4,6-dimethyl-2-(1-phenylethyl)-3,6-dihydro-2H-pyran

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames test (OECD TG 471): negative

in vitro chromosome aberration test (OECD TG 473): negative

Link to relevant study records

Reference

Reference 1

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:

2015-04-17 till 2015-09-14

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:

other: US Food and Drug Administration, Center for Food Safety & Applied Nutrition, Redbook 2000 Toxicological Principles for the safety of Food Ingredients. IV.C.1.a. Bacterial Reverse Mutation Test, July 2000

GLP compliance:

yes (incl. QA statement)

Remarks:

TNO Triskelion, Utrechtseweg 48, 3704 HE Zeist, The Netherlands

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

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

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver homogenate (S9-mix)

Test concentrations with justification for top dose:

First bacterial reverse mutation test:
Experiment 1:
0, 21, 62, 185, 556, 1667, 5000 μg/plate (all strains with and without S9-mix)
Experiment 2:
0, 4.1, 12, 37, 111, 333, 1000 µg/plate ( TA 1535 (-S9-mix))
0, 1.4, 12, 37, 111, 333 µg/plate (TA 1535 (+S9-mix), TA 1537 (-S9-mix), TA 98 (+S9-mix) and TA 100 (-S9-mix))
0, 0.46, 1.4, 12, 37, 111 µg/plate (TA 1537 (+S9-mix) and TA 100 (+S9-mix))
0, 62, 185, 556, 1667, 5000 µg/plate (WP2 uvrA (+S9-mix))
Experiment 3 and 4:
0, 4.1, 12, 37, 111, 333, 1000 µg/plate ( TA 1535 (-S9-mix))

Second bacterial reverse mutation test
0, 20, 39, 78, 156, 313, 625 µg/plate (TA 1535 (-S9-mix))
0, 9.8, 20, 39, 78, 156, 313 µg/plate (TA 1535 (+S9-mix))
0, 4.9, 9.8, 20, 39, 78, 156 µg/plate (TA 1537, TA 98 (+S9-mix) and TA 100 (-S9-mix))
0, 39, 78, 156, 313, 625, 1250 µg/plate (TA 98 (-S9-mix))
0, 2.5, 4.9, 9.8, 20, 39, 78 µg/plate (TA 100 (+S9-mix))
0, 313, 625, 1250, 2500, 5000 µg/plate (WP2 uvrA)

Vehicle / solvent:

DMSO

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

ethylnitrosurea

other: 2-aminoanthracene, N-ethyl-N-nitrosourea

Remarks:

In the absence of S9-mix: sodium azide: TA 1535 and TA 100; 9-aminoacridine: TA 1537; 2-nitrofluorene: TA 98; N-ethyl-N-nitrosourea: WP2uvrA. In the presence of S9-mix: 2-aminoanthracene: TA 1535, TA 98, TA 100, WP2uvrA; benzo(a)pyrene: TA 1537

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48-72 hours at ca. 37 °C

NUMBER OF REPLICATIONS: All determinations were made in triplicate.

DETERMINATION OF CYTOTOXICITY
- Toxicity was defined as a reduction (by at least 50%) in the number of revertant colonies and/or a clearing of the background lawn of bacterial growth as compared to the negative (solvent) control and/or the occurrence of pinpoint colonies.

Evaluation criteria:

The study was considered valid if the mean colony counts of the vehicle control values of the strains were within the acceptable ranges, if the results of the positive controls met the criteria for a positive response, if no more than 5 % of the plates was lost through contamination or other unforeseen events and if at least three concentrations were non-toxic.

A test substance was considered to be positive in the bacterial gene mutation test if the mean number of revertant colonies on the test plates was increased in a dose-related manner or if a two-fold or greater increase was observed compared to the negative control plates. A clear positive response did not need to be verified. Marginally or weakly positive results should be verified by additional testing.

A test substance was considered to be negative in the bacterial gene mutation test if it showed neither a dose-related increase in the mean number of revertant colonies nor a reproducible positive response at any of the concentrations tested.

Positive results from the bacterial reverse mutation test indicate that a test substance induces point mutations by base pair substitutions or frameshifts in the genome of either Salmonella typhimurium and/or Escherichia coli. Negative results indicate that, under the test conditions used, the test substance is not mutagenic in the tested strains.Although most studies give clearly positive or negative results, in rare cases the data set may preclude making a definite judgement about the mutagenic potential of the test substance. Results may remain equivocal in this case.

Both numerical significance and biological relevance were considered together in the evaluation.

