Butane, 1,2,3,4-tetrachloro-1,1,2,3,4,4-hexafluoro-

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:

28 january 2003 to 10 june 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,2,3,4-tetrachloro-1,1,2,3,4,4-hexafluorobutane

EC Number:

938-148-1

Cas Number:

375-45-1

Molecular formula:

C4Cl4F6

IUPAC Name:

1,2,3,4-tetrachloro-1,1,2,3,4,4-hexafluorobutane

Details on test material:

- Name of test material (as cited in study report): A1112 Dimer
- Substance type: pure substance
- Physical state: liquid
- Analytical purity: 99%
- Impurities (identity and concentrations): not reported
- Composition of test material, percentage of components: 99% pure substance
- Isomers composition: not applicable
- Purity test date: not reported
- Lot/batch No.: DAD 619
- Expiration date of the lot/batch: 2005

- Storage condition of test material: ambient temperature in the dark

Method

Target gene:

Salmonella typhimuriumtester strains: histidine requirement
E. coli (WP2 uvrA): tryptophan requirement

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

other: uvrB deletion (S. typhimurium), uvrA mutation (E.coli)

Metabolic activation:

with and without

Metabolic activation system:

rat liver S9-mix

Test concentrations with justification for top dose:

Test item: 6 doses per strain (5000, 2500, 1000, 500, 200, 100)microgram/plate, 3 plates per dose.
Positive controls: 1 dose per strain, 3 plate per dose.

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

N-ethyl-N-nitro-N-nitrosoguanidine

benzo(a)pyrene

other: see remarks

Rationale for test conditions:

test concentrations were selected up to the recommended limit.

Evaluation criteria:

Validity criteria:
- the concurrent solvent control data are acceptable
- the positive control data show acceptable increases

Interpretation criteria in the study:
Positive response when one or both criteria are met:
- a significant, dose-related increase in the mean number of revertants
- a 2-fold or greater increase in the mean number of revertant colonies (over that of the concurrent solvent control plates), at one or more concentrations

Negative response when:
- no significant dose-related increase in the mean number of revertant colonies per plate and,
- in the absence of any such response, no increase in colony number (at any concentration) which exceeds 2x the concurrent solvent control.

Statistics:

no

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

reduced number of colonies in the pre-incubation experiment with S9

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

reduced number of colonies in the pre-incubation experiment with S9

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Species / strain:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Under the test conditions, 1,2,3,4-tetrachloro-hexafluorobutane gave a negative, i.e. non mutagenic response in S. typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 and E. coli starin WP2P uvrA in both the presence and absence of S9-mix.

Executive summary:

1,2,3,4-tetrachloro-hexafluorobutane was evaluated in a bacterial mutagenicity assay (based on Maron and Ames (1983)) over a range of concentrations using four strains of Salmonella typhimurium ( TA 1535, TA 1537, TA 98 and TA 100) and one strain of Escherichia coli (WP2P uvrA) in the presence and absence of a rat liver - derived metabolic activation system (S9 -mix).

In two separate experiments, the test substance did not induce any significant, reproducible increases in the observed numbers of revertant colonies in any of the tester strains, either in the presence or absence of S9 -mix.

The sensitivity of the test system, and the metabolic activity of the S9-mix, were clearly demonstrated by the increases in the numbers of revertant colonies induced by positive control substances.

It is concluded that, under the conditions of this assay, 1,2,3,4-tetrachloro-hexafluorobutane gave a negative, i.e. non mutagenic response in S. typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 and E. coli starin WP2P uvrA in both the presence and absence of S9-mix.