Registration Dossier
Registration Dossier
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 907-495-0 | CAS number: 198028-14-7
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data

Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
The available data from three in vitro assays show that the substance does not have a genotoxic potential.
Link to relevant study records
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From 2009-10-09 to 2009-12-16
- 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)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- Not applicable, chromosome aberration
- Species / strain / cell type:
- lymphocytes: human lymphocyte cultures
- Details on mammalian cell type (if applicable):
- not applicable
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 from Aroclor 1254 induced animals
- Test concentrations with justification for top dose:
- First experiment:
3+17 hour -S-9: 15.1, 130 and 200 µg/mL
3+17 hour +S-9: 54.9, 130 and 200 µg/mL
Second experiment:
20+0 hour -S-9: 125, 150, 175 and 200 µg/mL
3+17 hour +S-9: 140, 160, and 180 µg/mL - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle:the test substance was soluble in DMSO at concentrations up to approximately 80.10 mg/mL when heated to 80ºC with the aid of vortex mixing and ultrasonication. - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 4 Nitroquinoline 1-oxide (NQO) and cyclophosphamide (CPA)
- Remarks:
- no remarks
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium; in suspension
DURATION
- Preincubation period: no
- Exposure duration:
First experiment: 3 hours
Second experiment: 20 hours and 3 hours
- Expression time (cells in growth medium): no data
- Selection time (if incubation with a selection agent): not applicable
- Fixation time (start of exposure up to fixation or harvest of cells): 20 hours
SELECTION AGENT (mutation assays): no data
SPINDLE INHIBITOR (cytogenetic assays): no data
STAIN (for cytogenetic assays): the cells were stained for 5 minutes in filtered 4% (v/v) Giemsa in pH 6.8 buffer.
NUMBER OF REPLICATIONS: no data
NUMBER OF CELLS EVALUATED: no data
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index and percentage of cells in mitosis
OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreplication: yes
- Other: hyperdiploidy and structural aberrations - Evaluation criteria:
- For valid data, the test article was considered to induce clastogenic events if:
1. A proportion of cells with structural aberrations at one or more concentrations that exceeded the normal range was observed in both replicate cultures
2. A statistically significant increase in the proportion of cells with structural aberrations (excluding gaps) was observed (p < 0.05)
3. There was a concentration-related trend in the proportion of cells with structural aberrations (excluding gaps).
The test article was considered as positive in this assay if all of the above criteria were met.
The test article was considered as negative in this assay if none of the above criteria were met.
Results which only partially satisfied the above criteria were dealt with on a case by case basis. Evidence of a concentration-related effect was considered useful but not essential in the evaluation of a positive result [ ]. Biological relevance was taken into account, for example consistency of response within and between concentrations and/or between experiments, or effects occurring only at high or very toxic concentrations, and the types and distribution of aberrations. - Statistics:
- No data
- Key result
- Species / strain:
- lymphocytes: human lymphocyte cultures
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- True negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No marked changes in pH compared to the concurrent vehicle controls, were observed
- Effects of osmolality: No marked changes in osmolality compared to the concurrent vehicle controls, were observed
- Evaporation from medium: no data
- Water solubility: not soluble
- Precipitation: yes
- Other confounding effects: no data
RANGE-FINDING/SCREENING STUDIES: yes
COMPARISON WITH HISTORICAL CONTROL DATA: yes, the proportion of cells with structural aberrations in these cultures fell within current historical vehicle control (normal) ranges
ADDITIONAL INFORMATION ON CYTOTOXICITY: no data - Conclusions:
- The test substance did not induce chromosome aberration in cultured human peripheral blood lymphocytes in the absence and in the presence of activation system.
- Executive summary:
The test substance was tested in an in vitro cytogenetics assay using duplicate human lymphocyte cultures prepared from the pooled blood of three male donors. This study was performed in accordance with OECD Guideline 473 and Good Laboratory Practices.
