hexasodium 4-[3-(2-bromoprop-2-enamido)benzamido]-6-[(E)-2-[5-(2,3-dibromopropanamido)-2-sulfophenyl]diazen-1-yl]-5-hydroxynaphthalene-1,7-disulfonate 6-[(E)-2-[5-(2-bromoprop-2-enamido)-2-sulfophenyl]diazen-1-yl]-4-[3-(2-bromoprop-2-enamido)benzamido]-5-hydroxynaphthalene-1,7-disulfonate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Target substance was considered to be not mutagenic.

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:

1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

None

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

only four tester strains tested; no tester strain to detect cross-linking mutagens was included.

Principles of method if other than guideline:

Guideline followed

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

None

Target gene:

The strains are derived from S. typhimurium strain LT2 and due to a mutation in the histidine locus are histidine dependent. Additionally due to the "deep rough" (rfa-minus) mutation they possess a faulty lipopolysaccharide envelope which enables substances to penetrate the cell wall more easily. A further mutation causes a reduction in the activity of an excision repair system. The latter alteration includes mutational processes in the nitrate reductase and biotin genes produced in a UV-sensitive area of the gene named "uvrB-minus". In the strains TA 98, TA 100 and TA 102 the R-factor plasmid pKM 101 carries the ampicillin resistance marker. The strain TA 102 does not contain the uvrB-_mutation. Additionally TA 102 contains the multicopy plasmid pAQl, which carries the hisG428 mutation and a tetracyclin resistance gene. TA 102 contains the ochre mutation in hisG gene.

Species / strain / cell type:

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

Metabolic activation:

with and without

Test concentrations with justification for top dose:

Experiment 1: 33.3, 100.0, 333.3, 1000.0, 2500.0 and 5000.0 µg/plate
Experiment 2: 500.0, 1000.0, 2000.0, 3000.0, 4000.0 and 5000.0 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: A.bidest.
- Justification for choice of solvent/vehicle: The solvent was chosen because of its solubility properties.

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

methylmethanesulfonate

other: 4-nitro-o-phenylene-diamine

Details on test system and experimental conditions:

The bacterial strains were obtained from Dr. Heinz Träger, Knoll AG, D-6700 Ludwigshafen, F.R.G.

STORAGE:
The strain cultures were stored as stock cultures in ampoules with nutrient broth + 5 % DMSO in liquid nitrogen.

PRECULTURES:
From the thawed ampoules of the strains 0.5 ml bacterial suspension was transferred to 250 ml Erlenmeyer flasks containing 20 ml nutrient medium. This nutrient medium contains per litre:
8 g Merck Nutrient Broth
5 g NaCl
The bacterial culture was incubated in a shaking water bath for 10 hours at 37 °C.

SELECTIVE AGAR:
2.0 % Vogel-Bonner-Glucose-Minimal-Agar was used as selective agar. Each petri dish was filled with 20 ml of this nutrient medium. Sterilisations were performed at 121 °C in an autoclave.

OVERLAY AGAR:
The overlay agar contains per litre:
6.0 g Merck Agar Agar
6.0 g NaCl
10.5 mg L-histidine x HCl x H20
12.2 mg biotin
Sterilisations were performed at 121 °C in an autoclave.

MAMMALIAN MICROSOMAL FRACTION S9 MIX:
S9 (Preparation by C C R):
The S9 liver microsomal fraction was obtained from the liver of 8-12 weeks old male Wistar rats, strain WU (SAVO-Ivanovas, med. Versuchstierzuchten GmbH, D-7964 Kisslegg, F.R.G.; weight approx. 150 - 200 g) which received a single i.p. injection of 500 mg/kg b.w. Aroclor 1254 (Antechnika, D-7500 Karlsruhe, F.R.G.) in olive oil 5 days previously.

After cervical dislocation the livers of the animals were removed, washed in 150 mM KCl and homogenised. The homogenate, diluted 1+3 in KCl was centrifuged cold at 9,000 g for 10 minutes. A stock of the supernatant containing the microsomes was frozen in ampoules of 2, 3 or 5 ml and stored at -70 °C. Small numbers of the ampoules are kept at -20 °C for only several weeks before use. The standardisation of the protein content was made using the analysis kit of Bio-Rad Laboratories, D-8000 München: Bio-Rad protein assay, Catalogue 500 000 6.

The protein concentration in the S9 preparation was 31.6 mg/ml (lot 060792).

