2,2,3,3-Tetrafluorooxetane

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 November 2009 to 7 December 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test conducted in compliance with recognised guidelines and to GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries. Test Data for Registration of Agricultural Chemicals, 12 Nohsan No. 8147, Guideline 2-1-19-1, Agricultural Production Bureau, November 24, 2000.

Deviations:

no

Test guideline 5

Qualifier:

according to guideline

Guideline:

other: ICH (1996) Guideline S2A: Guidance on Specific Aspects of Regulatory Genotoxicity Tests for Pharmaceuticals. PAB/PCD Notification No. 444.

Deviations:

no

Principles of method if other than guideline:

other guidelines quoted in the report are:

Official notice of J MHLW, METI and ME (21 November 2003):
YAKUSHOKUHATSU No.1121002
SEIKYOKU No.2
KANPOKIHATSU No. 031121002

Official Notice of J MOL (8 February 1999)

ICH (1998) Guideline S2B: Genotoxicity: A Standard Battery for Genotoxicity Testing of Pharmaceuticals. PMSB/ELD Notification No. 554.

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,2,3,3-tetrafluorooxetane

EC Number:

700-202-5

Cas Number:

765-63-9

Molecular formula:

C3H2F4O

IUPAC Name:

2,2,3,3-tetrafluorooxetane

Details on test material:

- Name of test material (as cited in study report): 2,2,3,3-tetrafluorooxetane (TFO)
- Physical state: Transparent liquid
- Analytical purity: 99%
- Purity test date: no data given
- Lot/batch No.: TFOS285001
- Expiration date of the lot/batch: 30 April 2010
- Stability under test conditions: no data given
- Storage condition of test material: Room temperature

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Additional strain / cell type characteristics:

not applicable

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A pKM 101

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Rat liver homogenate fraction (S9 mix)

Test concentrations with justification for top dose:

5-5000 µg per plate of TFO (test 1)
5-500 µg per plate of TFO (test 2)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: TFO not miscible with water but miscible with DMSO, therefore DMSO used.

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

DMSO

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Concentration 2 µg per plate

Migrated to IUCLID6: TA100 & TA1535 without S9 mix

Controls 2

Positive control substance:

9-aminoacridine

Remarks:

Concentration 50 µg per plate

Migrated to IUCLID6: TA1537 without S9 mix

Controls 3

Positive control substance:

2-nitrofluorene

Remarks:

Concentration 2 µg per plate

Migrated to IUCLID6: TA98 without S9 mix

Controls 4

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

Concentration 2 µg per plate

Migrated to IUCLID6: WP2 uvrA (pKM101) without S9 mix

Controls 5

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene, TA100, TA1535 & WP2 uvrA (pKM101) with S9 mix

Remarks:

Concentration 5 µg per plate for TA100, TA1535, 10 µg per plate for WP2 uvrA (pKM101)

Controls 6

Positive control substance:

benzo(a)pyrene

Remarks:

Concentration 5 µg per plate

Migrated to IUCLID6: TA98 & TA1537 with S9 mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 30 minutes
- Exposure duration: 72 hours

Concentrations of TFO up to 5000 µg/plate were tested initially. This is the standard limit concentration recommended in the regulatory guidelines that this assay follows. Other concentrations used were a series of ca half-log10 dilutions of the highest concentration. A lower maximum concentration was tested in some strains in the second test, in order to avoid excessive toxicity.

Evaluation criteria:

If exposure to a test substance produces a reproducible increase in revertant colony numbers of at least twice (three times in the case of strains TA1535 and TA1537) the concurrent vehicle controls, with some evidence of a positive dose-response relationship, it is considered to exhibit mutagenic activity in this test system.
If exposure to a test substance does not produce a reproducible increase in revertant colony numbers, it is considered to show no evidence of mutagenic activity in this test system.

Statistics:

No statistical analysis was performed.

Results and discussion

Test results

Test results 1

Species / strain:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

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:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

COMPARISON WITH HISTORICAL CONTROL DATA:

The mean revertant colony counts for the vehicle controls were within or close to the 99% confidence limits of the current historical control range of the laboratory. (For tabulated historical data, see attachment below).

Appropriate positive control chemicals (with S9 mix where required) induced substantial increases in revertant colony numbers with all strains in all reported tests, confirming sensitivity of the cultures and activity of the S9 mix.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

The absence of colonies on sterility check plates confirmed the absence of microbial contamination of the S9 mix, buffer and test substance formulation.

The viability counts confirmed that the viable cell density of the cultures of the individual organisms exceeded 10⁹/mL in all cases, and therefore met the acceptance criteria.

For results tables, see the attachments given below.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive

TFO exhibited mutagenic activity in this bacterial system under the test conditions employed. Activity was enhanced following metabolic activation.