A range-finding study was performed to select the test article concentrations for chromosome analysis, by evaluating the effect of the test substance on mitotic index. Treatments covering a broad range of concentrations, separated by narrow intervals, were performed both in the absence and presence of metabolic activation (S-9) from Aroclor 1254 induced animals.
In the first experiment, human lymphocyte cultures were treated with the test substance at concentrations of 15.1, 130 and 200 µg/mL in the absence of S9 and 54.9, 130 and 200 µg/mL in the presence of S9. In the second experiment, human lymphocyte cultures were treated with the test substance at concentrations of 125, 150, 175 and 200 µg/mL in the absence of S9 and 140, 160, and 180 µg/mL in the presence of S9.
Appropriate negative (vehicle) control cultures were included in the test system in both experiments under each treatment condition. The proportion of cells with structural aberrations in these cultures fell within current historical vehicle control (normal) ranges. 4-Nitroquinoline 1-oxide (NQO) and cyclophosphamide (CPA) were employed as positive control chemicals in the absence and presence of rat liver S-9 respectively.
Treatment of cells with the test substance in the absence and presence of S-9 in both experiments resulted in frequencies of cells with structural aberrations that were generally similar to those observed in concurrent vehicle control cultures for all concentrations analysed.
The only exception to this was observed in one culture at the lowest concentration (140 µg/mL) analysed in the presence of S-9 in Experiment 2, where the number of aberrant cells (excluding gaps) was markedly higher than the concurrent control values. There was therefore no evidence of reproducibility of this isolated effect between cultures or between experiments and the observation was not considered biologically relevant.
No increases in the total frequency of cells with numerical aberrations, which exceeded the concurrent controls and the normal ranges, were observed in cultures treated with the test substance in the absence and presence of S-9 in Experiments 1 and 2.
Therefore, the test substance did not induce chromosome aberrations in cultured human peripheral blood lymphocytes when tested to the limit of solubility in culture medium both in the absence and the presence of S-9.
- 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:
- From 2009-10-09 to january 2010
- 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)
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- The thymidine kinase (tk) gene
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: RPMI 10
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: no data
- Periodically "cleansed" against high spontaneous background: yes - Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9: Aroclor 1254 induced rat liver post mitochondrial fraction
- Test concentrations with justification for top dose:
- See table 7.6.1/1
In experiment 1, ten concentrations, ranging from 4.14 to 200 µg/mL, were tested in the absence and presence of S 9.
In Experiment 2, ten concentrations, ranging from 15.0 to 150 µg/mL in the absence of S-9 and from 15.0 to 200 ¿g/mL in the presence of S 9, were tested. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: no data - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: Methyl methane sulphonate (MMS); Benzo[a]pyrene (B[a]P)
- Remarks:
- no remarks
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Preincubation period: no data
For the cytotoxicity range-finder experiment, in the absence and presence of S 9, a 3 hour treatment incubation period was used.
- Exposure duration: 3 hours incubation at 37±1°C
- Expression time (cells in growth medium): Cultures were maintained in flasks for a period of 2 days during which the tk-/- mutation would be expressed.
- Selection time (if incubation with a selection agent): 12 to 14 days
- Fixation time (start of exposure up to fixation or harvest of cells): no data
SELECTION AGENT (mutation assays): 5 trifluorothymidine, TFT
SPINDLE INHIBITOR (cytogenetic assays): no data
STAIN (for cytogenetic assays): no data
NUMBER OF REPLICATIONS: in duplicate
NUMBER OF CELLS EVALUATED: no data
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency and relative total growth
OTHER EXAMINATIONS:
no data - Evaluation criteria:
- For valid data, the test article was considered to be mutagenic in this assay if:
1. The MF of any test concentration exceeded the sum of the mean control mutant frequency plus GEF
2. The linear trend test was positive.
The test article was considered as positive in this assay if both of the above criteria were met.
The test article was considered as negative in this assay if neither of the above criteria were met.