Statistics:

No statistical analysis was performed.

Key result

Species / strain:

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

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

Key result

Species / strain:

E. coli WP2 uvr A

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:

The first test included four experiments. In the first experiment, the test substance was found toxic for all Salmonella strains tested, both in the absence and presence of S9-mix. In addition, for strains TA 1535 (+S9-mix), TA 1537 (-/+ S9-mix), TA 98 (+S9-mix) and TA 100 (-/+S9-mix) less than three non-toxic concentrations could be evaluated. For strain TA 1535 (-/+ S9-mix), TA 100 (-S9-mix) and WP2UvrA (+S9-mix) the negative or positive control was outside the acceptable range. Therefore, the experiment was repeated for these strains.
In the second experiment the test substance was found toxic for all Salmonella strains tested, both in the absence and presence of S9-mix. But for all strains tested three or more non-toxic concentrations could be evaluated. For strain TA 1535 (-S9-mix), the negative control was outside the historical range. Therefore the experiment was repeated for this strain.
In the third experiment, for strain TA 1535 (- S9-mix), the negative control was outside the historical range. Therefore the experiment was repeated again. In the fourth experiment, the test substance was found toxic for strain TA 1535, in the absence of S9-mix, but at least three non-toxic concentrations could be evaluated. Based on the results of these four experiments, the first test was regarded as valid.

The second test included one experiment. In this fifth experiment, the test substance was found toxic for all Salmonella strains tested, both in the absence and presence of S9-mix, at at least one concentration, but for all strains tested three or more non-toxic concentrations could be evaluated. The mean numbers of his+ and trp+ revertant colonies of the negative controls in all strains used were within the acceptable range. In all strains, the positive controls gave the expected increase in the mean numbers of revertant colonies. Therefore, the second test was considered valid.

In all tests, toxicity was evidenced by a decrease in the mean number of revertants and/or a clearing of the background lawn of bacterial growth compared to the negative controls

In all experiments, a dose related precipitation of the test substance in the final treatment mix (top agar) was observed. Precipitation was observed in all mixtures at and above 556 μg/plate (first experiment), 333 μg/plate (second, third and fourth experiment) and 313 μg/plate (fifth experiment) with the unaided eye. In the first, second and fifth experiment, a precipitation of the test substance on the agar plates was observed. Precipitation was observed at and above 1667 μg/plate (first and second experiment) and at 5000 μg/plate (fifth experiment) with microscope only.

In both tests, in all strains tested, in both the absence and presence of S9-mix, the test substance did not induce a more than 2-fold and/or dose related increase in the mean number of revertant colonies compared to the background spontaneous reversion rate observed with the negative control.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

Under the test conditions (OECD 471, GLP) the results obtained in Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100, and in the Escherichia coli strain WP2 uvrA, in the absence and presence of the S9-mix, indicate that the test substance is not mutagenic.

Executive summary:

According to OECD guideline 471 and GLP, the test substance, dissolved in DMSO, was examined for its possible mutagenic activity in the bacterial reverse mutation test using Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and the Escherichia coli strain WP2uvrA, in the absence and presence of a liver fraction of Aroclor 1254-induced rats for metabolic activation (S9-mix). Two independent bacterial reverse mutation tests were performed. In the first test, four independent experiments were performed where six to ten concentrations of the test substance ranging from 4.1 to 5000 μg/plate were tested. In the second test, six concentrations of the test substance ranging from 2.5 to 1250 μg/plate were tested and five concentrations ranging from 313 to 5000 μg/plate for the Salmonella strains and WP2 UvrA strain, respectively. In all tests, negative controls (vehicle) and positive controls were run simultaneously with the test substance. In some of the experiments of the first test, the mean numbers of his+ and trp+ revertant colonies of the negative and positive controls in some of the strains appeared to be outside the acceptable range. Therefore, the experiments were repeated until the acceptance criteria were met. In the second test, the mean numbers of his+ and trp+ revertant colonies of the negative controls were within the acceptable range and the positive controls gave the expected increase in the mean numbers of revertant colonies in all strains. Therefore, both tests of this study were considered valid. In the first experiment of the first test, the test substance was found toxic to all strains tested, both in the absence and presence of S9-mix. For strains TA 1535 (+S9-mix), TA 1537 (-/+ S9-mix), TA 98 (+S9-mix) and TA 100 (-/+ S9-mix), less than three non-toxic concentrations could be evaluated, therefore the experiment was repeated for these strains (second, third and fourth experiment), which eventually resulted in a valid first test where at least three non-toxic concentrations could be evaluated. In the second test, toxicity was observed for all Salmonella strains tested, both in the absence and presence of S9-mix, but three or more non-cytotoxic concentrations could be evaluated. In all experiments, toxicity was evidenced by a decrease in the mean number of revertants and/or a clearing of the background lawn of bacterial growth compared to the negative controls. In both tests, in all strains tested, in both the absence and presence of S9-mix, the test substance did not induce a more than 2-fold and/or dose related increase in the mean number of revertant colonies compared to the background spontaneous reversion rate observed with the negative control. It is concluded that the results obtained in Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100, and in the Escherichia coli strain WP2 uvrA, in both the absence and presence of the S9-mix, indicate that the test substance is not mutagenic under the conditions used in this study.

