Alkyl iodides, C8-14, γ-ω-perfluoro

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:

2010 -08-09 till 2010-08-24

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline-conform study under GLP without deviations

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)

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:

Automatically generated during migration to IUCLID 6, no data available

IUPAC Name:

Automatically generated during migration to IUCLID 6, no data available

Details on test material:

Identity: Fluowet El 812
CAS No. 85995-91-1
Batch No.: T 7362
Description: wax like solid
Colour: white to slight violet
Purity: 92.5 %
Dose calculations adjusted to purity
Stability in solvent: Not indicated by the sponsor
Storage: At room temperature, light protected
Expiration Date: October 28, 2011

Method

Species / strain

Species / strain / cell type:

other: TA 1535, TA 1537, TA 98, TA 100, WP2 uvrA

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Phenobarbital/ß-Naphthoflavone induced rat liver S9

Test concentrations with justification for top dose:

3, 10; 33; 100; 333; 1000; 2500; and 5000 µg/plate / pre-experiment/experiment I
10, 33; 100; 333; 1000; 2500; and 5000 µg/plate / experiment II

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Acetone
- Justification for choice of solvent/vehicle: recommended by the sponsor

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: sodium azide; 4-nitro-o-phenylene-diamine; methyl methane sulfonate, 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: plate incorporation; preincubation;


DURATION
- Preincubation period: 1 hour
- Exposure duration: 72 hours


NUMBER OF REPLICATIONS: 3 plates


DETERMINATION OF CYTOTOXICITY
A reduction in the number of spontaneous revertants (below the induction factor of 0.5) or a clearing of the bacterial background lawn.

Evaluation criteria:

A test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA 98, TA 100, and WP2 uvrA) or thrice (strains TA 1535 and TA 1537) the colony count of the corresponding solvent control is observed.
A dose dependent increase is considered biologically relevant if the threshold is exceeded at more than one concentration.
An increase exceeding the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment.
A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls such an increase is not considered biologically relevant.

Statistics:

According to the OECD guideline 471, a statistical analysis of the data is not mandatory.

Results and discussion

Test results

Species / strain:

other: TA 1535, TA 1537, TA 98, TA 100, WP2 uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation:
The test item precipitated in the overlay agar in the test tubes from 1000 µg/plate up to 5000 µg/plate in experiment I and from 2500 µg/plate up to 5000 µg/plate in experiment II. Precipitation on the incubated agar plates was observed from 1000 µg/plate up to 5000 µg/plate in both experiments with and without S9 mix. The undissolved particles had no influence on the data recording.
- Other confounding effects:
COMPARISON WITH HISTORICAL CONTROL DATA: performed
ADDITIONAL INFORMATION ON CYTOTOXICITY: no toxic effects observed

Remarks on result:

other: other: reverse mutation assay

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

1.1     Summary of Results Pre-Experiment and Experiment I

Study Name: 1358102	Study Code: Harlan CCR 1358102
Experiment: 1358102 VV Plate	Date Plated: 09/08/2010
Assay Conditions:	Date Counted: 12/08/2010 to 16/08/2010

Metabolic Activation	Test Group	Dose Level (per plate)		Revertant Colony Counts (Mean ±SD)
				TA 1535	TA 1537	TA 98	TA 100	WP2 uvrA
Without Activation	Acetone			13 ± 1	7 ± 1	27 ± 5	147 ± 6	55 ± 4
	Untreated			18 ± 3	10 ± 2	29 ± 5	133 ± 10	61 ± 11
	Fluowet El 812	3 µg		12 ± 5	8 ± 5	24 ± 5	138 ± 19	55 ± 1
		10 µg		16 ± 3	8 ± 2	29 ± 2	152 ± 8	63 ± 2
		33 µg		14 ± 5	8 ± 1	28 ± 1	137 ± 13	57 ± 6
		100 µg		13 ± 4	10 ± 3	30 ± 4	136 ± 4	56 ± 6
		333 µg		14 ± 2	9 ± 1	28 ± 4	137 ± 4	63 ± 10
		1000 µg		13 ± 3^P	7 ± 3^P	32 ± 3^P	136 ± 11^P	54 ± 3^P
		2500 µg		13 ± 2^P M	5 ± 2^P M	24 ± 1^P M	149 ± 17^P	60 ± 8^P
		5000 µg		11 ± 2^P M	5 ± 2^P M	15 ± 2^P M	128 ± 8^P M	42 ± 1^P M
	NaN3	10 µg		1721 ± 110			1935 ± 118
	4-NOPD	10 µg				301 ± 17
	4-NOPD	50 µg			67 ± 4
	MMS	3.0 µL						1170 ± 22
With Activation	Acetone			23 ± 3	14 ± 2	36 ± 9	164 ± 14	66 ± 10
	Untreated			20 ± 6	17 ± 2	39 ± 11	151 ± 23	78 ± 11
	Fluowet El 812	3 µg		21 ± 8	12 ± 6	40 ± 9	157 ± 6	70 ± 8
		10 µg		24 ± 3	12 ± 2	38 ± 3	141 ± 5	81 ± 8
		33 µg		19 ± 6	13 ± 2	40 ± 12	141 ± 12	73 ± 8
		100 µg		19 ± 4	13 ± 2	37 ± 7	151 ± 4	80 ± 1
		333 µg		26 ± 6	17 ± 2	48 ± 4	140 ± 9	81 ± 9
		1000 µg		24 ± 9^P	16 ± 1^P	35 ± 6^P M	158 ± 4^P	86 ± 9^P
		2500 µg		20 ± 4^P M	13 ± 3^P M	35 ± 6^P M	170 ± 3^P	70 ± 7^P M
		5000 µg		13 ± 3^P M	7 ± 1^P M	25 ± 7^P M	131 ± 16^P M	69 ± 9^P M
	2-AA	2.5 µg		358 ± 5	545 ± 41	2089 ± 430	3173 ± 59
	2-AA	10.0 µg						305 ± 48