Results which only partially satisfied the assessment criteria described above were considered on a case-by-case basis. - Statistics:
- No data
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- True negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: yes, fluctuations in pH of more than one unit may be responsible for an increase in mutant frequencies
- Effects of osmolality: yes, changes in osmolality of more than 50 mOsm/kg may be responsible for an increase in mutant frequencies
- Evaporation from medium: no data
- Water solubility: no soluble
- Precipitation: yes, in the cytotoxicity Range-Finder Experiment, following the treatment incubation period, precipitate was observed at the highest concentration tested in the absence of S-9 (50.0 µg/mL) and at the highest three concentrations tested in the presence of S-9 (12.5 to 50.0 µg/mL).
- Other confounding effects: no data
RANGE-FINDING/SCREENING STUDIES: yes
COMPARISON WITH HISTORICAL CONTROL DATA:
ADDITIONAL INFORMATION ON CYTOTOXICITY: no data - Conclusions:
- The test substance did not induce mutation at the tk locus in the L5178Y mouse lymphoma cells in the presence and in the absence of activation system.
- Executive summary:
In an in vitro mammalian cell gene mutation test, the test substance was assessed for its ability to induce mutation at the tk locus (5-trifluorothymidine [TFT] resistance) in mouse lymphoma cells using a fluctuation protocol. This GLP study was performed according to OECD guideline 476 and the UKEMS guidelines.
The study consisted of a cytotoxicity Range-Finder Experiment followed by two independent experiments, each conducted in the absence and presence of metabolic activation by an Aroclor 1254 induced rat liver post-mitochondrial fraction (S-9).
In the cytotoxicity Range-Finder Experiment, six concentrations were tested in the absence and presence of S-9, ranging from 1.56 to 50 mg/mL (primarily limited by solubility in DMSO, heated to 37°C and using 1% v/v test article additions in culture medium).
In Experiments 1 and 2, the mutant frequencies of the concentrations plated were all less than the sum of the mean control mutant frequency plus the global evaluation factor (GEF, 126 mutants per 106viable cells), indicating a negative result. There were no statistically significant linear trends.
In Experiment 1, ten concentrations ranging from 4.14 to 200 mg/mL, were tested in the absence and presence of S-9. Two days after treatment, the highest concentrations selected to determine viability and TFT resistance were 84.5 mg/mL in the absence of S-9 and 130 mg/mL in the presence of S-9 (both limited by the appearance of post-treatment precipitate), which gave 82% and 106% RTG, respectively.
In Experiment 2, ten concentrations ranging from 15.0 to 150 mg/mL, in the absence of S-9 and from 15.0 to 200 mg/mL in the presence of S-9, were tested. Two days after treatment, the highest concentrations selected to determine viability and TFT resistance were 120 mg/mL in the absence of S-9 and 150 mg/mL in the presence of S-9 (both limited by the appearance of post-treatment precipitate), which gave 103% and 105% Relative Total Growth, respectively.
In Experiments 1 and 2, the Mutant Frequency (MF) values of the concentrations plated were all less than the sum of the mean control MF plus the GEF, indicating a negative result. There were no statistically significant linear trends.
Positive and negative controls were valid for both experiments.
Therefore, the test substance did not induce mutation at the tk locus of L5178Y mouse lymphoma. These conditions included treatments up to precipitating concentrations in two independent experiments in the absence and presence of a rat liver metabolic activation system (S-9).