Reference 2

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

Study period:

30 April 2015 (solubility test) and 12 June 2015 (last day of slide analysis)

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)

GLP compliance:

yes (incl. QA statement)

Remarks:

TNO Triskelion, Utrechtseweg 48, 3704 HE Zeist, The Netherlands

Type of assay:

in vitro mammalian chromosome aberration test

Species / strain / cell type:

lymphocytes: human

Details on mammalian cell type (if applicable):

Blood samples were obtained by venapuncture from young (24 and 31 years old) healthy, non-smoking individuals with no known recent exposures to genotoxic chemicals or radiation.

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver S9-mix

Test concentrations with justification for top dose:

First assay: 2.0, 3.9, 7.8, 15.6, 31.3, 62.5, 125, 250, 500, and 1000 μg/mL.
Second assay: 6.25, 12.5, 25, 50, 75, 100, 125, 150, 175, 200 μg/mL

Vehicle / solvent:

DMSO

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

mitomycin C

Remarks:

Mitomycin C: in the absence of S9, Cyclophosphamide: in the presence of S9

Details on test system and experimental conditions:

FIRST EXPERIMENT
In both the absence and presence of S9-mix, the treatment times were 4 hours (pulse treatment) and the harvest time of the cells was 24 hours after onset of treatment. Two hours before harvest colchicine was added and two hours later cells were harvested.

SECOND EXPERIMENT
In the absence of S9-mix the cultures were treated continuously with the test substance for 24 hours. Two hours before the end of treatment colcemid was added and two hours later cells were harvested.

SPINDLE INHIBITOR: Colchicine and colcemid
STAIN: Giemsa

NUMBER OF REPLICATIONS:
In all instances duplicate cultures were used.

NUMBER OF CELLS EVALUATED:
A number of 1000 stimulated lymphocytes (500 per slide) were examined in each culture to determine the percentage of cells in mitosis (mitotic index). On the basis of the results of the mitotic index scoring and the observations with respect to the number and quality of the metaphases, three concentrations of the test substance together with the negative (solvent) and positive controls were selected for chromosomal aberration analysis. For each treatment group, 300 well-spread metaphases per concentration (150 metaphases per culture and 75 metaphases per slide), each containing 46 centromeres, were analysed by microscopic examination for chromatid-type aberrations (gaps, breaks, fragments, interchanges), chromosome-type aberrations (gaps, breaks, minutes, rings, dicentrics), according to the criteria recommended by Savage (1975). If heavily damaged cells or cells with numerical aberrations (such as endoreduplicated cells or polyploid cells) were observed, these cells were recorded but not counted and included in the 300 analysed cells. The Vernier readings of all aberrant metaphases were recorded.

DETERMINATION OF CYTOTOXICITY
Method: mitotic index

Evaluation criteria:

- The study was considered valid if the positive controls demonstrated a statistically significantly increase in the number of aberrant cells and the solvent controls were within the historical range.
- There are several criteria for determining a positive response, such as a statistically significant concentration-related or a reproducible statistically significant increase in the number of aberrant cells at one or more test points compared with the concurrent solvent control. The test substance was also considered positive if the obtained results were outside the distribution of the historical solvent control data.
- A response was considered to be equivocal if the percentage of aberrant cells was statistically marginal higher than that of the solvent control (0.05- A test substance was considered to be negative if it demonstrated neither a statistically significant dose-related or reproducible statistically significant increase in the number of metaphases containing one or more aberrations, at any of the test points analysed.

Statistics:

- The number of metaphases containing one or more aberrations of the test substance treated groups were compared with those of the concurrent solvent controls using Fisher's exact test (one-sided). The difference was considered statistically significant when the p-value of the Fisher’s exact test was less than 0.05.
- Statistical methods were used as an aid in evaluating the test results but were not the only determining factor for a positive response. Both statistical methods and biological relevance of the results were considered together in the evaluation.

