
Reference Type:
publication
Title:
Gene mutation toxicity study for the test chemical
Author:
Ishidate et al
Year:
1984
Bibliographic source:
Food and chemical toxicology

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
A salmonella/microsome test (Ames tests) was performed on the test substance to evaluate its mutagenic effect.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: Ethyl butyrate
- IUPAC name: Ethyl butyrate
- Molecular formula: C6H12O2
- Molecular weight: 116.16 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: 99.8%
- Impurities (identity and concentrations): 0.2%

Method

Target gene:
Histidine
Species / strain
Species / strain / cell type:
other: TA92, TA1535,TA100, TA1537, TA94 and TA98
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data available
Metabolic activation:
with and without
Metabolic activation system:
The liver microsome fraction (S-9) was prepared from the liver of Fischer rats
Test concentrations with justification for top dose:
Six different concentrations were used with 10 mg/plate as the maximum concentration.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The chemical was soluble in DMSO
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
not specified
Positive control substance:
not specified
Details on test system and experimental conditions:
METHOD OF APPLICATION: Preincubation

DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: Duplicate

NUMBER OF CELLS EVALUATED: No data available

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available

OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available

OTHER: No data available
Rationale for test conditions:
No data available
Evaluation criteria:
The result was considered positive if the number of colonies found was twice the number in the control (exposed to the appropriate solvent or untreated).
Statistics:
No data available

Results and discussion

Test results
Species / strain:
other: TA92, TA1535,TA100, TA1537, TA94 and TA98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
The maximum dose for negative results represents the highest non-cytotoxic dose used in the experiment.
Remarks on result:
other: No mutagenic potential

Applicant's summary and conclusion

Conclusions:
The test chemical did not induce a doubling of His + revertant colonies over the control using S. typhimurium TA92, TA1535, TA100, TA1537, TA94, TA98 strains in the presence and absence of metabolic activation system. Hence, Ethyl butyrate is considered as Non-mutagenic (negative) in vitro in the test system used under the certain experimental conditions.
Executive summary:

A bacterial reverse mutation test was performed to determine the mutagenic nature of the test substance. The study was performed using Salmonella typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 with and without S9 metabolic activation system. The test was performed as a preincubation assay at six different concentration with 10 mg/plate being the maximum concentration.Cells were pre-incubated with both the test chemical and S9-mix for 20 min. The number of revertant (His+) colonies was scored after 48 hours of incubation. The result was considered positive if the number of revertant colonies was found twice the number of the controls(exposed to the appropriate solvent or untreated). The test chemical did not induce a doubling of revertant colonies over the control using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 in the presence and absence of S9 metabolic activation system. Hence, Ethyl butyrate is considered as Non-mutagenic (negative)in vitro in the test system used under the certain experimental conditions.