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From 01-25-2010 to 03-16-2010
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- The histidine gene for strains S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
The tryptophan gene for strains E. coli WP2 uvr A pKM 101 - Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Details on mammalian cell type (if applicable):
- No data
- Additional strain / cell type characteristics:
- other: Histidine deficient
- Species / strain / cell type:
- E. coli WP2 uvr A pKM 101
- Details on mammalian cell type (if applicable):
- Not applicable
- Additional strain / cell type characteristics:
- other: Tryptophan deficient
- Metabolic activation:
- with and without
- Metabolic activation system:
- liver preparations (S9 mix) from rats treated with phenobarbital and 5,6-benzoflavone
- Test concentrations with justification for top dose:
- 1, 3, 10, 30, 100, 300 and 1000 µg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: the test substance was insoluble in water, ethanol and acetone - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: sodium azide; 9-Aminoacridine; 2-Nitrofluorene; 4-Nitroquinoline-1-oxide
- Remarks:
- In absence of S9 Mix
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 2-Aminoanthracene; Benzo[a]pyrene
- Remarks:
- In presence of S9 Mix
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Preincubation period:
First test: no preincubation period
Second test: at 37°C for 30 minutes
- Exposure duration: . all plates were incubated at approximately 37°C for ca 72 hours
- Expression time (cells in growth medium): not applicable
- Selection time (if incubation with a selection agent): not applicable
- Fixation time (start of exposure up to fixation or harvest of cells): not applicable
SELECTION AGENT (mutation assays):
- histidine production for S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- tryptophane production for E. coli WP2 uvr A pKM 101
NUMBER OF REPLICATIONS: Three Petri dishes were used for each treatment.
OTHER: no data - Evaluation criteria:
- If exposure to a test substance produces a reproducible increase in revertant colony numbers of at least twice (three times in the case of strains TA1535 and TA1537) the concurrent vehicle controls, with some evidence of a positive dose-response relationship, it is considered to exhibit mutagenic activity in this test system. No statistical analysis is performed.
If exposure to a test substance does not produce a reproducible increase in revertant colony numbers, it is considered to show no evidence of mutagenic activity in this test system. No statistical analysis is performed.
If the results obtained fail to satisfy the criteria for a clear “positive” or “negative” response, even after additional testing, the test data may be subjected to analysis to determine the statistical significance of any increases in revertant colony numbers. The statistical procedures used are those described by Mahon et al (1989) and are usually Dunnett’s test followed, if appropriate, by trend analysis.
Occasionally, these criteria may not be appropriate to the test data and, in such cases, the Study Director would use his/her scientific judgement. - Statistics:
- Dunnett's test
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A pKM 101
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no data
- Effects of osmolality: no data
- Evaporation from medium: no data
- Water solubility: no data
- Precipitation: Precipitate was observed on all plates containing 99422018 at 300 and 1000 µg/plate in both tests.
- Other confounding effects: no data
RANGE-FINDING/SCREENING STUDIES: no data
COMPARISON WITH HISTORICAL CONTROL DATA: no data
ADDITIONAL INFORMATION ON CYTOTOXICITY: no data - Remarks on result:
- other:
- Conclusions:
- The test substance showed no evidence of mutagenic activity in the bacterial system using Salmonella typhimurium and Escherichia coli.
- Executive summary:
The test substance was tested in a bacterial reverse mutation test according to OECD Guideline 471 (Bacterial Reverse Mutation Assay) and Good Laboratory Practices.
The strains S. typhimurium TA1535, TA1537, TA98 and TA100, and E. coli WP2 uvrA (pKM101) were treated with the test substance at concentrations of 1, 3, 10, 30, 100, 300 and 1000 µg in DMSO, with and without activation with liver preparations (S9 mix) from rats treated with phenobarbital and 5,6-benzoflavone.
A first test was performed without preincubation period whereas in a second test there was a preincubation period of 30 minutes. Vehicle and positive controls were performed in both tests.
In the first test, without preincubation period, no evidence of toxicity was obtained following exposure to the test substance. Precipitate was observed on all plates containing the test substance at 300 and 1000 µg/plate in both tests. A maximum exposure concentration of 1000 µg/plate was, therefore, selected for use in the second test.
No substantial increases in revertant colony numbers over control counts were obtained with any of the tester strains following exposure to the test substance at any concentration up to and including 1000 µg/plate in either the presence or absence of S9 mix.
In the second test, with 30 minutes of preincubation period, no evidence of toxicity was obtained following exposure to the test substance. Precipitate was observed on all plates containing the test substance at 300 and 1000 µg/plate in both tests.
No substantial increases in revertant colony numbers over control counts were obtained with any of the tester strains following exposure to the test substance at any concentration up to and including 1000 µg/plate in either the presence or absence of S9 mix.
