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

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03 August 2011 to 20 December 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Version / remarks:

(1997)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5375 - In vitro Mammalian Chromosome Aberration Test

Version / remarks:

(1998)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)

Version / remarks:

(2008)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

Reference substance 001

Cas Number:

675-62-7

Constituent 2

Reference substance name:

Dichloromethyl(3,3,3-trifluoropropyl)silane

EC Number:

211-623-4

EC Name:

Dichloromethyl(3,3,3-trifluoropropyl)silane

Cas Number:

675-62-7

Molecular formula:

C4H7Cl2F3Si

IUPAC Name:

dichloro(methyl)(3,3,3-trifluoropropyl)silane

Method

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

- Type and identity of media: MEM
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes

Metabolic activation:

with and without

Metabolic activation system:

Liver S9 of Wistar phenobarbital and ß-naphthoflavone-induced rat liver S9 mix

Test concentrations with justification for top dose:

Pre-experiment:
with and without metabolic activation: 0.020, 0.039, 0.078, 0.16, 0.31, 0.63, 1.25, 2.5, 5.0, and 10.0 mM

Experiment I:
without metabolic activation: 0.63, 1.25, and 2.5 mM
with metabolic activation: 1.25, 2.5, and 5.0 mM
Experiment II:
without metabolic activation: 0.16, 0.63, and 1.75 mM
with metabolic activation: 2.0, 4.0, and 6.0 mM

Vehicle / solvent:

-Vehicle (s)/solvent(s) used: cell culture medium (MEM)
-Justification for choice of solvent/vehicle: The test item was dissolved in medium. The solvent was compatible with the survival of the cells and the S9 activity.

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

Remarks:

without metabolic activation: 400 and 900 µg/ml

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

with metabolic activation: 0.83 µg/ml

Details on test system and experimental conditions:

TREATMENT TIME:
4 hours (Experiment I with and without metabolic activation, experiment II with metabolic activation)
20 hours (Experiment II without metabolic activation)

FIXATION INTERVAL: 20 hours (Experiment I and II with and without metabolic activation)
NUMBER OF REPLICATIONS: 2 independent experiments
NUMBER OF CELLS SEEDED: 1E4 - 5E4 cells
NUMBER OF CULTURES: two cultures per concentration
NUMBER OF CELLS SCORED: 200 cells per concentration (100 cells per culture), (except in experiment I at a concentration of 0.63 mM without metabolic activation: 250 cells and in experiment II at a concentration of 2 mM with metabolic activation: 300 cells).
DETERMINATION OF CYTOTOXICITY: Mitotic index, cell density

Evaluation criteria:

There are several criteria for determining a positive result:
- a clear and dose-related increase in the number of cells with aberrations,
- a biologically relevant response for at least one of the dose groups, which is higher than the laboratory negative control range (0.0% - 4.0% aberrant cells (with and without metabolic activation)).

Statistics:

According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the results is not regarded as necessary.

Results and discussion

Test results

Key result

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Results of chromosome analysis
without metabolic activation
		Cytotoxicity	Chromatid aberrations				Isochromatid aberrations				rel. Mitotic index (%)	rel. Cell density (%)	Poly- ploidy	mean % aberrant cells
	Scored cells		gaps	breaks	inter- changes	other	gaps	breaks	inter- changes	other				incl. Gaps	excl. Gaps
Experiment I
solvent control	200	-	2	1	0	0	0	0	0	0	100	100	1	1.5	0.5
0.16 mM	-	no	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	83	n.d.	n.d.	n.d.	n.d.
0.31 mM	-	no	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	98	n.d.	n.d.	n.d.	n.d.
0.63 mM	250	no	5	4	1	0	0	0	2	0	93	93	3	4.8	2.8
1.25 mM	200	no	3	1	0	0	0	0	0	0	82	97	2	2.0	0.5
2.5 mM	200	yes	7	3	0	0	1	0	0	0	40	71	0	4.0	1.5
5.0 mM	-	yes	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	18	n.d.	n.d.	n.d.	n.d.
EMS 900 µg/mL	200	-	5	13	8	1	0	0	0	3	61	74	1	12.5	11.5
Experiment II
solvent control	200	-	1	0	0	0	0	0	2	0	100	100	0	1.5	1.0
0.08 mM	-	no	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	80	n.d.	n.d.	n.d.	n.d.
0.16 mM	200	no	3	0	0	0	1	0	0	0	85	92	2	2.0	0.0
0.31 mM	200	yes	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	56	n.d.	n.d.	n.d.	n.d.
0.63 mM	200	yes	0	0	0	0	0	0	0	0	54	77	1	0.0	0.0
1.25 mM	-	yes	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	46	n.d.	n.d.	n.d.	n.d.
1.75 mM	200	yes	5	2	1	1	0	0	1	0	48	56	0	4.5	2.5
2.5 mM	-	yes	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.
5.0 mM	-	yes	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.
EMS 400 µg/mL	200	-	0	12	5	0	1	0	1	1	54	98	1	8.5	8.0
Results of chromosome analysis
with metabolic activation
		Cytotoxicity	Chromatid aberrations				Isochromatid aberrations				rel. Mitotic index (%)	rel. Cell density (%)	Poly- ploidy	mean % aberrant cells
	Scored cells		gaps	breaks	inter- changes	other	gaps	breaks	inter- changes	other				incl. Gaps	excl. Gaps
Experiment I
solvent control	200	-	8	5	0	0	0	0	0	0	100	100	2	5.5	2.5
0.63 mM	-	no	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	98	n.d.	n.d.	n.d.	n.d.
1.25 mM	200	no	4	1	0	0	0	0	0	0	77	72	2	2.5	0.5
2.5 mM	200	yes	1	1	0	1	0	0	0	0	62	93	0	1.5	1.0
5.0 mM	200	yes	3	1	0	0	0	0	0	0	40	76	2	2.0	0.5
7.5 mM	-	yes	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	14	n.d.	n.d.	n.d.	n.d.
10.0 mM	-	yes	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.
CPA 0.83 µg/mL	200	-	3	9	6	1	1	1	0	1	74	81	1	10.0	8.5
Experiment II
solvent control	200	-	2	1	2	1	0	0	0	0	100	100	2	2.5	1.5
0.5 mM	-	no	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	103	n.d.	n.d.	n.d.	n.d.
1.0 mM	-	no	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	80	n.d.	n.d.	n.d.	n.d.
2.0 mM	300	no	3	6	0	1	0	0	2	0	87	98	2.5	3.7	3.0
4.0 mM	200	no	3	2	0	1	1	0	2	0	87	99	3	4.0	2.5
6.0 mM	200	yes	7	3	0	1	1	0	1	0	46	80	2	6.5	2.5
8.0 mM	-	yes	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	10	n.d.	n.d.	n.d.	n.d.
10.0 mM	-	yes	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.	3	n.d.	n.d.	n.d.	n.d.
CPA 0.83 µg/mL	200	-	0	10	5	1	1	0	0	2	88	98	1	9.0	8.5

n.d. not determined

Applicant's summary and conclusion

Conclusions:

Dichloromethyl(3,3,3-trifluoropropyl)silane was tested for in vitro cytogenicity to mammalian cells according to the OECD TG 473 and in compliance with GLP. The test item did not induce structural chromosomal aberrations in the V79 Chinese hamster cell line with and without metabolic activation up to cytotoxic concentrations. Appropriate positive and solvent controls were included into the study and gave the expected results. The test item is therefore considered to be non-clastogenic under the conditions of the test.