Ethyl formate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Data for test chemicals was reviewed to determine the mutagenic nature of ethyl formate (109-94-4). The studies are as mentioned below:

In Vitro Genetic Mutation study

AMES Assay

Mutagenicity potential of test substance was determined in bacterial strain by AMES assay. The test substance was exposed to salmonella strains TA98 and TA 100 with and without metabolic activation at concentrations of 0, 1000, 2500, 5000, 7500, 10000, µg/plate. No mutagenic effects were observed in the study both in the presence and absence of metabolic activation. Based on the results obtained the test substance was considered to be non-mutagenic with and without activation. Hence the substance cannot be classified as gene mutant in vitro.

In vitro Chromosomal abbreviation study in mammalian cell

The test chemical did not induce chromosomal aberration in the Chinese hamster fibroblast cell line CHL and hence is not likely to classify as a gene mutant in vitro.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data from study report

Qualifier:

according to guideline

Guideline:

other: as per below mentioned

Principles of method if other than guideline:

Gene mutation assay was conducted to test the potential of test substance.

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium, other: Salmonella strains TA 100 and TA 98

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

10% RLI

Test concentrations with justification for top dose:

0, 1000,2500, 5000,7500, 10000µg/plate

Vehicle / solvent:

Dimethyl Sulfoxide

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

not specified

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; preincubation

Evaluation criteria:

No. of revertants

Statistics:

No data available

Species / strain:

S. typhimurium, other: Salmonella typhimurium strains TA-100 and TA-98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

No data

Remarks on result:

other: No mutagenic effect were observed

Conclusions:

Test substace was considered to be non mutagenic in salmonella strains TA 100 and TA 98 with and without activation.

Executive summary:

Mutagenecity potential of test substance was determined in bacterial strain by AMES assay. The test substance was exposed to salmonella strains TA98 and TA 100 with and without metabolic activation at concentrations of 0,1000,2500, 5000, 7500, 10000, µg/plate. No mutagenic effects were observed in the study both in the presence and absence of metabolic activation. Based on the results obtained the test substance was considered to be non mutagenic with and without activation. Hence the substance cannot be classified as gene mutant in vitro.

Reference 2

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

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 handbook or collection of data

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Principles of method if other than guideline:

A Chromosomal aberration tests in vitro was performed on the test substance to evaluate its mutagenic effect.

GLP compliance:

not specified

Type of assay:

in vitro mammalian chromosome aberration test

Target gene:

No data available

Species / strain / cell type:

mammalian cell line, other: Chinese hamster fibroblast cell line

Details on mammalian cell type (if applicable):

- Type and identity of media: Minimum Essential Medium (MEM; GIBCO) supplemented by 10% calf serum.
- Properly maintained: Yes, by 4 day passages
- Periodically checked for Mycoplasma contamination: No data available
- Periodically checked for karyotype stability: No data available
- Periodically "cleansed" against high spontaneous background: No data available

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data available

Metabolic activation:

not applicable

Test concentrations with justification for top dose:

1,At three different doses with 2 mg/mL being the maximum dose concentration.
2,0.5, 1 and 2 mg/L

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The chemical was soluble in DMSO

Untreated negative controls:

yes

Remarks:

Untreated cells served as negative controls.

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: In medium

DURATION
- Preincubation period: No data available
- Exposure duration: 24 and 48 hrs
- Expression time (cells in growth medium): 24 and 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): Giemsa solution (1.5%, pH 6.8)
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: No data available

NUMBER OF CELLS EVALUATED: 100 well spread metaphases

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

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

Rationale for test conditions:

No data available

Evaluation criteria:

The incidence of polyploid cells as well as of cells with structural chromosomal aberrations such as chromatid or chromosome gaps, breaks, exchanges, ring formations, fragmentations and others, was recorded on each culture plate. In which the incidence of aberrations was usually less than 3.0%. The results were considered to be negative if the incidence was less than 4.9%, equivocal if it was between 5.0 and 9.9%,and positive if it was more than 10.0%.

Statistics:

No data available

Species / strain:

mammalian cell line, other: Chinese hamster fibroblast cell line

Metabolic activation:

not applicable

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

not specified

Species / strain:

other: Chinese hamster fibroblast cell line

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data available
- Effects of osmolality: No data available
- Evaporation from medium: No data available
- Water solubility: No data available
- Precipitation: No data available
- Other confounding effects: No data available

RANGE-FINDING/SCREENING STUDIES: The maximum dose of each sample was selected by a preliminary test in which the dose needed for 50% cell-growth inhibition was estimated using a cell densitometer.

COMPARISON WITH HISTORICAL CONTROL DATA: No data available

ADDITIONAL INFORMATION ON CYTOTOXICITY: No data available

Remarks on result:

other: No mutagenic potential

Conclusions:

The test chemical did not induce chromosomal aberration in the Chinese hamster fibroblast cell line CHL and hence is not likely to classify as a gene mutant in vitro.