The test substance showed no mutagenic activity in the bacteria system in the presence and in the absence of system of activation.
Referenceopen allclose all
Table 7.6.1/2: Experiment 1 – Results summary
Treatment |
Concentration (mg/mL) |
Cytotoxicity (%) |
% Cells with Chromosome Aberrations (Excluding Gaps) |
Historical(%)# |
Statistical significance |
|
|
|
|
|
|
|
|
|
|
3+17 hour -S-9 |
Vehiclea |
- |
2.00 |
0-3 |
- |
|
|
|
15.1 |
10 |
1.50 |
|
NC |
|
|
|
130 |
30 |
1.50 |
|
NC |
|
|
|
200 |
39 |
1.50 |
|
NC |
|
|
|
*NQO, 2.50 |
ND |
20.17 |
|
p=0.001 |
|
|
|
|
|
|
|
|
|
|
3+17 hour +S-9 |
Vehiclea |
- |
0.50 |
0-3 |
- |
|
|
|
54.9 |
23 |
1.50 |
|
NC |
|
|
|
130 |
4 |
0.00 |
|
NC |
|
|
|
200 |
17 |
1.00 |
|
NC |
|
|
|
*CPA, 12.5 |
ND |
15.76 |
|
p=0.001 |
|
|
|
|
|
|
|
|
|
|
aVehicle control,DMSO * Positive control #95thpercentile of the observed range NC = Not calculated ND = Not determined |
Table 7.6.1/3: Experiment 2 – Results summary
Treatment |
Concentration (mg/mL) |
Cytotoxicity (%) |
% Cells with Chromosome Aberrations (Excluding Gaps) |
Historical(%)# |
Statistical significance |
|
|
|
|
|
|
|
|
20+0 hour -S-9 |
Vehiclea |
- |
0.00 |
0-3 |
- |
|
|
125 |
9 |
1.00 |
|
NC |
|
|
150 |
17 |
0.50 |
|
NC |
|
|
175 |
47 |
0.50 |
|
NC |
|
|
200 |
34 |
0.50 |
|
NC |
|
|
*NQO, 2.50 |
ND |
40.82 |
|
p=0.001 |
|
|
|
|
|
|
|
|
3+17 hour +S-9 |
Vehiclea |
- |
1.50 |
0-3 |
- |
|
|
140 |
ND |
9.00 |
|
NC |
|
|
160 |
0 |
0.50 |
|
NC |
|
|
180 |
1 |
1.50 |
|
NC |
|
|
*CPA, 12.5 |
ND |
42.55 |
|
p=0.001 |
|
|
|
|
|
|
|
|
aVehicle control,DMSO * Positive control #95thpercentile of the observed range NC = Not calculated ND = Not determined |
Table 7.6.1/2: Experiment 1 (3 hour treatment in the absence and presence of S-9)
Treatment (µg/mL) |
-S-9 |
Treatment (µg/mL) |
+S-9 |
||||||||
|
%RTG |
MF§ |
|
%RTG |
MF§ |
||||||
0 |
100 |
82.9 |
0 |
100 |
66.9 |
||||||
4.14 |
105 |
80.4 |
4.14 |
95 |
90.0 |
||||||
6.37 |
107 |
79.8 |
6.37 |
100 |
65.6 |
||||||
9.80 |
108 |
80.5 |
9.80 |
104 |
81.7 |
||||||
15.1 |
98 |
62.6 |
15.1 |
108 |
64.6 |
||||||
23.2 |
128 |
62.9 |
23.2 |
104 |
72.9 |
||||||
35.7 |
118 |
53.3 |
35.7 |
100 |
67.4 |
||||||
54.9 |
103 |
68.5 |
54.9 |
114 |
75.0 |
||||||
84.5 P, PP |
82 |
109 |
84.5 P |
80 |
81.5 |
||||||
|
|
|
|
|
130 P, PP |
106 |
63.8 |
||||
Linear trend |
|
NS |
Linear trend |
|
NS |
||||||
MMS |
|
|
|
B[a]P |
|
|
|
||||
15.0 |
56 |
714 |
2.00 |
55 |
800 |
||||||
20.0 |
47 |