S9 MIX:
Before the experiment an appropriate quantity of S9 supernatant was thawed and mixed with S9 cofactor solution. The amount of S9 supernatant was 15 % v/v. The composition of the cofactor solution was concentrated to yield the following concentrations in the S9 mix:
8 mM MgCl2
33 mM KCl
5 mM glucose-6-phosphate
5 mM NADP
in 100 mM sodium-ortho-phosphate-buffer, pH 7.4.
During the experiment the S9 mix was stored in an ice bath. The S9 mix preparation was performed according to Ames et al.

PRE-EXPERIMENT FOR TOXICITY:
To evaluate the toxicity of the test article a prestudy was performed with strains TA 98 and TA 100. 8 concentrations were tested for toxicity and mutation induction with each 3 plates. The experimental conditions in this pre-experiment were the same as described below for the experiment I (plate incorporation
test).

Toxicity of the test article may be evidenced by a reduction in the number of spontaneous revertants, a clearing of the bacterial background lawn, or by degree of survival of treated cultures.

DOSE SELECTION:
According to the results of the pre-experiment the concentrations applied in the main experiments were chosen.
Experiment I was performed with the normal concentration grading. Due to the mutagenic effect obtained in experiment I, experiment II was performed as well as a plate incorporation assay. Additionally the concentration grading was designed to explore the linear part of the dose-response curve obtained in experiment I. Therefore, a more narrow concentration range (between 1000.0 and 5000.0 µg/plate) was selected in experiment II.

The maximum concentration was 5000.0 µg/plate in experiment I and II. The concentration range included two logarithmic decades in experiment I and one logarithmic decade in experiment II. In this study six adequately spaced concentrations were tested. Two independent experiments were performed.

Evaluation criteria:

A test article is considered as positive if either a dose related and reproducible increase in the number of revertants or a significant and reproducible increase for at least one test concentration is induced.

A test article producing neither a dose related and reproducible increase in the number of revertants nor a significant and reproducible positive response at any one of the test points is considered non-mutagenic in this system.

A significant response is described as follows:

A test article is considered as mutagenic if in strain TA 100 and TA 102 the number of reversions is at least twice as high and in strains TA 1535, TA 1537, and TA 98 it is at least three times higher as compared to the spontaneous reversion rate.

Also, a dose-dependent and reproducible increase in the number of revertants is regarded as an indication of possibly existing mutagenic potential of the test article regardless whether the highest dose induced the above described enhancement factors or not.

Statistics:

No appropriate statistical method is available

Key result

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

Toxic effects, evidenced by a reduction in the number of revertants, occurred in strain TA 1537 (without S9 mix) and in strain TA 102 (with S9 mix) at the highest investigated dose in experiment I.

The plates incubated with the test article showed normal background growth up to 5000.0 µg/plate with and without S9 mix in all strains used.

Up to the highest investigated dose a significant, dose-dependent increase in the number of revertants was observed in the strains TA 1535 and TA 100 with and without metabolic activation in experiment I and II.

No substantial increases in revertant colony numbers were observed following treatment with FAT 92'348/A (Lanasol Rot 2G roh trocken) at any dose level, either in the presence or absence of metabolic activation (S9 mix) in the strains TA 1537, TA 98 and TA 102.

A slight increase in revertant colony numbers was found in strain TA 98 (without S9 mix) at 5000.0 µg/plate in experiment I and in strain TA 102 at 3000.0 µg/plate (with S9 mix) in experiment II. These results are considered not to be biologically relevant since they could not be reproduced in the independent experiment.

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

Remarks on result:

other: all strains/cell types tested

None

Conclusions:

FAT 92'348/A (Lanasol Rot 2G roh trocken) is considered to be mutagenic in this Salmonella typhimurium reverse mutation assay.

Executive summary:

This study was performed to investigate the potential of FAT 92348/A (Lanasol Rot 2G roh trocken) to induce gene mutations according to the plate incorporation test using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100, and TA 102.

This study followed the procedures indicated by the following internationally accepted guidelines and recommendations:

First Addendum to OECD Guidelines for Testing of Chemicals - Section 4, No. 471, "Salmonella typhimurium, Reverse Mutation Assay", adopted May 26, 1983 and EEC Directive 79/831, Annex V, B 14. The study was performed in compliance with GLP.

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

Experiment I : 33.3; 100.0; 333.3; 1000.0; 2500.0 and 5000.0 µg/plate

Experiment II: 500.0; 1000.0; 2000.0; 3000.0; 4000.0 and 5000.0 µg/plate

Toxic effects, evidenced by a reduction in the number of revertants, occurred in strain TA 1537 (without S9 mix) and in strain TA 102 (with S9 mix) at the highest investigated dose in experiment I.