Key to Positive Controls		Key to Plate Postfix Codes
NaN3 2-AA 4-NOPD MMS	sodium azide 2-aminoanthracene 4-nitro-o-phenylene-diamine methyl methane sulfonate	P M	Precipitate Manual count

1.2     Summary of Results Experiment II

Study Name: 1358102	Study Code: Harlan CCR 1358102
Experiment: 1358102 HV2 Pre	Date Plated: 18/08/2010
Assay Conditions:	Date Counted: 24/08/2010

Metabolic Activation	Test Group	Dose Level (per plate)		Revertant Colony Counts (Mean ±SD)
				TA 1535	TA 1537	TA 98	TA 100	WP2 uvrA
Without Activation	Acetone			13 ± 6	18 ± 5	32 ± 10	117 ± 16	45 ± 3
	Untreated			14 ± 2	16 ± 3	35 ± 3	130 ± 16	43 ± 9
	Fluowet El 812	10 µg		12 ± 3	16 ± 5	34 ± 3	129 ± 1	47 ± 1
		33 µg		13 ± 2	15 ± 2	32 ± 9	124 ± 8	43 ± 3
		100 µg		13 ± 2	16 ± 1	29 ± 4	120 ± 20	48 ± 1
		333 µg		12 ± 4	11 ± 4	35 ± 5	129 ± 19	39 ± 10
		1000 µg		17 ± 2^P	18 ± 3^P	37 ± 3^P	131 ± 7^P	41 ± 5^P
		2500 µg		9 ± 3^P M	16 ± 1^P	33 ± 3^P M	151 ± 5^P	37 ± 3^P M
		5000 µg		9 ± 3^P M	12 ± 2^P M	24 ± 5^P M	133 ± 7^P M	36 ± 3^P M
	NaN3	10 µg		1767 ± 37			2020 ± 98
	4-NOPD	10 µg				433 ± 26
	4-NOPD	50 µg			78 ± 5
	MMS	3.0 µL						498 ± 21
With Activation	Acetone			24 ± 7	20 ± 3	47 ± 7	133 ± 9	58 ± 4
	Untreated			26 ± 1	29 ± 6	43 ± 8	121 ± 5	56 ± 11
	Fluowet El 812	10 µg		26 ± 7	25 ± 4	47 ± 8	138 ± 7	54 ± 16
		33 µg		19 ± 5	24 ± 3	47 ± 2	141 ± 13	64 ± 8
		100 µg		24 ± 6	26 ± 3	40 ± 3	139 ± 33	56 ± 6
		333 µg		20 ± 4	25 ± 6	49 ± 9	118 ± 9	48 ± 7
		1000 µg		23 ± 2^P M	28 ± 2^P	55 ± 5^P	135 ± 16^P	53 ± 12^P
		2500 µg		16 ± 4^P M	17 ± 3^P M	36 ± 1^P M	133 ± 10^P M	63 ± 5^P
		5000 µg		15 ± 3^P M	13 ± 4^P M	35 ± 5^P M	122 ± 8^P M	46 ± 5^P M
	2-AA	2.5 µg		350 ± 11	271 ± 11	1799 ± 45	1809 ± 39
	2-AA	10.0 µg						395 ± 10

Key to Positive Controls		Key to Plate Postfix Codes
NaN3 2-AA 4-NOPD MMS	sodium azide 2-aminoanthracene 4-nitro-o-phenylene-diamine methyl methane sulfonate	P M	Precipitate Manual count

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with and without S9 mix

In conclusion, it can be stated that during the described mutage¬nicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.

Executive summary:

The test item Fluowet El 812 was assessed for its potential to induce gene mutations in the plate incorporation test (experi­ment I) and the pre-incubation test (experiment II) using Salmo­nella typhimurium strains TA 1535, TA 1537, TA 98, and TA 100, and the Escheri­chia coli strain WP2 uvrA.

The assay was performed in two independent experiments both with and without liver microsomal activation. Each concentration and the controls were tested in triplicate. The test item was tested at the following concentrations:

Pre-Experiment/Experiment I:           3; 10; 33; 100; 333; 1000; 2500; and 5000 µg/plate

Experiment II:                                   10, 33; 100; 333; 1000; 2500; and 5000 µg/plate

The plates incubated with the test item showed normal background growth up to 5000 µg/plate with and without S9 mix in both experiments.

No toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), occurred in the test groups with and without metabol­ic activation.

No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with Fluowet El 812 at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher muta­tion rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.

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