Carboxymethyldimethyl-3-[[(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)sulphonyl]amino]propylammonium hydroxide

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

In Vitro (Mutagenic effects - bacterial): OECD 471; Ames study. Negative. Reliability = 1.

In Vitro (Clastogenic effects - mammalian): OECD 473; Chromosome aberrations in CHL/IU. Negative. Reliability = 2

In Vitro (Mutagenic effects - mammalian): OECD 476; in vitro mammalian cell gene mutation test (mouse lymphoma assay); Negative. Reliability = 1.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Deviations:

no

Remarks:

The study was conducted according to the guideline in effect at the time of study conduct.

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Target gene:

L5178Y/TK+/- mouse lymphoma cells are heterozygous at the normally diploid thymidine kinase (TK) locus.

Species / strain / cell type:

mouse lymphoma L5178Y cells

Details on mammalian cell type (if applicable):

- Type and identity of media: cells were prepared in 50% conditioned F0P supplemented with 10% horse serum and 2 mM L-glutamine (F10P) and 50% Fischer's Media for Leukemic Cells of Mice with 0.1% Pluronics F-68 (F0P). All media contained antibiotics.
- Properly maintained: yes, Each batch of frozen cells was tested and found to be free of mycoplasma contamination.
- Periodically "cleansed" against high spontaneous background: yes, Prior to use in the assay, L5178Y/TK+/- cells were cleansed to reduce the frequency of spontaneously occurring TK-/- cells.

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver S9

Test concentrations with justification for top dose:

Preliminary Toxicity Assay: 0.977 to 500 μg/mL
Mutagenicity Assay: 0.977 to 500 μg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: The vehicle of choice, ethanol (EtOH), permitted preparation of a workable suspension at a concentration of 50 mg/mL.

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

7,12-dimethylbenzanthracene

methylmethanesulfonate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- For the PRELIMINARY TOXICITY ASSAY only, after a 4-hour treatment in the presence and absence of S9 activation, cells were washed with culture medium and cultured in suspension for two days post-treatment, with cell concentration adjustment on the first day. After a 24-hour treatment in the absence of S9 activation, cells were washed with culture medium and immediately readjusted to 3x10e5 cells/mL. Cells were then cultured in suspension for an additional two days post-treatment with cell concentration adjustment on the first day.
- For the DEFINITIVE ASSAY only, at the end of the exposure period, the cells were washed with culture medium and collected by centrifugation. The cells were resuspended in 20 mL F10P on Day 1 and in 10 mL F10P on Day 2, and incubated at 37 ± 1°C for two days following treatment. Cell population adjustments to 3x105 cells/mL were made as follows:
* 4 hour treatment – 1 and 2 days after treatment.
* 24 hour treatment – immediately after test substance removal, and 2 and 3 days after treatment.

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: The cytotoxic effects of each treatment condition were expressed relative to the vehicle-treated control for suspension growth over two days post-treatment and for total growth (suspension growth corrected for plating efficiency at the time of selection).

Evaluation criteria:

A result was considered positive if a concentration-related increase in mutant frequency was observed in the treated cultures and one or more treatment conditions with 10% or greater total growth exhibit induced mutant frequencies of ≥90 mutants per 10e6 clonable cells (based on the average mutant frequency of duplicate cultures). If the average vehicle control mutant frequency was >90 mutants per 10e6 clonable cells, a doubling of mutant frequency over the vehicle would also be required.

A result was considered negative if the treated cultures exhibit induced mutant frequencies of less than 90 mutants per 10e6 clonable cells (based on the average mutant frequency of duplicate cultures) and there was no concentration-related increase in mutant frequency.

Key result

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- the test substance did not have an adverse impact on the pH or osmolality of the cultures.