The plates incubated with the test article showed normal background growth up to 5000.0 µg/plate with and without S9 mix in all strains used. Up to the highest investigated dose a significant dose-dependent increase in the number of revertants was observed in the strains TA 1535 and TA 100 with and without metabolic activation in experiment I and II. Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies.

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test article induced point mutations by base pair changes in the genome of the strains TA 1535 and TA 100. Therefore, FAT 92'348/A (Lanasol Rot 2G roh trocken) is considered to be mutagenic in this Salmonella typhimurium reverse mutation assay.

Reference 2

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

Study period:

1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

None

Qualifier:

according to guideline

Guideline:

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

Deviations:

not specified

Principles of method if other than guideline:

Guideline followed

GLP compliance:

yes (incl. QA statement)

Type of assay:

mammalian cell gene mutation assay

Specific details on test material used for the study:

None

Species / strain / cell type:

mammalian cell line, other:

Metabolic activation:

with and without

Test concentrations with justification for top dose:

Cytotoxicity test:
A preliminary range finding test was run assessing cytotoxicity. FAT 92348/A was tested at concentrations up to 5000.0 µg/ml. This concentration represents the test limit dose.

Mutagenicity test with metabolic activation:
The original experiment was performed at the following concentrations: 2.59, 7.78, 23.33 and 70.0 µg/ml.

Mutagenicity test without metabolic activation:
The original experiment was performed at the following concentrations: 25.93, 77.78, 233.33 and 700.0 µg/ml.

Vehicle / solvent:

Ham's F10 culture medium.

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Details on test system and experimental conditions:

The test system allows the detection of base-pair substitutions, frameshift mutations and deletions induced by the test substance or by its metabolites. Mutagenic effects are manifested by the appearance of cells resistant to 6-TG and can be quantified by comparison of the numbers of 6-TG resistant colonies in the treated and control cultures. To ensure that any mutagenic effect of metabolites of the test substance found in mammals is also detected, an experiment is performed, in which the metabolic turnover of the test material is simulated in vitro by the addition of an activation mixture to the cell cultures containing rat-liver post mitochondrial supernatant (S9 fraction) and cofactors.

Maintenance of the cell line:
V79 Chinese hamster cells were originally derived from embryonic lung tissue. The cells were cultured in Ham's F10 medium supplemented with 10% pre-tested foetal calf serum, 100 U/ml penicillin and 100 fig/ml streptomycin in tissue culture (plastic) flasks. The humidity in the incubator was adjusted to >85 % rH, the air was enriched to 5 ±2.0 Vol% CO2 and the temperature was 37±1 °C. Twice per week the growth medium was replaced by fresh one. The laboratory cultures were passaged weekly in low number (about 5x10E4 cells per 175 cm²) to keep the level of spontaneous mutants low and to prevent the cells of reaching a stationary phase of cell growth. Large stocks of the V79 cell line have been stored in liquid nitrogen allowing the repeated use of the same cell culture batch in experiments. Consequently, the parameters of the experiments remain similar because of the reproducible characteristics of the cells. The frozen cell suspension contains 10 % dimethylsulfoxide (DMSO). All stock cells were cultured in cleansing medium for three days to purge the cultures of existing hprt" mutants. Cleansing medium was growth medium supplemented with 3 uM aminopterin. The cells have a stable karyotype with a modal chromosome number of 22±1. All stock cells were checked for mycoplasma contamination, using the Hoechst-Dye staining method or the 6-MPDR method, before being frozen. Thawed stock culture cells are kept not longer than for twelve passages (three months) in culture.

Preparation and composition of the metabolic activation mixture:
Rat-liver post mitochondrial supernatant (S9 fraction) was prepared in advance from male RAI rats (Tif: RAIffSPF]), reared at the Animal Farm of CIBA-GEIGY, Sisseln, Switzerland. The animals were treated with Aroclor 1254 (500 mg/kg, i.p.) 5 days prior to sacrifice. The livers were homogenized with 3 volumes of 150 mM KCl. The homogenate was centrifuged at 9000x g for 15 minutes and the resulting supernatant (S9 fraction) was stored at approximately minus 80 °C for no longer than one year. The protein content of the S9 fraction was 36.64 mg/ml. S9 fraction was thawed immediately before use. The S9 mixture was prepared just prior use in an activation experiment and kept on ice. Unused portions of S9 fraction and S9 mixture were discarded and not saved for another experiment. The S9 mixture was immediately sterilised by filtration through a 0.45 urn filter. The activation mixture was added to the medium at a concentration of 10 % in both the cytotoxicity test and the mutagenicity test and the final concentration of S9 fraction was 2.5 % during the treatment.

