[2-[[2-cyano-3-[4-[ethylbenzylamino]phenyl]-1-oxoallyl]oxy]ethyl]trimethylammonium chloride

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The substance was tested in an Ames test in Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 102 with and without metabolic activation. The maximum concentration tested was limited by a reduction of bacterial background lawn. In two independent experiments no biologically relevant increase in revertant colony numbers of any of the five tester strains was observed. Therefore the substance is considered non-mutagenic (Eurofins 2017).

In 1995 a very pure batch of the substance was tested in an Ames test with TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 with and without metabolic activation in a plate incorporation assay. In TA100 with metabolic activation a dose related > 2 -fold increase of the number of revertants was reported at non-cytotoxic concentrations. In all other strains and TA100 without metabolic activation no increase was reported (JBC 1995).

An additional Ames test with TA 1535, TA 1537, TA 98 and TA 100 with metabolic activation in a plate incorporation assay was negative in all strains tested (Bayer 1980)

In a HPRT assay according to OECD 476 the substance did not induce mutations in V79 cells both in presence and absence of metabolic activation. The substance was tested up to cytotoxic concentrations.

It can be concluded that the substance is negative in this test and does not induce mutations in mammalian cells (Bayer 2002).

In a chromosome aberration test V79 cells were exposed to the substance for 4 hours (with and without metabolic activation) or 21 hours (without metabolic activation). Three or four concentrations (maximum concentration based on 55± 5% cytotoxicity) per experiment were evaluated for the presence of chromosom aberrations. No increase in the number of aberrations compared to control and/or historical controls was observed. Positive controls were within historical ranges. The substance is considered not clastogenic in this assay (Eurofins 2017).

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

31 January 2002 to 27 April 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

July 21, 1997

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5300 - In vitro Mammalian Cell Gene Mutation Test

GLP compliance:

yes

Type of assay:

other: HPRT forward mutation in mammalian cells

Specific details on test material used for the study:

purified laboratory sample

Target gene:

6-thioguanine

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

CELLS USED
- Source of cells: University of Ulm, Germany
- Cell cycle length, doubling time or proliferation index: 10-14 hours
- Methods for maintenance in cell culture: stock kept under liquid nitrogen
- Modal number of chromosomes: 22


MEDIA USED
- Type and identity of media including CO2 concentration if applicable: maintained in plastic vessels at 37 °C and 5%CO2, recloned by subculturing twice weekly (checked regularly for karyotype and mycoplasma presence)
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability:yes

Metabolic activation:

with and without

Metabolic activation system:

S9 derived from Aroclor 1254 induced rat liver homogenate (concentration in test medium 5%)

Test concentrations with justification for top dose:

exp 1 without metabolic activation: 10, 20, 40, 60, 80 and 100 µg/mL
exp 2 without metabolic activation: 15, 30, 60, 90, 120 and 240 µg/mL
exp 1 with metabolic activation: 60, 80, 100, 120, 160, 200 and 240 µg/mL
exp 2 with metabolic activation: 60, 80, 100, 120, 160, 200 and 240 µg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: deionized water
- Justification for choice of solvent/vehicle: substance soluble up to 250 mg/mL in pre-test (precipitate in culture medium at 240 µg/mL and above)

Untreated negative controls:

yes

Remarks:

untreated

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

other: dimethylbenzanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: Exponentially growing cells were plated in culture medium in 250 mL flasks for 16-24 hours to allow attachment. Thereafter the cells were exposed for 5 hours. Monolayers were washed with PBS, trypsinized and replated in 20 mL culture medium (1.5E06 cells/250 mL) in flasks and in 3 petri dishes (5 mL medium with 200 cells/dish).
Petri dishes were incubated for 6 days and used to assess cytotoxicity (survival to treatment).
The cells in the flasks were incubated to permit growth and expression of mutations. After 6 days the cells were re-seeded in 8 petri dishes per culture at 3E05 cells/dish in presence of 6-thioguaninine. Three additional petri dishes with 200 cels/dish were used to calculate cloning efficiency.
After 6-8 days of incubation colonies were fixed, stained with Chiemsa and counted.