Based on the results of the preliminary toxicity assay, cultures were treated in the mutagenicity assay at concentrations of 31.3, 62.5, 125, 250, 375 and 500 μg/mL (4-hour treatments with and without S9), and 0.977, 1.95, 3.91, 7.81, 15.6, 31.3, 46.9, 62.5 and 93.8 μg/mL (24-hour treatment without S9). No visible precipitate was observed at the beginning or end of treatment. Cultures treated at concentrations of 62.5, 125, 250, 375 and 500 μg/mL (4-hour treatments with and without S9), and 15.6, 31.3, 46.9 and 62.5 μg/mL (24-hour treatment without S9) exhibited 64 to 94%, 37 to 96%, and 19 to 79% RSG (relative suspension growth), respectively, and were cloned (cultures treated at other lower concentrations were discarded prior to cloning because a sufficient number of higher concentrations was available; cultures treated at other higher concentrations were excluded from evaluation of mutagenicity due to excessive toxicity). Relative total growth of the cloned cultures ranged from 66 to 107% (4-hour treatment with S9), 33 to 72% (4-hour treatment without S9) and 15 to 58% (24-hour treatment without S9). No increases in induced mutant frequency ≥90 mutants per 10e6 clonable cells were observed under any treatment condition.

Conclusions:

The results indicate that the test substance was negative in the L5178Y/TK+/- Mouse Lymphoma Assay under the conditions, and according to the criteria, of the test protocol.

This study and the conclusions which are drawn from it fulfil the quality criteria (validity, reliability, repeatability).

Executive summary:

The test substance was evaluated to determine its ability to induce forward mutations at the thymidine kinase locus in L5178Y mouse lymphoma cells in the presence and absence of Aroclor-induced rat liver S9 in accordance with OECD guideline 476. The test substance was prepared in ethanol and evaluated in a preliminary toxicity assay at concentrations from 1.95 to 500 μg/mL using 4-hour treatments with and without S9, and at concentrations from 0.977 to 250 μg/mL using a 24-hour treatment without S9. Based on the results, cultures were treated in the mutagenicity assay at concentrations of 31.3, 62.5, 125, 250, 375 and 500 μg/mL (4-hour treatments with and without S9), and 0.977, 1.95, 3.91, 7.81, 15.6, 31.3, 46.9, 62.5 and 93.8 μg/mL (24-hour treatment without S9). No visible precipitate was observed at the beginning or end of treatment.

No increases in induced mutant frequency ≥90 mutants per 10e6 clonable cells were observed under any treatment condition. All positive and vehicle control values were within acceptable ranges, and all criteria for a valid study were met. These results indicate the test substance was negative in the L5178Y/TK+/- Mouse Lymphoma Assay, under the conditions, and according to the criteria, of the test protocol.

Reference 2

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

Without activation: 0, 9.77, 19.5, 39.1, 78.1, 156, 313, 625, 1250 µg/plate
With activation: 0, 39.1, 78.1, 156, 313, 625, 1250, 2500, 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Ethanol soluble; DMSO (50 mg/mL suspension showed no discoloration, heat emission, etc., up to 2 h after preparation, thus judged stable

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

other: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide; 2-methoxy-6-chloro-9-[3-(2-chloroethyl)aminopropylamino]acridine2HCl; 2-aminoanthracene

Details on test system and experimental conditions:

Testing method: Preincubation method
Testing Conditions:
Composition: 0.1 mL bacterial suspension; 0.1 mL test substance solution; 0.5 mL Na phosphate buffer solution; 0.5 mL S9 mix (in metabolism activation method); 2.0 mL top agar
Preincubation: 20 minutes at 37±0.5°C
Incubation: 48 hours at 37±0.5°C
Colony Counting Method: Manual and instrumental with no correction

Evaluation criteria:

A substance was judged to be negative if the number of back-mutation colonies was less than 2 times the negative control.

Statistics:

None

Key result

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

≥625 µg/plate TA100, TA1535 and TA1537 without activation; ≥2500 µg/plate TA100, TA1535 and TA1537 with activation