847 |
3.00 |
22 |
1220 |
||||||
|
|
|
|
|
|
|
|
|
|
|
|
Table 7.6.1/3: Experiment 2 (3 hour treatment in the absence and presence of S-9)
Treatment (µg/mL) |
-S-9 |
Treatment (µg/mL) |
+S-9 |
||||||||
|
%RTG |
MF§ |
|
%RTG |
MF§ |
||||||
0 |
100 |
61.0 |
0 |
100 |
71.4 |
||||||
40.0 |
99 |
52.7 |
45.0 |
106 |
42.4 |
||||||
50.0 |
99 |
62.4 |
60.0 |
108 |
53.9 |
||||||
60.0 |
103 |
49.9 |
75.0 |
99 |
67.3 |
||||||
70.0 |
107 |
49.1 |
90.0 P |
115 |
45.3 |
||||||
80.0 P |
94 |
58.9 |
105 P |
132 |
54.2 |
||||||
90.0 P |
101 |
65.3 |
120 P |
119 |
55.2 |
||||||
120 P,PP |
103 |
53.3 |
150 P, PP |
105 |
64.0 |
||||||
Linear trend |
|
NS |
Linear trend |
|
NS |
||||||
MMS |
|
|
|
B[a]P |
|
|
|
||||
15.0 |
44 |
190 |
2.00 |
84 |
448 |
||||||
20.0 |
32 |
364 |
3.00 |
43 |
675 |
||||||
|
|
|
|
|
|
|
|
|
|
|
|
MF: Mutant frequency
§: 5-TFT resistant mutants/106viable cells 2 days after treatment
% RTG: % Relative total growth
P: Precipitation observed at time of treatment
PP: Precipitation observed following treatment incubation period.
NS: Not significant
Table 7.6.1/1: Results obtained in the presence of metabolic activation: test 1
With metabolic activation
Strain |
Addition |
Concentration per plate |
Mean revertants per plate |
Standard Deviation |
Fold increase relative to vehicle |
Individual revertant |
|
|
|
|
|
|
|
TA98 |
DMSO |
- |
47.0 |
1.7 |
- |
46, 49, 46 |
|
test substance |
1 µg |
52.7 |
12.7 |
1.1 |
39, 64, 55 |
|
|
3 µg |
61.3 |
7.0 |
1.3 |
68, 62, 54 |
|
|
10 µg |
43.3 |
9.2 |
0.9 |
38, 38, 54 |
|
|
30 µg |
52.7 |
3.2 |
1.1 |
54, 49, 55 |
|
|
100 µg |
40.0 |
3.6 |
0.9 |
39, 44, 37 |
|
|
300 µg |
40.7 |
4.7 |
0.9 |
46 P, 39 P, 37 P |
|
|
1000 µg |
31.7 |
2.1 |
0.7 |
31 P, 34 P, 30 P |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
TA100 |
DMSO |
- |
170.7 |
8.1 |
- |
180, 166, 166 |
|
test substance |
1 µg |
192.0 |
8.7 |
1.1 |
202, 186, 188 |
|
|
3 µg |
190.7 |
23.5 |
1.1 |
214, 167, 191 |
|
|
10 µg |
163.3 |
16.1 |
1.0 |
145, 175, 170 |
|
|
30 µg |
166.7 |
39.1 |
1.0 |
170, 126, 204 |
|
|
100 µg |
175.7 |
17.6 |
1.0 |
174, 194, 159 |
|
|
300 µg |
166.0 |
25.2 |
1.0 |
189 P, 170 P, 139 P |
|
|
1000 µg |
132.3 |
10.5 |
0.8 |
122 P, 143 P, 132 P |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
TA1535 |
DMSO |
- |
24.0 |
3.5 |
- |
26, 26, 20 |
test substance |
1 µg |
22.0 |
4.0 |
0.9 |
22, 26, 18 |
|
|
|
3 µg |
21.7 |
2.1 |
0.9 |
21, 20, 24 |
|
|
10 µg |
24.7 |
4.0 |
1.0 |