NUMBER OF REPLICATIONS: 8/concentration

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency and relative total growth

Evaluation criteria:

1) assessment in at least 5 replicates per concentration with relative survival, relative population growth and absolute cloning efficiency > 10%
2) positive result with dose related increase in mutant frequency (at least 2-3 times of vehicle controls) and this finding can be reproduced in a separate experiment
3) no unphysiological culture conditions (effects on pH and osmolality)
4) scientific judgement

Statistics:

ANOVA and Dunnet test

Key result

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

maximum concentrations based on cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: none
- Effects of osmolality: none
- Evaporation from medium: NA
- Precipitation: at and above 240 µg/mL

RANGE-FINDING/SCREENING STUDIES:without metabolic activation: 100% cell death at 100 µg/mL; with metabolic activation 100% cell death at 240 µg/mL

HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%)
- Positive historical control data: included in the report
- Negative (solvent/vehicle) historical control data: included in the report

ADDITIONAL INFORMATION ON CYTOTOXICITY:
- cloning efficiency, rel growth: no effects up to 240 µg/mL:
- survival to treatment: in all experiments with and without metabolic activation cell death (100%) at 240 µg/mL

Conclusions:

The substance did not induce mutations in mammalian cells

Executive summary:

In a HPRT assay according to OECD 476 the substance did not induce mutations in V79 cells both in presence and absence of metabolic activation. The substance was tested up to cytotoxic concentrations.

It can be concluded that the substance is negative in this test and does not induce mutations in mammalian cells.

Reference 2

Endpoint:

in vitro gene mutation study in bacteria

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 471 (Bacterial Reverse Mutation Assay)

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Target gene:

Histidine

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

CELLS USED
- Source of cells: TA 98, TA 1535 and TA 102 MOLTOX, INC., NC 28607, USA; TA 100 and TA 1537 Xenometrix AG, Switzerland.
- Methods for maintenance in cell culture if applicable: stock cultures stored with nutrient broth (OXOID) supplemented with DMSO (approx. 8% v/v) over liquid nitrogen

Metabolic activation:

with and without

Metabolic activation system:

liver microsomal fraction from rats treated with phenobarbital / β-naphthoflavone

Test concentrations with justification for top dose:

exp 1: 0.00316, 0.0100, 0.0316, 0.100, 0.316, 1.0 and 2.5 µL/plate (TA 98 and TA 100)
in addition for TA 1535, TA 1537 and TA 102: 0.00100 µL/plate
exp 2: 0.000316, 0.00100, 0.00316, 0.0100, 0.0316, 0.100, 0.316 and 1.0 µL/plate

Top doses limited by reduction of bacterial background lawn (without S9 at and above 0.1-1 ug/plate; with S9 at and above 0.1-2.5 ug/plate).

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: destilled water
- Justification for choice of solvent/vehicle: solubility

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

methylmethanesulfonate

other: 4-nitro-o-phenylene-diamine; 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: exp 1 plate incorporation; exp 2 preincubation

DURATION
- Preincubation period: 60 min at at 37 °C
- Incubation time: at 37 °C for at least 48 h in the dark

NUMBER OF REPLICATIONS: 3/concentration

DETERMINATION OF CYTOTOXICITY
- Method: reduction of bacterial background lawn

Evaluation criteria:

A test item is considered as mutagenic if:
- a clear and dose-related increase in the number of revertants occurs and/or
- a biologically relevant positive response for at least one of the dose groups occurs
in at least one tester strain with or without metabolic activation.
A biologically relevant increase is described as follows:
- if in tester strains TA 98, TA 100 and TA 102 the number of reversions is at least twice as high
- if in tester strains TA 1535 and TA 1537 the number of reversions is at least three times higher
than the reversion rate of the solvent control

Statistics:

NA

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 0.316 - 2.5 µL/plate

Vehicle controls validity:

valid

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 0.1-1.0 ug/plate

Vehicle controls validity:

valid

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 0.1-1.0 ug/plate

Vehicle controls validity:

valid

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 0.1-1.0 ug/plate

Vehicle controls validity:

valid

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 0.316-1.0 ug/plate

Vehicle controls validity:

valid

Positive controls validity:

valid

Conclusions:

The substance is considered non-mutagenic under the conditions of the test.

Executive summary:

The substance was tested in an Ames test in Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 102 with and without metabolic activation. The maximum concentration tested was limited by a reduction of bacterial background lawn. In two independent experiments no biologically relevant increase in revertant colony numbers of any of the five tester strains was observed. Therefore the substance is considered non-mutagenic.

Reference 3

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03 February 2017 to 24 April 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Type of assay:

other: in vitro chromosome aberration test in mammalian cells

Target gene:

NA

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

CELLS USED
- Source of cells: Eurofins Munich (ATCC, CCL-93)
- Cell cycle length: 12-14 hours
- Methods for maintenance in cell culture: stored under liquid nitrogen if applicable:
- Modal number of chromosomes: 2n=22

MEDIA USED
- Type and identity of media including CO2 concentration: MEM (minimum essential medium) supplemented with 10% FBS (fetal bovine serum) (5% CO2)
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes

Additional strain / cell type characteristics:

not applicable

Cytokinesis block (if used):

colcemid 17.5 hours after treatment

Metabolic activation:

with and without

Metabolic activation system:

The S9 liver microsomal fraction was prepared from phenobarbital/β-naphthoflavone induced rat liver homogenate

Test concentrations with justification for top dose:

exp 1 (4 h without metabolic activation): 25, 50, 75, 100, 125, 150, 175, 200 and 250 µg/mL (evaluated 25, 50, 75 and 100 µg/mL)
exp 1 (4 h with metabolic activation): 200, 250, 300, 325, 350, 375, 400, 425, 450 and 500(P) µg/mL (evaluated 250, 325 and 375 µg/mL)
exp 2 (21 h without metabolic activation): 25, 50, 75, 100, 125, 150, 175, 200 and 250 µg/mL (evaluated 50, 125 and 250 µg/mL

Vehicle / solvent:

Exp 1: MEM (minimum essential medium)
Exp 2: MEM (minimum essential medium) supplemented with 10% FBS

pH was adapted to 7 ± 0.4 with sodium hydroxide solution. For osmolality 354 mOsmol/kg was measured at 5 mg/mL

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

cyclophosphamide

ethylmethanesulphonate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in suspension
- Cell density at seeding: 1E04 cells/mL

DURATION
- Exposure duration: 4 hours (with and without metabolic activation) and 21 hours (without metabolic activation)
- Expression time (cells in growth medium): 17 h (4 hours experiments)
- Selection time (if incubation with a selection agent): colcemid 17.5 hours after start of treatment
- Fixation time (start of exposure up to fixation or harvest of cells): 21 hours after start of treatment

SPINDLE INHIBITOR (cytogenetic assays): colcemid

NUMBER OF REPLICATIONS: 2/concentration

PREPARATION: After ca 20 hours preparation was started. At first cells were trypsinated and resuspended in about 9 mL complete culture medium. An aliquot of each culture was removed to determine the cell count by a cell counter (AL-Systems).Then cultures were transferred into tubes and incubated with hypotonic solution (0.4% KCl) for 15-20 min. After hypotonic treatment the cells were fixed at least two times with 3 + 1 methanol + glacial acetic acid and spread onto the slides. After the fixation steps the slides were dried and stained with Giemsa. The slides were coverslipped using 2-3 drops of Eukitt(R). Afterwards they were air dried. 300 well-spread metaphases were microscopically analysed for cytogenetic damage

DETERMINATION OF CYTOTOXICITY
- Method: releative increase in cell count (RICC)

OTHER EXAMINATIONS:
- Determination of polyploidy: included

Rationale for test conditions:

based on pre-test at 5, 10, 25, 50, 100, 250, 500, 1000, 2500 and 5000 µg/mL (4 h with and without metabolic activation)

Evaluation criteria:

positive:
a) at least one of the test concentrations exhibits a statistically significant increase compared with the concurrent negative control,
b) the increase is dose-related when evaluated with an appropriate trend test,
c) any of the results are outside the distribution of the historical negative control data

negative:
a) none of the test concentrations exhibits a statistically significant increase compared with the concurrent negative control,
b) there is no concentration-related increase when evaluated with an appropriate trend test,
c) all results are inside the distribution of the historical negative control data.

Statistics:

Fisher´s exact test; Chi-square Test

Key result

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at and above 100 µg/mL (4 hours without metabolic activation) at and above 400 µg/mL (4 hours with metabolic activation) at and above 250 µg/mL (21 hours without metabolic activation)

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS: none

HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%): available in the report

ADDITIONAL INFORMATION ON CYTOTOXICITY:
- Measurement of cytotoxicity used: RICC

RICC compared to controls :
exp 1 without metabolic activation: 59% at 75 µg/mL, 34% at 100 µg/mL, 11% at 125 µg/mL and 0% and below at 150 µg/mL and higher
exp 1 with metabolic activation: 66% at 325 µg/mL, 49% at 350 µg/mL, 43% at 375 µg/mL, 22% at 400 µg/mL, 7% at 425 µg/mL and 0% and below at 450 µg/mL and higher
exp 2: without metabolic activation: 69% at 75 µg/mL, 54% at 100 µg/mL, 62% at 125 µg/mL, 56% at 150 µg/mL, 48% at 175 µg/mL, 50% at 200 µg/mL and 41% at 250 µg/mL

   

Experiment I,without metabolic activation

Negative Control versus Test Group	Concentration [µg/mL]	Treatment Time [h]	Aberrant Cells (excl. gap)	Significance	p Value
1	25	4	11	-	0.6422
2	50	4	9	-	1.0000
3	75	4	11	-	0.6422
4	100	4	7	-	1.0000
EMS	900	4	20	+	0.0315

