2-(1,2-dichloro-1,2,2-trifluoroethoxy)-1,1,2,2-tetrafluoroethanesulfonyl fluoride

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1996

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

Study predates the OECD guidelines

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Qualifier:

no guideline followed

Version / remarks:

Study predates the OECD guideline 471. Although the protocol used seems to be similar to the OECD guideline, the lack of information concerning the test conditions does not allow to conclude that they are formally similar.
Deviations from OECD 471:Only 4 strains tested / duplicate plating / no follow-up experiment.

Principles of method if other than guideline:

Mutagenicity in the Salmonella typhimurium plate incorporation assay (Reverse mutation assay):

The test substance was evaluated for mutagenicity in Salmonella typhimurium strains TA100, TA1535, TA97a and TA98 with and without an exogenous metabolic activation system (S9). The maximum concentration tested was 5000 µg/plate. Concentrations of 2500, 1000, 500, 100, 50 and 10 µg/plate were also tested in comparison to a negative (solvent) control. The negative and the diluent for the test substance was ethyl alcohol.

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-(1,2-dichloro-1,2,2-trifluoroethoxy)-1,1,2,2-tetrafluoroethane-1-sulfonyl fluoride

EC Number:

604-439-4

Cas Number:

144728-59-6

Molecular formula:

C4Cl2F8O3S

IUPAC Name:

2-(1,2-dichloro-1,2,2-trifluoroethoxy)-1,1,2,2-tetrafluoroethane-1-sulfonyl fluoride

Test material form:

liquid

Details on test material:

See confidential details on test material

Method

Target gene:

Not specified.

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA100, TA1535, TA97a and TA98

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

The maximum concentration tested was 5000 µg/plate.
Concentrations of 2500, 1000, 500, 100, 50 and 10 µg/plate were also tested.

Vehicle / solvent:

ethyl alcohol

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

ethyl alcohol

Positive controls:

not specified

Controls 2

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene (2AA)

Remarks:

With metabolic activation

Controls 3

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Without metabolic activation

Controls 4

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

Without metabolic activation

Controls 5

Positive controls:

yes

Positive control substance:

other: ICR-191

Remarks:

Without metabolic activation

Details on test system and experimental conditions:

Not specified.
No details provided in the report.

Rationale for test conditions:

No information on the test conditions

Evaluation criteria:

According to the test method, the test substance was classified as positive when:
1. The average number of revertants in any strain at any test substance concentration studied was at least two times greater than the average number of revertants in the concurrent negative control.
AND
2. There was a positive dose response relationship in that same strain.

The test substance was classified as negative when:
1. Ther were no test concentrations with an average number of revertants at least two times greater than the average number of revertants in the negative control.
OR
2. There was a positive dose-response relationship.

A test substance was classified as equivocal when the test substance was not clearly negative yet did not meet the criteria for a positive response.

Statistics:

For each strain, the average of revertants and the standard deviation (S.D.) at each concentration with and without S9 activation were calculated.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Background lawn was noticeably thinner and/or size of microcolonies was slightly larger than control at 50 microgram/plate both in presence and absence of S9-mix. More evident signs of cytotoxicity were observed at higher concentrations.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

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:

Background lawn was noticeably thinner and/or size of microcolonies was slightly larger than control at 500 microgram/plate (S9mix -) and 100 (S9mix +). More evident signs of cytotoxicity were observed at higher concentrations.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

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:

cytotoxicity

Remarks:

Background lawn was noticeably thinner and/or size of microcolonies was slightly larger than control at 50 microgram/plate both in presence and absence of S9-mix. More evident signs of cytotoxicity were observed at higher concentrations.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 97

Remarks:

TA97a strain

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Background lawn was noticeably thinner and/or size of microcolonies was slightly larger than control at 100 microgram/plate both in presence and absence of S9-mix. More evident signs of cytotoxicity were observed at higher concentrations.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

According to the test method, acceptability criteria:
An individual trial must have included at least five test concentrations, a negative controm, and a positive indicator for each selected tester strain. Data acceptability criteria were as follows:
- A single data point may have been rejected if contamination or excessive toxicity was seen on a treatment plate. A single data point may also have been rejected if excessive precipitate on the plate prevented accurate colony counting.
- A negative control data point may have been rejected if it fell outside the acceptable spontaneous mutation range.
- A concentration level was rejected if there were less than two data points at the treatment level or if the data point values were judged by the study director to be too divergent.
- A trial for the affected strain was rejected if the negative control was rejected or if there was no evidence of mutagenic activity on any positive indicator plate.

Only those trials which met the criteria of acceptability were included in the report.