Evaluation criteria:

Assay acceptance criteria:
• The results of the experiments should not be influenced by a technical error, contamination or a recognized artifact.
• From each experiment, at least three concentrations of the test substance, one positive and one solvent control should be evaluated.
• The mutant frequency of the solvent controls (spontaneous mutant frequency) should not exceed 35x10E-6.
• The positive control should fulfil the criteria for a mutagenic substance.
• The highest concentration of the test substance applied in the mutagenicity test should either reduce the viable cells by about 50-90 % or correspond to the test substance's solubility limit (precipitates in the culture). In case of non-toxic freely soluble compounds the highest tested concentration will be 5 mg/ml. In special cases the highest concentration can be determined by the sponsor.

Assay evaluation criteria:
All mutant frequencies are normalized to a virtual cloning efficiency of 100 % at the end of the expression period. If the cloning efficiency of the viability cultures is lower than 15 %, the corresponding mutant frequency is usually not calculated, owing to the high statistical insignificance of the result. For every concentration a mean mutant factor, which is defined as the ratio of the mean mutant frequencies of the treated cultures with the mean mutant frequencies of the solvent control cultures, will be calculated.

Criteria for a positive response:
The test substance will be considered to be mutagenic if:
• The assay is valid (see assay acceptance criteria)
• The mutant frequency at one or more concentrations is significantly greater than that of the negative control and the number of normalized mutant clones in the treated and untreated cultures differs by more than 20.
• There is a significant dose-relationship as indicated by the linear trend analysis.
• The effects described above are reproducible.

Statistics:

Statistical significance of mutant frequencies was carried out according to the UKEMS guidelines.

Key result

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

Toxicity:
In the preliminary toxicity test with and without metabolic activation 12 concentrations of FAT 92348/A were tested. The concentrations selected ranged from 2.44 to 5000.0 µg/ml and separated by 2-fold intervals.
In the part with metabolic activation FAT 92348/A exerted a complete growth inhibitory effect down to the concentration of 625.0 µg/ml. The next lower concentration (312.5 µg/ml) produced an acute growth inhibition of 98.88 %. A similar effect was also seen at the next two lower concentrations. The concentration of 39.06 µg/ml then showed an acute inhibition of growth of 27.72 %. In the part without metabolic activation FAT 92348/A was completely growth inhibiting down to the concentration of 1250.0 µg/ml. The next lower concentration (625.0 µg/ml) revealed an acute growth inhibitory effect of 60.46 %.

Accordingly, four concentrations were selected for the original experiment ranging from 2.59 to 70.0 µg/ml and from 25.93 to 700.0 µg/ml in the presence and absence of metabolic activation, respectively. In the part with metabolic activation, the growth inhibition determined after treatment at the highest concentration of 70.0 µg/ml showed a mean value of 23.46 %. After expression, at this concentration a growth inhibition of 13.88 % was found. In the absence of metabolic activation the highest concentration proved completely growth inhibiting. The mean growth inhibitory effect determined after treatment was 84.18 % at the next lower concentration. At this concentration after the expression period the determined cytotoxicity revealed a mean value of 10.04 %. Since no pronounced growth inhibitory response was obtained in the part with metabolic activation, the concentration range was slightly increased in the confirmatory experiment ranging from 9.38 to 75.0 µg/ml. In the part without metabolic activation a slightly decreased concentration range of 81.25 to 650.0 µg/ml was selected, since the highest concentration proved completely growth inhibiting in the original experiment. In the presence of metabolic activation the mean acute cytotoxicity at the highest concentration was 55.66 %. The mean growth inhibitory effect after expression revealed 16.70 %). In the part without activation, the mean growth inhibition at the highest concentration of 650.0 µg/ml was 97.04%. After the expression period a value of 69.70 % was observed.

Mutagenicity:
In the presence and absence of metabolic activation, no biologically significant increase in mutan frequency was observed at any concentration level of of FAT 92348/A tested in the original or the confirmatory experiment in comparison with the negative control. The only statistically significant difference in mutant frequency observed in the confirmatory experiment without metabolic activation at the highest concentration is considered to be of no biological relevance, since at this concentration excessive toxicity of 97.04 % was observed after treatment.

The positive controls induced a clear increase in mutant frequency.

Remarks on result:

other: all strains/cell types tested

None

Conclusions:

FAT 92348/A and its metabolites did not show any mutagenic activity in this forward mutation system.

Executive summary:

Following OECD, EPA, EEC guidelines and in accordance with GLP, FAT 92348/A ca. 100 % purity, was tested for mutagenic effects on V79 Chinese hamster cells in vitro. The test substance was dissolved in Ham's F10 culture medium.