Experiment I,with metabolic activation

Negative Control versus Test Group	Concentration [µg/mL]	Treatment Time [h]	Aberrant Cells (excl. gap)	Significance	p Value
2	250	4	11	-	0.8208
4	325	4	2	-	0.0630
6	375	4	9	-	1.0000
CPA	1.11	4	21	+	0.0377

 

Experiment II,without metabolic activation

NegativeControl versus Test Group	Concentration [µg/mL]	Treatment Time [h]	Aberrant Cells (excl. gap)	Significance	p Value
2	50	21	8	-	0.2222
5	125	21	10	-	0.0888
9	250	21	8	-	0.2222
EMS	400	21	24	+	<0.0001

 

 

 

 

 

Conclusions:

The substance did not induce aberrations in V79 cells. The substance is considered not clastogenic in this assay.

Executive summary:

In a chromosome aberration test V79 cells were exposed to the substance for 4 hours (with and without metabolic activation) or 21 hours (without metabolic activation). Three or four concentrations (maximum concentration based on 55 ± 5% cytotoxicity) per experiment were evaluated for the presence of chromosome aberrations. No increase in the number of aberrations compared to control and/or historical controls was observed. Positive controls were within historical ranges. The substance is considered not clastogenic in this assay.

Reference 4

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

August 24 1995 to September 7 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

plated in duplicate

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Version / remarks:

JMIT

Principles of method if other than guideline:

The use of duplicate plating was not justified

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Target gene:

histidine (Salmonella typhimurium) or tryptophan (E coli)

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

CELLS USED
- Source of cells: slamonella typhimurium from Bruce Ames, California University, E coli from Institute of medical Science, University of Tokyo

Metabolic activation:

with and without

Metabolic activation system:

S9 prepared from liver homogenate of rats treated with phenobarbital/5,6-benzoflavone

Test concentrations with justification for top dose:

exp 1 : 10, 50, 100, 500, 1000 and 5000 ug/plate (S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 with and without S9)
exp 2: 5, 10, 20, 39, 78 and 156 ug/plate (TA 1535 and TA 100 without S9); 10, 20, 39, 78, 156 and 313 ug/plate ( TA 1537 and TA 98 without S9); 39, 78, 156, 313, 625 and 1250 ug/plate (E. Coli without S9) ;
20, 39, 78, 156, 313 and 625 ug/plate (TA 1535 and TA 100 with S9); 39, 78, 156, 313, 625 and 1250 ug/plate ( TA 1537 and TA 98 with S9); 156, 313, 625, 1250, 2500 and 5000 ug/plate (E. Coli with S9)
exp 3: 5, 10, 20, 39, 78 and 156 ug/plate (TA 1535 and TA 100 without S9); 20, 39, 78, 156 313 and 625 ug/plate ( TA 1537 and TA 98 without S9); 20, 39, 78, 156 313 and 625 ug/plate (TA 1535 and TA 100 with S9):

Vehicle / solvent:

DMSO
- Justification for choice of solvent/vehicle: stability of substance in vehicle

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

benzo(a)pyrene

furylfuramide

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Preincubation period: NA
- Exposure duration:48 h at 37 °C

NUMBER OF REPLICATIONS: 2/concentration

DETERMINATION OF CYTOTOXICITY
- Method: not indicated

Evaluation criteria:

A test item is considered as mutagenic if:
- a significant and dose-related increase in the number of revertants occurs and the number of revertant colonies is at least twice as high with sufficient reproducibility

Statistics:

NA

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 313 ug/plate and above

Vehicle controls validity:

valid

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 100 ug/plate and above

Vehicle controls validity:

valid

Positive controls validity:

valid

Key result

Species / strain:

other: TA 1535, TA 1537, TA 98 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

see doses selected

Vehicle controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: dose related > twofold increase

Conclusions:

Under the conditions of the test the substance is considered mutagenic

Executive summary:

The substance was tested in an Ames test with TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 with and without metabolic activation in a plate incorporation assay. In TA100 with metabolic activation a dose related > 2 -fold increase of the number of revertants was reported at non-cytotoxic concentrations. In all other strains and TA100 without metabolic activation no increase was reported.

Based on this effect the substance has to be considered mutagenic to bacteria.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The results of the two Ames tests (Eurofins 2018 and Bayer 1980) are conflicting with one of the tests (JBC 1995) showing a positive result in one strain of Salmonella, TA100 with metabolic activation. As two Ames tests are available with negative results in this strain of Salmonella and all other strains tested are without any effects in all 3 tests, it can be concluded in a weight of evidence approach that the substance is negative in the Ames test.

Justification for classification or non-classification

Based on the information available the substance does not need to be classified for mutagenicity according toRegulation (EC) No 1907/2006.