Any other information on results incl. tables

Table 1. Mutagentic activity of the test item in strain TA100

	Without Metabolic Activation			With Metabolic Activation
Concentration (µg/plate)	Revertant average (SD)	Observations	Concentration (µg/plate)	Revertant average (SD)	Observations
0.0	94 (6)	T0, P0	0.0	125 (4)	T0, P0
10.0	88 (4)	T0, P0	10.0	106 (4)	T0, P0
50.0	78 (4)	T1, P0	50.0	92 (6)	T1, P0
100.0	80 (3)	T1, P0	100.0	102 (1)	T1, P0
500.0	87 (2)	T1, P0	500.0	110 (6)	T2, P0
1000.0	70 (0)	T3, P0	1000.0	100 (12)	T1, P0
2500.0	61 (1)	T3, P0	2500.0	94 (5)	T3, P1
5000.0	75 (1)	T3, P0	5000.0	90 (1)	T3, P1
Positive control: NAAZ			Positive control: 2AA
2 µg/plate	610 (23)	N	1 µg/plate	2341 (306)	N

Table 2. Mutagentic activity of the test item in strain TA1535

	Without Metabolic Activation			With Metabolic Activation
Concentration (µg/plate)	Revertant average (SD)	Observations	Concentration (µg/plate)	Revertant average (SD)	Observations
0.0	20 (1)	T0, P0	0.0	25 (1)	T0, P0
10.0	18 (2)	T0, P0	10.0	22 (1)	T0, P0
50.0	21 (1)	T0, P0	50.0	20 (4)	T0, P0
100.0	19 (8)	T0, P0	100.0	18 (1)	T1, P0
500.0	15 (1)	T1, P0	500.0	19 (2)	T1, P0
1000.0	15 (1)	T2, P0	1000.0	15 (1)	T2, P0
2500.0	20 (1)	T2, P0	2500.0	9 (1)	T2, P0
5000.0	17 (1)	T3, P0	5000.0	7 (2)	T2, P1
Positive control: NAAZ			Positive control: 2AA
2 µg/plate	547 (4)	N	2 µg/plate	536 (23)	N

Table 3. Mutagentic activity of the test item in strain TA98

	Without Metabolic Activation			With Metabolic Activation
Concentration (µg/plate)	Revertant average (SD)	Observations	Concentration (µg/plate)	Revertant average (SD)	Observations
0.0	34 (2)	T0, P0	0.0	21 (1)	T0, P0
10.0	23 (1)	T0, P0	10.0	28 (2)	T0, P0
50.0	24 (2)	T1, P0	50.0	28 (2)	T1, P0
100.0	23 (1)	T1, P0	100.0	20 (1)	T1, P0
500.0	19 (2)	T1, P0	500.0	26 (2)	T1, P0
1000.0	18 (2)	T2, P0	1000.0	30 (1)	T2, P0
2500.0	6 (1)	T3, P0	2500.0	17 (2)	T3, P0
5000.0	9 (1)	T3, P0	5000.0	18 (1)	T3, P0
Positive control: 2NF			Positive control: 2AA
25 µg/plate	1710 (31)	N	2 µg/plate	2048 (214)	N

Table 4. Mutagentic activity of the test item in strain TA97a

	Without Metabolic Activation			With Metabolic Activation
Concentration (µg/plate)	Revertant average (SD)	Observations	Concentration (µg/plate)	Revertant average (SD)	Observations
0.0	89 (3)	T0, P0	0.0	98 (3)	T0, P0
10.0	108 (0)	T0, P0	10.0	84 (7)	T0, P0
50.0	65 (1)	T0, P0	50.0	69 (10)	T0, P0
100.0	48 (6)	T1, P0	100.0	78 (2)	T1, P0
500.0	63 (9)	T2, P0	500.0	85 (6)	T1, P0
1000.0	71 (0)	T2, P0	1000.0	92 (1)	T2, P0
2500.0	72 (4)	T2, P0	2500.0	105 (5)	T2, P0
5000.0	55 (1)	T2, P0	5000.0	67 (5)	T3, P0
Positive control: ICR-191			Positive control: 2AA
2 µg/plate	2326 (45)	N	1 µg/plate	1491 (1)	N

Abbreviations used in the tables:

Evidence for test substance toxicity to the bacteria was documented by recording the appearance of the plates and background lawn using the following key:

- T0: Background lawn appeared normal.

- T1: Background lawn was noticeably thinner and/or size of micrcolonies was slightly larger than controls.

- T2: Background lawn was markedly thinner and/or size of micrcolonies was markedly larger than controls.

- T3: Background lawns were severely reduced and/or micrcolonies were greatly enlarged relative to controls.

- T4: Background lawn was absent and/or microcolonies could be seen readily by the unaided eye.

- T5: Colony formation was reduced or absent relative to controls.

Formation of a precipitate by the test material was documented using the following key:

- P0: No evidence of precipitate was detected.

- P1: Microscopic precipitate was present.

- P2: Marked precipitate was present but did not interfere with automated colony counting.

- P3: Marked precipitate was present and required the plate to be counted by hand.

- P4: Heavy precipitate prevented accurate colony counting and/or obscured the background lawn.

- N: The absence of any noteworthy observations.

- R: Plate rejected.

Applicant's summary and conclusion

Conclusions:

Under the conditions of this study, no evidence of mutagenic activity was detected.

Executive summary:

The assessment of the mutagenic activity of the test substance was carried out using the Salmonella typhimurium reverse mutation assay (plate incorporation method).

The test substance was tested in the Salmonella typhimurium reverse mutation assay with four histidine-requiring strains of Salmonella typhimurium (TA100, TA1535, TA97a and TA98). The test was performed in the absence and presence of S9 -mix.

Under the conditions of this study Fluorosulfonic Adduct did not induce a significant dose-related increase in the number of revertant (His+ ) colonies in each of the four tester strains (TA1535, TA97a, TA98 and TA100) both in the absence and presence of S9-metabolic activation.

Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies.