[No public or meaningful name is available]

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05/Oct/98 - 27/Jan/99

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosomal Aberration Test)

Deviations:

no

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

1,1,2,2-tetrafluoroethyl-2,2,2-trifluoroethyl ether

EC Number:

609-858-6

Cas Number:

406-78-0

Molecular formula:

C4H3F7O

IUPAC Name:

1,1,2,2-tetrafluoroethyl-2,2,2-trifluoroethyl ether

Test material form:

liquid

Specific details on test material used for the study:

Name: 1,1,2,2-tetrafluoroethyl-2,2,2-trifluoroethyl ether [HFE 347 pc-f]
BML test substance number: BML-4262
Lot number: T52068101
Purity: 99.8wt%
Impurity name and concentration: No information
Appearance at room temperature: Liquid
Stability: Stable at 120°C under the presence of iron or SUS for 3 days
Solubility: Water (0.011g/100g H20), DMSO (>1000mg/mL), Acetone (Readily soluble)
Stability in solvent: Water (stable, no change), DMSO (stable, no change, dissolution in 1:1 mixture), Acetone (stable, no change, dissolution in 1:1 mixture)
Storage condition: Room temperature

Method

Species / strain

Species / strain / cell type:

Chinese hamster lung (CHL/IU)

Metabolic activation:

with and without

Metabolic activation system:

Composition of S9 mix (in 10mL): S9 (3mL), Cofactor (7mL), HEPES buffer (40umol), MaCl2 (50umol), KCl (330umol), G-6-P (50umol), NADP (40umol)

Test concentrations with justification for top dose:

The dose range in the chromosome aberration test was selected in accordance with the results of the cell growth inhibition test. From the results of the cell growth inhibition test, cell growth inhibition in both short-term and continuous exposure experiments were not observed at 5.0mg/L (the maximum concentration). Based on this result, the maximum concentration of test substance in 24 and 48 hour exposure experiments was selected at 5.0mg/mL, with 3 lower concentrations diluted from the maximum concentration by a common ratio of 2.

Vehicle / solvent:

The solvent selected was Dimethyl sulfoxide (DMSO) since the test substance had a low solubility in water at (0.011g/100g)

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

mitomycin C

other: Physiological saline

Details on test system and experimental conditions:

Cultured Mammalian cells: The cell line of the Chinese hamster fibroblasts in lung (CHL/IU cells, modal chromosome number of 25) was used.
Cell culture medium: Eagle's MEM supplemented with 10% calf serum was used.
Growth form: The cell cultures were incubated using a CO2 gas incubator under the conditions of a highly humidified atmosphere with 5% CO2 at 37°C.
Cell stock conditions: Cells were stored in the deep temperature freezer (-80°C) or tank with liquid nitrogen (-196°C). 10v/v% of dimethyl sulfoxide was added to the culture medium at the time of storage.

Results and discussion

Test results

Key result

Species / strain:

Chinese hamster lung (CHL/IU)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Additional information on results:

In the results of the chromosome aberration test in the short-term exposure experiment both with and without the S9 mix, the frequency of cells carrying structural chromosome aberrations or polyploidy were not increased at any concentration compared with negative (untreated and solvent) controls. In the positive (MMC) control group, in the short-term exposure experiment without S9 mix, the frequency of cells carrying structural chromosome aberrations was 16%. In the positive (DMN) control group, in the short-term exposure experiment with S9 mix, the frequency of cells carrying structural chromosomal aberrations was 70%.

In the results of the chromosome aberration test in both 24- and 48-hour exposure, the frequency of cells carrying structural chromosome aberrations or polyploidy were not increased at any concentration compared with negative controls. In the positive (MMC) control groups in continuous exposure experiments, the frequencies of cells carrying structural chromosome aberrations in 24 and 48 hour exposure were 39.5 and 46.5% respectively.

Applicant's summary and conclusion

Conclusions:

Under the test conditions described, it was concluded that 1,1,2,2-tetrafluoroethyl-2,2,2-trifluoroethyl ether did not induce chromosome aberrations in the cell line of Chinese hamster fibroblast in the lung (CHL/IU cells).

Executive summary:

An in vitro chromosome aberration test in cultured mammalian cells (CHL/IU cells) was performed to assess the potential of 1,1,2,2-tetrafluoroethyl-2,2,2-trifluoroethyl ether to induce chromosome aberrations.
Since cell growth inhibition in both short-term and continuous exposure experiments were not observed at 5.0mg/mL (max concentration) in the results of the cell growth inhibition test, the concentrations of 50% cell growth inhibition for this test substance were more than 5.0mg/mL. Therefore, the maximum concentration of test substance in short term and continuous exposure experiments was 5.0mg/mL, and 3 applied concentrations which were prepared with dilution by a common ratio of 2 were set up and examined.

In the results of the chromosome aberration test in short-term and continuous exposure experiments, the frequency of cells carrying structural chromosome aberrations or polyploidy were not increased at any concentration compared with negative (untreated and solvent) controls.

Based on the above results, it was concluded that 1,1,2,2-tetrafluoroethyl-2,2,2-trifluoroethyl ether did not induce chromosomal aberrations in CHL/IU cells.