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results (Main Test)
Metabolism Activation		Test substance dosage (µg/plate)	Back mutation (No. of colonies/plate)
			Base pair substitution						Frame shift
-S9			TA 100		TA1535		WP2uvrA		TA98		TA1537
		Negative control	107 107	115 (110)	11 11	9 (10)	28 28	29 (28)	21 19	27 (22)	7 7		5 (6)
		9.77	111 103	(107)	13 8	(11)	−		−		7 3		(5)
		19.5	114 119	(117)	7 8	(8)	−		−		8 6		(7)
		39.1	110 127	(119)	7 6	(7)	27 23	(25)	16 19	(18)	10 8		(9)
		78.1	110 98	(104)	9 8	(9)	30 26	(28)	18 19	(19)	7 9		(8)
		156	107* 73*	(90)	7 4	(6)	23 24	(24)	16 23	(20)	5 6		(6)
		313	102* 104*	(103)	6* 3*	(5)	24 23	(24)	10 17	(14)	6* 2*		(4)
		625	−		−		18 21	(20)	23 21	(22)	−
		1250	−		−		30* 22*	(26)	13* 14*	(14)	−
		Negative control	119 118	114 (117)	9 11	9 (10)	29 33	29 (30)	41 46	45 (44)	13 15	19 (16)
+S9		39.1	106 126	(116)	12 6	(9)	−		−		17 18	(18)
		78.1	124 117	(121)	11 3	(7)	−		−		26 19	(23)
		156	123 133	(128)	12 6	(9)	32 29	(31)	36 39	(38)	16 14	(15)
		313	104 129	(117)	9 9	(9)	35 27	(31)	35 27	(31)	22 24	(23)
		625	90 108	(99)	15 9	(12)	36 33	(35)	43 35	(39)	25 16	(21)
		1250	95* 100*	(98)	8* 7*	(8)	33 27	(30)	32 39	(36)	19* 21*	(20)
		2500	−		−		35 40	(38)	31 42	(37)	−
		5000	−		−		34* 33*	(34)	5* 15*	(10)	−
Positive control	-S9	Name	AF-2		NaN3		AF-2		AF-2		ICR-191
		Dosage (µg/plate)	0.01		0.5		0.01		0.1		0.5
		Colonies/plate	300 287	(294)	275 280	(278)	162 200	(181)	613 572	(593)	825 696	(761)
	+S9	Name	2AA		2AA		2AA		2AA		2AA
		Dosage (µg/plate)	1		2		10		0.5		2
		Colonies/plate	861 682	(772)	175 177	(176)	534 509	(522)	481 493	(487)	263 293	(278)
( ): average value of number of colonies of each plate *: growth inhibition of bacteria was observed AF-2: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide NaN3: sodium azide ICR-191: 2-methoxy-6-chloro-9-[3-(2-chloroethyl)aminopropylamino]acridine · 2HCl 2AA: 2-aminoanthracene

Conclusions:

The test substance was negative for mutagenicity.
This study and the conclusions which are drawn from it fulfil the quality criteria (validity, reliability, repeatability).

Executive summary:

The mutagenic potential of the test substance was examined. The test substance was exposed to concentrations of 0, 9.77, 19.5, 39.1, 78.1, 156, 313, 625, and 1250 µg/plate without S9 activation and 0, 39.1, 78.1, 156, 313, 625, 1250, 2500, and 5000 µg/plate with S9 activation. The test substance was negative for mutagenicity.

Reference 3

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Species / strain / cell type:

other: CHL/IU

Details on mammalian cell type (if applicable):

- Type and identity of media: Eagle MEM
- Properly maintained: yes

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

Short-term treatment : 0, 500, 1580, 5000 µg/mL
Continuous treatment: 0, 19.5, 39.1, 78.1 µg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Distilled water
- Justification for choice of solvent/vehicle: 50 mg/mL in distilled water showed a good suspension state; furthermore, there was no heat emission or discoloration at room temperature up to 2 hours after preparation

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

cyclophosphamide

mitomycin C

Details on test system and experimental conditions:

Short-term treatment test:
Incubator: 60 mm round plastic dish; 3 mL culture solution/dish (recovery period after treatment was 5 mL/dish); 2 incubators/dose were used
Cells: 1.5E4 cells/mL; incubation duration was 2 days
Treatment conditions: 0.3 mL test substance solution/dish; 0.5 mL S9/dish (for test with activation); treatment time was 6 hours; recovery time was 18 hours
Cell growth measurement: Calculated from the number of cells counted by electric cell counter.

Continuous treatment test:
Incubator: 60 mm round plastic dish; 5 mL culture solution/dish; 2 incubators/dose were used
Cells: 1.5E4 cells/mL; incubation duration was 2 days
Cell growth inhibition measurement: Electric cell counter was used for counting the number of cells
Treatment conditions: 0.5 mL test substance solution/dish; treatment time was 24 hours; recovery time was 0 hours
Cell growth inhibition measurement: Number of cells counted by electric cell counter

Evaluation criteria:

A positive judgment is given when the appearance frequency of cells having structural aberrations and numerical aberrations increased by more than 10% with a dependence on the dosage or when an increase exceeding 5% was regenerated in the chromosome aberration test and confirmation test; all others are judged as negative.

Key result

Species / strain:

other: CHL/IU

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

Additional information on results:

ADDITIONAL INFORMATION ON CYTOTOXICITY:
Cell growth in the short-term treatment without activation was 100, 93.3, 84.7, 84.2, 70.9, 66.9, 59.1, 53.0, 64.2, and 59.7 at 0, 19.5, 39.1, 78.1, 156, 313, 625, 1250, 2500, and 5000 µg/mL, respectively.

Cell growth in the short-term treatment with activation was 100, 101.9, 90.5, 92.3, 86.4, 87.4, 79.1, 75.3, 72.6, and 66.0 at 0, 19.5, 39.1, 78.1, 156, 313, 625, 1250, 2500, and 5000 µg/mL, respectively.

Cell growth in the continuous treatment without activation was 100, 95.2, 98.8, 91.5, 83.2, 61.7, 33.5, 24.0, 16.5, and 11.7 at 0, 19.5, 39.1, 78.1, 156, 313, 625, 1250, 2500, and 5000 µg/mL, respectively.

Test substance precipitation occurred at the beginning and end of the treatment at doses ≥1250 µg/mL at all treatment conditions.

Chromosome aberration test results (short-term treatment)
Treatment time (S9 mix)	Dosage (µg/mL)	Number of cells with chromosome structural aberrations (appearance frequency %)							Gap appearance frequency (appearance frequency %)	Cell growth (%)	Number of cells of [with] chromosome numerical aberrations (appearance frequency %)
		Number of cells observed	Chromosome fragment cleavage	Chromosome fragment exchange	Chromosome cleavage	Chromosome exchange	Other	Total number of aberration cells			Number of cells observed	Multiple	Other	Total number of aberration cells
6-18 (−)	Negative control (D.W.) 0	100	0	0	1	0	0	1	0	100	100	1	0	1
		100	2	0	0	0	0	2	1		100	2	0	2
		200	2 (1.0)	0 (0.0)	1 (0.5)	0 (0.0)	0 (0.0)	3 (1.5)	1 (0.5)		200	3 (1.5)	0 (0.0)	3 (1.5)
6-18 (−)	500	100	1	0	0	0	0	1	0	79.9	100	0	0	0
		100	1	1	0	0	0	2	0	76.5	100	1	0	1
		200	2 (1.0)	1 (0.5)	0 (0.0)	0 (0.0)	0 (0.0)	3 (1.5)	0 (0.0)	(78.2)	200	1 (0.5)	0 (0.0)	1 (0.5)
6-18 (−)	1580	100	0	0	1	0	0	1	1	72.6	100	0	0	0
		100	3	0	0	0	0	3	0	75.0	100	3	0	3
		200	3 (1.5)	0 (0.0)	1 (0.5)	0 (0.0)	0 (0.0)	4 (2.0)	1 (0.5)	(73.8)	200	3 (1.5)	0 (0.0)	3 (1.5)
6-18 (−)	5000*	100	1	0	0	0	0	1	1	77.8	100	2	0	2
		100	1	0	0	0	0	1	1	68.9	100	0	0	0
		200	2 (1.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	2 (1.0)	2 (1.0)	(73.4)	200	2 (1.0)	0 (0.0)	2 (1.0)
6-18 (−)	Positive control (MMC) 0.1	100	31	40	1	0	0	57	3	−	100	1	0	1
		100	19	37	3	0	0	52	1		100	0	0	0
		200	50 (25.0)	77 (38.5)	4 (2.0)	0 (0.0)	0 (0.0)	109 (54.5)	4 (2.0)		200	1 (0.5)	0 (0.0)	1 (0.5)
6-18 (+)	Negative control (D.W.) 0	100	1	1	0	0	0	2	1	100	100	0	0	0
		100	0	0	0	0	0	0	0		100	1	0	1
		200	1 (0.5)	1 (0.5)	0 (0.0)	0 (0.0)	0 (0.0)	2 (1.0)	1 (0.5)		200	1 (0.5)	0 (0.0)	1 (0.5)
6-18 (+)	500	100	0	0	0	0	0	0	1	90.4	100	0	0	0
		100	0	0	0	0	0	0	0	92.8	100	1	0	1
		200	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	1 (0.5)	(91.6)	200	1 (0.5)	0 (0.0)	1 (0.5)
6-18 (+)	1580	100	0	0	1	0	0	1	0	91.8	100	1	0	1
		100	0	0	0	0	0	0	0	90.4	100	2	0	2
		200	0 (0.0)	0 (0.0)	1 (0.5)	0 (0.0)	0 (0.0)	1 (0.5)	0 (0.0)	(91.1)	200	3 (1.5)	0 (0.0)	3 (1.5)
6-18 (+)	5000*	100	1	0	0	0	0	1	0	71.7	100	0	0	0
		100	0	1	0	0	0	1	0	79.2	100	0	0	0
		200	1 (0.50	1 (0.5)	0 (0.0)	0 (0.0)	0 (0.0)	2 (1.0)	0 (0.0)	(75.5)	200	0 (0.0)	0 (0.0)	0 (0.0)
6-18 (+)	Positive control (CPA) 6	100	32	42	3	0	0	54	1	−	100	1	0	1
		100	39	64	0	0	0	75	1		100	0	0	0
		200	71 (35.5)	106 (53.0)	3 (1.5)	0 (0.0)	0 (0.0)	129 (64.5)	2 (1.0)		200	1 (0.5)	0 (0.0)	1 (0.5)
The treatment duration is treatment time – recovery time. Data for aberration cell numbers in plates of each group are given in the first and second rows, with their sum in the third row. The numerical value of cell growth in plates of each test substance group are given in the first and second rows, with their sum in the third row. D. W.: distilled water MMC: mitomycin C CPA: dichlorophosphamide hydrate * Test substance precipitation occurred at the beginning and end of the treatment.

Conclusions:

The test substance was negative for chromosome aberrations.
This study and the conclusions which are drawn from it fulfil the quality criteria (validity, reliability, repeatability).

Executive summary:

The test substance was examined for its potential to induce chromosomal aberrations. The test substance was exposed to concentrations of 0, 500, 1580, and 5000 µg/mL in the short-term treatment and 0, 19.5, 39.1, and 78.1 µg/mL in the continuous treatment. The test substance was negative for the induction of chromosome aberrations.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

The mutagenic potential of the test substance was examined in bacterial cells. S. typhimurium TA1535, TA1537, TA98, TA100, and E. coli WP2 were exposed to the test substance at concentrations of 0, 9.77, 19.5, 39.1, 78.1, 156, 313, 625, and 1250 µg/plate without S9 activation and 0, 39.1, 78.1, 156, 313, 625, 1250, 2500, and 5000 µg/plate with S9 activation. No positive mutagenic responses were observed at any dose level or with any tester strain in either the absence or presence of S9 metabolic activation. The test substance was negative for mutagenicity.

The test substance was examined for its potential to induce chromosomal aberrations in vitro. CHL/IU cells were exposed to the test substance at concentrations of 0, 500, 1580, and 5000 µg/mL in the short-term treatment and 0, 19.5, 39.1, and 78.1 µg/mL in the continuous treatment. No increase in the number of structural or numerical aberrations was observed. The test substance was negative for the induction of chromosome aberrations.

 

The test substance was evaluated to determine its ability to induce forward mutations at the thymidine kinase locus in L5178Y mouse lymphoma cells in the presence and absence of Aroclor-induced rat liver S9. The test substance was prepared in ethanol and evaluated in a preliminary toxicity assay at concentrations from 1.95 to 500 μg/mL using 4-hour treatments with and without S9, and at concentrations from 0.977 to 250 μg/mL using a 24-hour treatment without S9. Based on the results, cultures were treated in the mutagenicity assay at concentrations of 31.3, 62.5, 125, 250, 375 and 500 μg/mL (4-hour treatments with and without S9), and 0.977, 1.95, 3.91, 7.81, 15.6, 31.3, 46.9, 62.5 and 93.8 μg/mL (24-hour treatment without S9). No visible precipitate was observed at the beginning or end of treatment. No increases in induced mutant frequency ≥90 mutants per 10E6 clonable cells were observed under any treatment condition. These results indicate the test substance was negative in the L5178Y/TK+/- Mouse Lymphoma Assay.

Justification for classification or non-classification

The test substance was negative for mutagenicity and clastogenicity in vitro in bacterial and mammalian cells. Therefore, the substance does not need to be classified for mutagenicity according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.