Key result

Species / strain:

lymphocytes: human

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:

RESULTS OF THE MITOTIC INDEX ANALYSIS
- In the first test, in the presence of S9-mix, the cultures of the dose levels (125, 62.5, 31.3, 15.6, 7.8 and 3.9 μg/mL) together with the cultures of the concurrent solvent control and positive control Cyclophosphamide were selected for mitotic index analysis. At the three highest dose levels (1000, 500 and 250 μg/mL), the test substance was severely cytotoxic to the cells, which was demonstrated by the absence of (stimulated) lymphocytes. The mitotic indices of lower dose levels (125, 62.5, 31.3 and 15.6 μg/ml) showed a decrease to 50%, 78%, 85% and 85%, respectively when compared to the mitotic index of the concurrent solvent control. The lowest dose levels (7.8 and 3.9 μg/mL) did not show a reduction of the mitotic index. The cells of the concentration of 2.0 μg/ml were stored without slide preparation. Based on the results of the mitotic index analysis, the cultures of three dose levels of the test substance (125, 62.5 and 7.8 μg/mL), together with the cultures of the concurrent solvent control and positive control (Cyclophosphamide) were selected for chromosomal aberration analysis.
In the absence of S9-mix, the cultures of all dose levels of the test substance (62.5, 31.3, 15.6, 7.8, 3.9 and 2.0 μg/mL) together with the cultures of the concurrent solvent control were selected for mitotic index analysis. At the four highest dose levels (1000, 500, 250 and 125 μg/mL), the test substance was severely cytotoxic to the cells, which was demonstrated by the absence of (stimulated) lymphocytes. The mitotic indices of lower dose levels (62.5 and 31.3 μg/mL) showed a dose related decrease to 48% and 79%, respectively when compared to the mitotic index of the concurrent solvent control. The lowest dose levels (15.6, 7.8, 3.9 and 2.0 μg/mL) did not show a reduction of the mitotic index. Based on the results of the mitotic index analysis, the cultures of three dose levels of the test substance (62.5, 31.3 and 15.6 μg/mL), together with the cultures of the concurrent solvent control were selected for chromosomal aberration analysis.
- In the second test, in the continuous treatment group without metabolic activation, the selected cultures of dose levels (125, 100, 75, 50, 25, 12.5 and 6.25 μg/mL) together with the cultures of the concurrent solvent control were selected for mitotic index analysis. At the four highest dose levels (200, 175, 150 and 125 μg/mL), the test substance was severely cytotoxic to the cells, which was demonstrated by the absence of (stimulated) lymphocytes (only cell debris present on the slides). The mitotic indices of lower dose levels (100, 75, 50, 25 and 12.5 μg/mL) showed a dose related decrease to 11%, 24%, 49%, 65% and 78%, respectively when compared to the mitotic index of the concurrent solvent control. The lowest dose level (6.25 μg/mL) did not show a reduction of the mitotic index. Based on the results of the mitotic index analysis, the cultures of three dose levels of the test substance (50, 25 and 6.25 μg/mL), together with the cultures of the concurrent solvent control and positive control (Mitomycin C) were selected for chromosomal aberration analysis.

RESULTS OF THE CHROMOSOMAL ABERRATION ANALYSIS
In both the first and second chromosomal aberration test, in both the presence and absence of a metabolic activation system (S9-mix), the test substance did not induce a statistically significant increase in the number of aberrant cells, at any of the concentrations and treatment conditions analysed, when compared to the number of aberrant cells found in the concurrent solvent control cultures.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

The test material was not clastogenic to cultured human lymphocytes under the conditions of this chromosome aberration test.

Executive summary:

In an in vitro mammalian cell chromosome aberration test, performed according to OECD 473 and in compliance with GLP, the test substance dissolved in DMSO was examined for its potential to induce chromosome aberrations in human lymphocytes in both the absence and presence of a metabolic activation system (S9 -mix). In the first chromosomal aberration test, in both the absence and presence of S9-mix, the treatment/harvesting times were 4/24 hours (pulse treatment). In both pulse treatment groups, a dose-related increase in cytotoxicity was observed. Due to severe cytotoxicity at the highest dose levels in both pulse treatment groups, as demonstrated by the absence of the lymphocytes, these dose levels could not be analysed. Based on the observed cytotoxicity, the following dose levels were selected for chromosomal aberration analysis: - Pulse treatment group with metabolic activation: dose levels of 7.8, 62.5 and 125 μg/mL, together with the cultures of the solvent and the positive control (Cyclophosphamide). - Pulse treatment group without metabolic activation: dose levels of 15.6, 31.3 and 62.5 μg/mL, together with the cultures of the solvent. In the second test, narrower spaced dose levels were tested. In the continuous treatment group the treatment/harvesting times were 24/24 hours. In this treatment group, a dose-related increase in cytotoxicity was observed. Three dose levels of the test substance (6.25, 25 and 50 μg/mL), together with the cultures of the solvent and the positive control (Mitomycin C) were analysed. In both chromosomal aberration tests, the numbers of cells with structural aberrations observed in the solvent control (1%-DMSO) cultures were within the historical range. Treatment with the positive controls Cyclophosphamide and Mitomycin C resulted in statistically significant increases in the numbers of metaphases containing one or more chromosomal aberrations, when compared to the numbers observed in the cultures treated with the solvent. This demonstrates the validity of the study. In both chromosomal aberration tests, the test substance did not induce a statistically significant increase in the number of aberrant cells, at any of the concentrations and treatment periods analysed, when compared to the number of aberrant cells observed in the solvent control cultures. From the results obtained in this chromosomal aberration test it is concluded that, the test substance was not clastogenic to cultured human lymphocytes, under the conditions used in this study.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Ames test:

The mutagenic activity of the substance in the bacterial reverse mutation test was evaluated in accordance with OECD guideline 471 and GLP. The test was performed in two independent experimentsusing Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and the Escherichia coli strain WP2uvrA, in the absence and presence of a liver fraction of Aroclor 1254-induced rats. The dose levels were selected based on observed cytotoxicity. Adequate negative and positive controls were included. In both tests, in all strains tested, in both the absence and presence of S9-mix, the test substance did not induce a more than 2-fold and/or dose related increase in the mean number of revertant colonies compared to the background spontaneous reversion rate observed with the negative control. It is concluded that the results obtained in Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100, and in the Escherichia coli strain WP2 uvrA, in both the absence and presence of the S9-mix, indicate that the test substance is not mutagenic under the conditions employed in this study.

In an in vitro mammalian cell chromosome aberration test, performed according to OECD 473 and in compliance with GLP, the test substance dissolved in DMSO was examined for its potential to induce chromosome aberrations in human lymphocytes in both the absence and presence of a metabolic activation system (S9 -mix). In the first chromosomal aberration test, in both the absence and presence of S9-mix, the treatment/harvesting times were 4/24 hours (pulse treatment). In both pulse treatment groups, a dose-related increase in cytotoxicity was observed. Due to severe cytotoxicity at the highest dose levels in both pulse treatment groups, as demonstrated by the absence of the lymphocytes, these dose levels could not be analysed. Based on the observed cytotoxicity, the following dose levels were selected for chromosomal aberration analysis: - Pulse treatment group with metabolic activation: dose levels of 7.8, 62.5 and 125 μg/mL, together with the cultures of the solvent and the positive control (Cyclophosphamide). - Pulse treatment group without metabolic activation: dose levels of 15.6, 31.3 and 62.5 μg/mL, together with the cultures of the solvent. In the second test, narrower spaced dose levels were tested. In the continuous treatment group the treatment/harvesting times were 24/24 hours. In this treatment group, a dose-related increase in cytotoxicity was observed. Three dose levels of the test substance (6.25, 25 and 50 μg/mL), together with the cultures of the solvent and the positive control (Mitomycin C) were analysed. In both chromosomal aberration tests, the numbers of cells with structural aberrations observed in the solvent control (1%-DMSO) cultures were within the historical range. Treatment with the positive controls Cyclophosphamide and Mitomycin C resulted in statistically significant increases in the numbers of metaphases containing one or more chromosomal aberrations, when compared to the numbers observed in the cultures treated with the solvent. This demonstrates the validity of the study. In both chromosomal aberration tests, the test substance did not induce a statistically significant increase in the number of aberrant cells, at any of the concentrations and treatment periods analysed, when compared to the number of aberrant cells observed in the solvent control cultures. From the results obtained in this chromosomal aberration test it is concluded that, the test substance was not clastogenic to cultured human lymphocytes, under the conditions used in this study.

Justification for classification or non-classification

Based on the results of the Ames test and the chromosome aberration test, the test substance does not have to be classified for mutagenicity in accordance with Regulation (EC) No. 1272/2008.