Dichloromethyl(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)silane

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:

2002-05-03 to 2002-08-01

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Dichloromethyl(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)silane

EC Number:

277-551-0

EC Name:

Dichloromethyl(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)silane

Cas Number:

73609-36-6

Molecular formula:

C9H7Cl2F13Si

IUPAC Name:

(dichloromethyl)(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)silane

Constituent 2

Reference substance name:

[2-(perfluorohexyl)ethyl]dichloro(methyl)silane

IUPAC Name:

[2-(perfluorohexyl)ethyl]dichloro(methyl)silane

Test material form:

other: liquid

Method

Target gene:

histidine operon

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

Aroclor induced rat liver S9

Test concentrations with justification for top dose:

10, 31.6, 100, 316, 1000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Ethylene glycol dimethylether

- Justification for choice of solvent/vehicle: Solubility properties and relative non-toxicity to bacteria

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA 1535, TA 100 (without activation)

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA 98 (without activation)

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA 1537 (without activation)

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

TA 102 (without activation)

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-anthracene amide

Remarks:

TA 98, TA 102, TA 1537 (with activation)

Controls 6

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

TA 100, TA 1535 (with activation)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium (plate incorporation); preincubation

DURATION

- Preincubation period: 60 minutes

- Expression time (cells in growth medium): 48 hours

NUMBER OF REPLICATIONS: 3 plates for each test concentration; the initial plate incorporation assay was repeated using the pre-incubation method.

DETERMINATION OF CYTOTOXICITY

- Method: Background lawn assessment, relative colony counts

METABOLIC ACTIVATION
Aroclor induced rat liver S9 was tested for protein and P-450 content. S9 mix contained 5% S9, and glucose-6-phosphate and NADP as co-factors. 0.5 ml S9 mix were added to 2 ml top agar, 0.1 ml test material and 0.1 ml cell suspension, giving a final concentration of approximately 1% S9.

Evaluation criteria:

A result is positive if the number of revertants is significantly increased compared with the solvent control to at least 2-fold of the solvent control for TA 98, TA 100 and TA 102 and 3-fold of the solvent control for TA 1535 and TA 1537 in both experiments.

Positive results have to be reproducible and the histidine independence of the revertants has to be confirmed by streaking on histidine-free agar plates.

Cytotoxicity is defined as a reduction in the number of colonies by >50% compared with the solvent control and/or a sparse background lawn.

Statistics:

MANN and WHITNEY and Spearman’s rank.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

316 - 1000 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

1000 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

1000 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

316 - 1000 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

316 - 1000 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 6

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

1000 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 7

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

316 - 1000 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

COMPARISON WITH HISTORICAL CONTROL DATA: Results were within range of historical control data

Any other information on results incl. tables

Table 2: Dose range-finding study Number of revertants per plate (mean of 2 plates)

	TA100
Conc. (µg/plate)	Plate 1	Plate 2	Cytotoxic (yes/no)
0*	112	115	No
0.316	117	105	No
1	111	110	No
3.16	126	112	No
10	101	114	No
31.6	116	121	No
100	172	181	No
316	165	173	No
1000	132	154	Yes
3160	0	0	Yes
5000	0	0	Yes

*solvent control with Ethylene glycol dimethylether

Table 3a: Experiment 1 Plate incorporation Number of revertants per plate (mean of 3 plates)

	TA98			TA100			TA102
Conc. (µg/plate)	— MA	+ MA	Cytotoxic (yes/no)	—  MA	+ MA	Cytotoxic (yes/no)	— MA	+  MA	Cytotoxic (yes/no)
0*	42	51.7	No	133	158	No	278	295	No
10	36.3	48.7	No	154	148	No	274.7	273.7	No
31.6	34.7	53	No	139	153.3	No	268.3	275	No
100	34.3	57	No	137.7	131.3	No	271.3	270	No
316	47.7	47.3	No	143.3	139.3	No	258	272.3	No
1000	31.3	40.3	Yes	131	160.7	Yes	286.7	267.3	Yes
Positive control	1145.7	1100	No	1194.3	1196.3	No	1320	1150.3	No

*solvent control with Ethylene glycol dimethylether

Table 3b: Experiment 1 Plate incorporation Number of revertants per plate (mean of 3 plates)

	TA1535			TA1537
Conc.µg/plate	— MA	+  MA	Cytotoxic (yes/no)	— MA	+  MA	Cytotoxic (yes/no)
0*	13	15.3	No	6	4	No
10	13	12	No	3.7	4.7	No
31.6	14.7	14	No	3.3	5.7	No
100	14.3	13	No	3	2.7	No
316	11	13	No	3.7	4.7	No
1000	12	16.3	No	4.3	3.3	No
Positive control	1187	1189	No	1191.3	1241.7	No

*solvent control with Ethylene glycol dimethylether

Table 4a: Experiment 2 Preincubation Number of revertants per plate (mean of 3 plates)

	TA98			TA100			TA102
Conc. µg/plate	— MA	+  MA	Cytotoxic (yes/no)	— MA	+  MA	Cytotoxic (yes/no)	—  MA	+  MA	Cytotoxic (yes/no)
0*	37.3	30.7	No	143.7	154	No	280.3	290.3	No
10	31.7	30.3	No	187	160.7	No	276.3	283.7	No
31.6	38	35.7	No	171.7	144.7	No	288.7	286.3	No
100	35	36.3	No	178	152	No	283.3	281.7	No
316	0	36	Yes	162.7	144.7	No	297.3	265	Yes
1000	0	0	Yes	0	0	Yes	0	0	Yes
Positive control	894.3	983.7	No	1326.3	1333.3	No	1338.3	1344.3	No

*solvent control with Ethylene glycol dimethylether

Table 4b: Experiment 2 Preincubation Number of revertants per plate (mean of 3 plates)

	TA1535			TA1537
Conc. µg/plate	— MA	+ MA	Cytotoxic (yes/no)	— MA	+  MA	Cytotoxic (yes/no)
0*	13.7	13	No	3	4.3	No
10	13.3	13.7	No	3	4.7	No
31.6	14.3	12	No	3	3.3	No
100	12.3	12.3	No	2.3	4.3	No
316	14.3	12	Yes	4	4.3	Yes
1000	0	12.7	Yes	0	0	Yes
Positive control	487.3	499.7	No	502	503	No

*solvent control with Ethylene glycol dimethylether

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with and without metabolic activation

[2-(Perfluorohexyl)ethyl]dichloro(methyl)silane has been tested in a reliable study, conducted according to OECD 471 and in compliance with GLP. No test-substance related increase in the number of revertants was observed with and without metabolic activation when tested up to cytotoxic concentration in Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102. The result of the initial plate incorporation assay was confirmed in an independent experiment using the pre-incubation method. Appropriate positive and solvent controls were included and gave expected results. It is concluded that the substance is negative for mutagenicity to bacteria under the conditions of the test.