2,3,3,4,4-pentafluoro-5-methoxy-2,5-bis[1,2,2,2-tetrafluoro-1-(trifluoromethyl)ethyl]tetrahydrofuran

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1997

Deviations:

yes

Remarks:

In the pre-incubation assay, 50 uL of dilution of the test item in ethanol was added to the top agar. The amount of vehicle was determined to have no influence on the results of the pre-incubation assay.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,3,3,4,4-pentafluoro-2,5-bis(1,1,1,2,3,3,3-heptafluoropropan-2-yl)-5-methoxyoxolane

EC Number:

812-244-2

Cas Number:

957209-18-6

Molecular formula:

C11H3F19O2

IUPAC Name:

2,3,3,4,4-pentafluoro-2,5-bis(1,1,1,2,3,3,3-heptafluoropropan-2-yl)-5-methoxyoxolane

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Lot # 20003, unit 0022
- Expiration date of the lot/batch: 30 March, 2018
- Purity test date: 30 March, 2016

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test article was dissolved in ethanol.

FORM AS APPLIED IN THE TEST: The test article was applied using ethanol as a vehicle.

Method

Target gene:

Histidine operon, tryptophan operon.

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

Arcolor 1254-induced rat liver S9 homogenate

Test concentrations with justification for top dose:

52, 164, 512, 1600, 5000 ug/plate. Top dose was chosen based on a range-finding assay.

Vehicle / solvent:

Ethanol

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

2-nitrofluorene

sodium azide

methylmethanesulfonate

other: ICR-191, 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION:in agar (plate incorporation) and a preincubation assay
- Cell density at seeding (if applicable): 10^9 cells/mL

DURATION
- Preincubation period: In agar: None, Pre-incubation assay: 30 minutes
- Exposure duration: In agar: 48 hours, Pre-incubation assay: 48 hours
- Expression time (cells in growth medium): 48.5 and 48 hours.
- Selection time (if incubation with a selection agent): 48.5 and 48 hours
- Fixation time (start of exposure up to fixation or harvest of cells): 48 hours

SELECTION AGENT (mutation assays): Agar with a limited amount of histidine or tryptophan.

NUMBER OF CELLS EVALUATED: Revertant colonies were counted automatically with the Sorcerer Colony Counter.

DETERMINATION OF CYTOTOXICITY
- Method: Bacterial background lawn health.

Rationale for test conditions:

The test was conducted according to OECD Guideline 471.

Evaluation criteria:

A test article is considered negative (not mutagenic) if:
-The total number of revertants in tester strain TA100 or WP2uvrA is not greater than two times the concurrent control, and the total number of revertants in the tester strains TA1535, TA1537 or TA98 is not greater than three times the concurrent control.
-The negative response should be reproducible in at least one follow up experiment.

A test article is considered positive (mutagenic) if:
-The total number of revertants in the tester strain TA100 or WP2uvrA is greater than two times the concurrent control, or the number of revertants in tester strains TA1535, TA1537 or TA98 is greater than three times the concurrent control.
-In case a repeat experiment is performed when a positive response is observed in one of the tester strains, the positive response should be reproducible in at least one follow up experiment.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation of the test article was observed at the beginning of the incubation period at 5000 ug/plate. At the end of the incubation period, no precipitation was observed.
- Definition of acceptable cells for analysis:
- Other confounding effects:

RANGE-FINDING/SCREENING STUDIES: The test article was tested up to 5000 ug/plate.

Applicant's summary and conclusion

Conclusions:

Based on the results of the study, the test article is not mutagenic in the Ames assay in the presence or absence of metabolic activation.

Executive summary:

The mutagenicity potential of the test article was evaluation in the Salmonella typhimurium (TA1535, TA1537, TA100 and TA98) reverse mutation assay and the Escherichia coli (WP2uvrA) reverse mutation assay using the plate incorporation and pre-incubation methods. The study was conducted according to OECD Guideline 471 (1997) in compliance with OECD GLP (1997) regulations. The assay was performed in two independent experiments, at first a direct plate assay was performed and secondly a pre-incubation assay both in the absence and presence of S9 -mix (Aroclor 1254 -induced rat liver S9 mix). The test article was prepared in ethanol and tested at 52, 164, 512, 1600, 5000 ug/plate in each assay. Doses were selected based on a range-finding assay. Precipitation of the test article was observed at the beginning of the incubation period at 5000 ug/plate.  At the end of the incubation period, no precipitation was observed. The test article did not induce a significant dose-related increase in the number of revertant colonies in any of the strains tested. Based on the results of the study, the test article is not mutagenic in the Ames assay in the presence or absence of metabolic activation.