The cells were treated in the experiments with metabolic activation for 5 hours and in the experiments without metabolic activation for 21 hours. The results of each experiment were confirmed in a second and independent experiment (confirmatory experiment).

Cytotoxicity test

A preliminary range finding test was run assessing cytotoxicity. FAT 92348/A was tested at concentrations up to 5000.0 µg/ml. This concentration represents the test limit dose. In the part with metabolic activation, the test material was completely growth inhibiting down to the concentration of 625.0 ug/ml. At the next lower concentration of 312.5 ug/ml an acute growth inhibiting effect of 98.88% could be seen. A similar growth inhibiting effect was also observed at the next two lower concentrations. The concentration of 39.06 ug/ml revealed then an acute inhibition of growth of 27.72 %. Without metabolic activation treatment with FAT 92348/A proved growth inhibiting down to the concentration of 1250.0 µg/ml. The next lower concentration of 625.0 ug/ml revealed an acute inhibition of growth of 60.46 %. Accordingly, 70.0 µg/ml with and 700.0 µg/ml without metabolic activation were chosen as highest concentrations for the first mutagenicity assay.

Mutagenicity test with metabolic activation

The original experiment was performed at the following concentrations: 2.59, 7.78, 23.33 and

70.0 ug/ml. The mean growth inhibiting values found at the highest concentration after treatment

and expression were 23.46 %* and 13.88 %* respectively. In the confirmatory experiment the concentrations applied were 9.38, 18.75, 37.5 and 75.0 µg/ml The highest concentration revealed a mean acute growth inhibition of 55.66 %*. The mean growth inhibitory effect after the expression period was 16.70 %*. N-Nitrosodimethylamine (DMN, 1.0 ul/ml) was used as positive control. In both experiments comparison of the number of mutant colonies in the controls and in the cultures treated with the various concentrations of the test substance revealed no relevant increase of the mutant frequencies as determined by the screening with 6-Thioguanine (6-TG). In both experiments comparison of the number of mutant colonies in the controls and in the cultures treated with the various concentrations of the test substance revealed no relevant increase of the mutant frequencies as determined by the screening with 6-TG.

Mutagenicity test without metabolic activation

The original experiment was performed at the following concentrations: 25.93, 77.78, 233.33 and 700.0 µg/ml. The highest concentration proved completely toxic. The mean growth inhibition values found at the next lower concentration (233.33 µg/ml) after treatment and expression were 84.18 %* and 10.04 %* respectively. In the confirmatory experiment the concentrations applied were 81.25, 162.5, 325.0 and 650.0 µg/ml. The highest concentration revealed a mean acute growth inhibitory effect of 97.04 %*. The mean growth inhibition after the expression period was 69.07%*. Ethylmethansulfonate (EMS, 0.3 µl/ml) was used as positive control. In both experiments comparison of the number of mutant colonies in the controls and in the cultures treated with the various concentrations of the test substance revealed no relevant increase of the mutant frequencies as determined by the screening with 6-TG.

Based on the results of two independently performed experiments and under the given experimental conditions, it is concluded that FAT 92348/A and its metabolites did not show any mutagenic activity in this forward mutation system.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

FAT 92348/B was evaluated for genetic toxicity in a bacterial reverse mutation assay and an in vitro mammalian chromosomal aberration assay. However, a weak mutagenic effect was observed in the reverse mutation assay with FAT 92348/B. Hence to assess the genetoxic potential of FAT 92348/B, genetic toxicity data with a source substance FAT 40007/B is used.

In the reverse mutation assays (Ames Test) using bacteria (Salmonella typhimurium), both substances FAT 92348/A and FAT 40007/B exerted mutagenic effects.

 

In the in vitro mammalian chromosome aberration test, FAT 40007/B TE induced structural chromosome aberrations in the V79 Chinese hamster cell line.

 

In the in vitro mammalian cell gene mutation tests (HPRT-Locus) in Chinese hamster V79 cells, both FAT 92348/A and FAT 40007/B did not show any mutagenic activity.

To confirm the mutagenicity of FAT 40007/B, two in vivo tests were performed.

In the Micronucleus test, the test article did not induce micronuclei in the bone marrow cells of the mouse.

In in vivo UDS Test, the test item did not induce DNA damage leading to increased repair synthesis in the hepatocytes of the treated rats.

On the basis of available data, source substance FAT 40007/B was considered to be neither mutagenic nor clastogenic. Based on these results, the target substance FAT 92348/B (Reactive Red 116) is also considered to be neither mutagenic nor clastigenic based on the structure similarity of both test substances.

Justification for classification or non-classification

As discussed above, target substance was considered to be not genotoxic. Hence, the results do not warrant classification according to EU CLP.