Executive summary:

Data for test chemicals was reviewed to determine the mutagenic nature of test chemical. The studies are as mentioned below:

In Chromosomal aberration tests in vitro for test substance in Chinese hamster fibroblast cell line CHL was observed. The cells were exposed to each sample at three different concentration 0.5, 1 and 2 mg/L for 24 and 48 hr. In the present studies, no metabolic activation systems were applied. As the incidence of aberrations 0.0% after 48 hr. The results were considered to be negative .Therefore test substance was considered to be negative in Chromosomal aberration tests in vitro.

 

In Chromosomal aberration tests in vitro for test substance in Chinese hamster fibroblast cell line CHL was observed. The cells were exposed to each sample at three different concentration 0.5, 1 and 2 mg/L for 24 and 48 hr. In the present studies, no metabolic activation systems were applied. As the incidence of aberrations 0.0% after 48 hr. The results were considered to be negative .Therefore test substance was considered to be negative in Chromosomal aberration tests in vitro.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Data for test chemicals was reviewed to determine the mutagenic nature of ethyl formate (109-94-4). The studies are as mentioned below:

In Vitro Genetic Mutation study

AMES Assay

Mutagenicity potential of test substance was determined in bacterial strain by AMES assay. The test substance was exposed to salmonella strains TA98 and TA 100 with and without metabolic activation at concentrations of 0, 1000, 2500, 5000, 7500, 10000, µg/plate. No mutagenic effects were observed in the study both in the presence and absence of metabolic activation. Based on the results obtained the test substance was considered to be non-mutagenic with and without activation. Hence the substance cannot be classified as gene mutant in vitro.

Genetic toxicity of test chemical was tested in Salmonella typhimurium strains TA-1535, TA-1537 and TA-1538 by using plate test (overlay method). The compound was tested at concentration of 1.25, 2.50, and 5.00 % in presence and absence of metabolic activation. DMSO, water/ saline used as solvent control. Ethylmethanesulfonate, 2-Nitrofluorene, Quinacrine mustard used as positive control substance for non activation and 2 Acetylaminofluorene, 8-Aminoquinoline, 2-Aminoanthracene for activation. Rat liver, monkey liver and mouse liver was used as activation system. There were no increase in number of revertants/plate than control with and without Rat liver, monkey liver and mouse liver. Therefore, the test substance is considered to be non mutagenic in Salmonella typhimurium strains TA-1535, TA-1537 and TA-1538 in presence or absence of metabolic activation.

 

Genetic toxicity of test chemical was tested in Salmonella typhimurium strains TA-1535, TA-1537 and TA-1538 by using plate test (overlay method). The compound was tested at concentration of 1.25, 2.50, and 5.00 % in presence and absence of metabolic activation. DMSO, water/ saline used as solvent control. Ethylmethanesulfonate, 2-Nitrofluorene, Quinacrine mustard used as positive control substance for non activation and 2 Acetylaminofluorene, 8-Aminoquinoline, 2-Aminoanthracene for activation. Rat liver, monkey liver and mouse liver was used as activation system. There were no increase in number of revertants/plate than control with and without Rat liver, monkey liver and mouse liver. Therefore, the test chemical is considered to be non mutagenic in Salmonella typhimurium strains TA-1535, TA-1537 and TA-1538 in presence or absence of metabolic activation.

In vitro Chromosomal abbreviation study in mammalian cell

In Chromosomal aberration tests in vitro for test substance in Chinese hamster fibroblast cell line CHL was observed. The cells were exposed to each sample at three different concentration 0.5, 1 and 2 mg/L for 24 and 48 hr. In the present studies, no metabolic activation systems were applied. As the incidence of aberrations 0.0% after 48 hr. The results were considered to be negative .Therefore test substance was considered to be negative in Chromosomal aberration tests in vitro.

 

In Chromosomal aberration tests in vitro for test substance in Chinese hamster fibroblast cell line CHL was observed. The cells were exposed to each sample at three different concentration 0.5, 1 and 2 mg/L for 24 and 48 hr. In the present studies, no metabolic activation systems were applied. As the incidence of aberrations 0.0% after 48 hr. The results were considered to be negative .Therefore test substance was considered to be negative in Chromosomal aberration tests in vitro.

 

Based on the data summarized, ethyl formate (109-94-4)is expected to not induce gene mutation both In Vitro studies .Hence it is not likely to be mutagenic in vitro.

.

Justification for classification or non-classification

Thus based on the above annotation and CLP criteria for target substance ethyl formate (109-94-4)  does not exhibit gene mutation in vitro . Hence the test chemical is not likely to classify as a gene mutant in vitro.