Hexafluoropropene

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Hexafluoropropene

EC Number:

204-127-4

EC Name:

Hexafluoropropene

Cas Number:

116-15-4

Molecular formula:

C3F6

IUPAC Name:

1,1,2,3,3,3-hexafluoroprop-1-ene

Details on test material:

- Purity: 99.998%

Method

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Aroclor-induced rat liver S9

Test concentrations with justification for top dose:

0, 0.026, 0.078, 0.23, 0.69, 2.8, 8.3 and 25 mmoles/L

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

ambient air

Negative solvent / vehicle controls:

no

Positive control substance:

other: 2-aminoanthracene with activation

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

Positive controls:

yes

Positive control substance:

other: 2-nitrofluorene, sodium azide, 9-aminoacridine, and methyl methanesulfonate without activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: The test system was exposed to the test substance via the desiccator methodology, a modification of the plate incorporation methodology.

DURATION: 48-72 hours

NUMBER OF REPLICATIONS: 3

Evaluation criteria:

For each replicate plating, the mean and standard deviation of the number of revertants per plate were calculated and are reported.

For the test substance to be evaluated positive, it must have caused a dose-related increase in the mean revertants per plate of at least one tester strain over a minimum of two increasing concentrations of test substance. Data sets for tester strains TA1535 and TA1537 were judged positive if the increase in mean revertants at the peak of the dose response was greater than or equal to 3.0-times the mean vehicle control value. Data sets for tester strains TA98, TA100 and WP2 uvrA were judged positive if the increase in mean revertants at the peak of the dose response was greater than or equal to 2.0-times the mean vehicle control value.

An equivocal response is a biologically relevant increase in a revertant count that partially meets the criteria for evaluation as positive. This could be a dose-responsive increase that does not achieve the respective threshold cited above or a non-dose responsive increase that is equal to or greater than the respective threshold cited. A response was evaluated as negative, if it is neither positive nor equivocal.

Results and discussion

Test results

Test results 1

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

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

(Without Activation)	Mean Revertants Per Plate
	Strain  TA98	Strain TA100	Strain TA1535	Strain 1537	Strain WP2uvrA
HFP
0 mmole/	12	91	10	4	33
25 mmole/L	0	0	0	0	0
8.3 mmole/L	2	5	0	0	19
2.8 mmole/L	10	59	18	5	20
0.69 mmole/L	8	82	9	6	22
0.23 mmole/L	11	72	12	4	26
0.078 mmole/L	10	98	9	4	23
0.026mmole/L	10	97	10	6	28
Positive control name
2-nitrofluorene (1 µg)	183
sodium azide(1 µg)		901	691
9-Aminoacridine(75 µg)				462
methyl methanesulfonate(1000 µg)					510

 

 

 

 

(With Activation)	Mean Revertants Per Plate
	Strain  TA98	Strain TA100	Strain TA1535	Strain 1537	Strain WP2uvrA
HFP
0 mmole/	15	105	14	6	38
25 mmole/L	0	0	0	0	0
8.3 mmole/L	1	1	0	0	18
2.8 mmole/L	12	71	11	5	25
0.69 mmole/L	16	89	11	3	30
0.23 mmole/L	20	78	11	4	31
0.078 mmole/L	15	93	14	5	31
0.026mmole/L	13	96	11	7	30
Positive control name
2-aminoanthracene (1 µg)	523		208	59
2-aminoanthracene (2 µg)		1762
2-aminoanthracene (10 µg)					241

 

 

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

HFP was not mutagenic in the Ames test.

The study and the conclusions which are drawn from it fulfil the quality criteria (validity, reliability, repeatability).

Executive summary:

The test substance was tested in the Bacterial Reverse Mutation Assay Using Gas-Phase Exposure using Salmonella typhimurium tester strains TA98, TA100, TA1535 and TA1537 and Escherichia coli tester strain WP2uvrA in the presence and absence of Aroclor-induced rat liver S9. The assay was performed in two phases, using the desiccator method, a modification of the plate incorporation method. The first phase, the preliminary toxicity assay, was used to establish the dose-range for the mutagenicity assay. The second phase, the mutagenicity assay was used to evaluate the mutagenic potential of the test substance.

 

Dosing chamber analysis for concentration and stability was not conducted. The interpretation of the study data was based on the nominal dose levels and not on the actual test substance concentration in the exposure chambers as confirmed by analytical analysis. Nevertheless, toxicity in the assay demonstrated that the test system was dosed up to the regulatory-required level. Ambient air was used as the negative control.

In the preliminary toxicity assay (Experiment A1), the dose levels tested were 1.4, 4.2, 6.9, 14, 21, 28 and 37 mmoles/L. No precipitate was observed. Toxicity was observed beginning at 1.4, 4.2, 6.9, 14 or 21 mmoles/L. Based on the findings of the preliminary toxicity assay, the maximum dose plated in the mutagenicity assay was 25 mmoles/L.

 

In the mutagenicity assay (Experiment B1), no positive mutagenic response was observed. The dose levels tested were 0.026, 0.078, 0.23, 0.69, 2.8, 8.3 and 25 mmoles/L. No precipitate was observed. Toxicity was observed beginning at 8.3 or at 25 mmoles/L.

The results of the Bacterial Reverse Mutation Assay Using Gas-Phase Exposure indicate that, under the conditions of this study, the test substance did not exhibit any mutagenic responses in either the presence or absence of Aroclor-induced rat liver S9. Therefore, the test substance was concluded to be negative in this assay.