4-bromo-3,3,4,4-tetrafluorobut-1-ene

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:

March 1977

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

The study was performed pre-GLP and pre-OECD Test Guidelines. It was performed by an experienced reputable laboratory. Limited information is provided on the method used.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1977

Report date:

1977

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

- Principle of test: Analysis of mutagenic activity of 1-Butene, 4-bromo-3,3,4,4-tetrafluoro- in the Salmonella/microsome assay.
- Short description of test conditions: Five histidine-requiring strains of Salomonella typhimurim were sued in the assays. Strains 1535 and TA 100 were used in the detection of base-pair substitution mutations, with strains TA 1537, TA 1538 and TA 98 used ti detect frame-shift mutations. The assays were performed in the presence and absence of a ra-liver homogenate activation system (S.9) .

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

1

Reference substance name:

4-bromo-3,3,4,4-tetrafluorobut-1-ene

EC Number:

242-440-8

EC Name:

4-bromo-3,3,4,4-tetrafluorobut-1-ene

Cas Number:

18599-22-9

Molecular formula:

C4H3BrF4

IUPAC Name:

4-bromo-3,3,4,4-tetrafluorobut-1-ene

Test material form:

not specified

Method

Target gene:

Not stated

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 98

Details on mammalian cell type (if applicable):

Not stated

Additional strain / cell type characteristics:

not specified

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Details on mammalian cell type (if applicable):

Not stated

Additional strain / cell type characteristics:

not specified

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 1537

Details on mammalian cell type (if applicable):

Not stated

Additional strain / cell type characteristics:

not specified

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 100

Details on mammalian cell type (if applicable):

Not stated

Additional strain / cell type characteristics:

not specified

Species / strain 5

Species / strain / cell type:

S. typhimurium TA 1535

Details on mammalian cell type (if applicable):

Not stated

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Type and composition of metabolic activation system:
- source of S9: Not stated
- method of preparation of S9 mix:
- concentration or volume of S9 mix and S9 in the final culture medium: 0.5ml of S.9 mixture was applied to the chemical-top solution. The S.9 mixture contained 0.3 ml of the 9,000 X g supernatant of homognized rat liver
- quality controls of S9 (e.g., enzymatic activity, sterility, metabolic capability): Not stated

Test concentrations with justification for top dose:

The test material was tested at 200, 400, 600, 800, 1000 ug/plate in tests with and without metabolic activity. No justification on test concentrations were provided in the study report

Vehicle / solvent:

- Solvent used: DMSO

- Justification for choice of solvent/vehicle: A justification for the choice of solvent was not provided in the study report.

- Justification for percentage of solvent in the final culture medium: A justification for the percentage solvent in the final culture medium was not provided in the study report.

Controls

Untreated negative controls:

yes

Remarks:

- S.9 control

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

yes

Remarks:

-S.9 control

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

N-ethyl-N-nitro-N-nitrosoguanidine

other: 2- Aminoanthracene

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration: Duplicate
- Number of independent experiments: Two

METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable): Not stated
- Test substance added in medium; A solution of the test material was added to the top agar solution where the solution was then mixed and added to the agar plate. For the metabolic activation system the addition of S.9 mixture was added to the mixture prior to applying to the agar plate.

TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable: N/A
- Exposure duration/duration of treatment:49 hours
- Harvest time after the end of treatment (sampling/recovery times): Not stated

METHODS FOR MEASUREMENTS OF GENOTOXICIY
Not stated

Rationale for test conditions:

A rationale for the test conditions was not provided in the study report.

Evaluation criteria:

The criteria used to determine the mutagenic potential of the test substance was a significant/lack of increase in the spontaneous mutation frequency.

Statistics:

No statistics are provided in the study report.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table 1: Mutagenic activity of 1-Butene, 4-Bromo-3,3,4,4-Tetrafluoro- in Salmonella Typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100 with metabolic activation

 

			Histidine+ Revertants Per Plate **
Compound added			TA 1535	TA 1537	TA 1538	TA 98	TA 100
DMSO			13	17	21	37	101
-S.9*	(1000 µg/plate)		12	10	7	18	100
10, 943	200 µg/plate		7	11	18	37	98
	400 µg/plate		9	17	20	35	98
	600 µg/plate		12	20	22	42	118
	800 µg/plate		12	14	21	45	98
	1000 µg/plate		13	13	14	29	100
2AA	5 µg/plate		-	-	-	-	933
	10 µg/plate		209	-	782	2561	-
	100 µg/plate		-	284	-	-	-

DMSO = Dimethylsulfoxide (solvent control)

10,943 = 1-butene, 4-bromo-3,3,4,4-tetrafluoro-

2AA = 2-Aminoanthracene (positive control)

* = Test plate without S.9 Activation

** = Average of two plates

 

Table 2: Mutagenic activity of 1-Butene, 4-Bromo-3,3,4,4-Tetrafluoro- in Salmonella Typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100 without metabolic activation

 

			Histidine+ Revertants Per Plate *
Compound added			TA 1535	TA 1537	TA 1538	TA 98	TA 100
DMSO			4	14	7	22	93
10, 943	200 µg/plate		9	16	13	20	91
	400 µg/plate		6	23	15	29	83
	600 µg/plate		9	19	9	22	95
	800 µg/plate		9	15	11	26	78
	1000 µg/plate		5	15	9	26	101
MNNG	2 µg/plate		1966		-	-	1278
9AAc	1050 µg/plate		-	2022	--	-	-
2NF	10025 µg/plate		-	-	1964	3915	-

DMSO = Dimethylsulfoxide (solvent control)

10,943 = 1-butene, 4-bromo-3,3,4,4-tetrafluoro-

MNNG = N-Methyl-N’-nitro-N-nitrosoguanidine (positive control)

9AAc = 9-Aminoacridine

2NF = 2-ANitrofluorene (positive control)

* = Average of two plates

Applicant's summary and conclusion

Conclusions:

Under the conditions of this study, the test item is considered not mutagenic in concentrations up to 1000 ug/plate in either the presence or absence of a liver microsomal system on Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100.

Executive summary:

The study was performed to assess the mutagenic potential of the test material to Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100 in the presence and absence of a liver microsomal system (S.9). For treatments in the absence of the liver microsomal system a volume of 0.1mL test substance and 10⁸ bacterial were added to 2ml of the top agar. This solution was applied to the surface of the Davis minimal agar plate.

 

Treatment in the presence of the liver microsomal system were the same as that in the absence of the liver microsomal system with the exception that 0.5 mL S.9. mixture was added to the chemical topped agar solution. All plates were incubated at 37˚C for 48 hours. Positive and negative controls were used in both test systems and it was not indicated if these were within the acceptable historical control ranges. No applicability criteria were provided for this test method therefore this could not be determined. Under the conditions of the study, the test substance showed no evidence of mutagenic activity and it was concluded to be negative as it did not significantly increase the spontaneous mutation frequency in Salmonella typhimurium.