20, 27, 27 |
|
|
30 µg |
20.7 |
1.2 |
0.9 |
20, 22, 20 |
|
|
100 µg |
19.3 |
1.2 |
0.8 |
18, 20, 20 |
|
|
300 µg |
15.3 |
3.1 |
0.6 |
16 P, 18 P, 12 P |
|
|
1000 µg |
17.7 |
1.5 |
0.7 |
19 P, 18 P, 16 P |
|
|
|
|
|
|
|
|
Key to Plate Postfix Codes |
|||
|
|
|
|
|
|
|
P |
Precipitate |
With metabolic activation
Strain |
Addition |
Concentration per plate |
Mean revertants per plate |
Standard Deviation |
Fold increase relative to vehicle |
Individual revertant |
|
|
|
|
|
|
|
TA1537 |
DMSO |
- |
34.3 |
4.6 |
- |
29, 37, 37 |
|
test substance |
1 µg |
32.0 |
10.6 |
0.9 |
44, 24, 28 |
|
|
3 µg |
33.7 |
5.0 |
1.0 |
33, 39, 29 |
|
|
10 µg |
34.0 |
8.7 |
1.0 |
38, 40, 24 |
|
|
30 µg |
39.7 |
2.1 |
1.2 |
39, 38, 42 |
|
|
100 µg |
32.0 |
9.5 |
0.9 |
23, 31, 42 |
|
|
300 µg |
24.7 |
3.8 |
0.7 |
22 P, 23 P, 29 P |
|
|
1000 µg |
22.7 |
11.6 |
0.7 |
35 P, 21 P, 12 P |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
WP2 uvrA |
DMSO |
- |
163.0 |
11.4 |
- |
176, 158, 155 |
(pKM101) |
test substance |
1 µg |
165.0 |
15.1 |
1.0 |
170, 148, 177 |
|
|
3 µg |
174.0 |
25.2 |
1.1 |
147, 178, 197 |
|
|
10 µg |
151.0 |
27.6 |
0.9 |
122, 177, 154 |
|
|
30 µg |
138.0 |
28.9 |
0.8 |
117, 171, 126 |
|
|
100 µg |
153.0 |
20.0 |
0.9 |
148, 175, 136 |
|
|
300 µg |
151.3 |
21.2 |
0.9 |
148 P, 174 P, 132 P |
|
|
1000 µg |
133.0 |
9.8 |
0.8 |
136 P, 141 P, 122 P |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
TA98 |
B[a]P |
5 µg |
406.7 |
45.7 |
8.7 |
431, 435, 354 |
TA100 |
AAN |
5 µg |
878.0 |
48.9 |
5.1 |
923, 826, 885 |
TA1535 |
AAN |
5 µg |
348.0 |
22.3 |
14.5 |
368, 352, 324 |
TA1537 |
B[a]P |
5 µg |
177.0 |
7.0 |
5.2 |
180, 182, 169 |
WP2 uvrA (pKM101) |
AAN |
10 µg |
476.7 |
14.5 |
2.9 |
460, 484, 486 |
|
|
|
|
|
|
|
Key to Positive Controls |
Key to Plate Postfix Codes |
|||
|
|
|
|
|
B[a]P AAN |
Benzo[a]pyrene 2-Aminoanthracene |
P |
Precipitate |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Reverse gene mutation assay
In a reverse gene mutation assay in bacteria performed according to the OECD test guideline No. 471 and in compliance with Good Laboratory Practice, strains TA 1535, TA 97, TA 98 and TA 100 of S. typhimurium and E. coli WP2 were exposed to the test substance in DMSO at concentrations of 0, 1, 3, 10, 30, 100, 300 and 1000 µg/plate in the presence and absence of mammalian metabolic activation (S9 mix) . In this study, there was no evidence of induced mutant colonies over background.
In vitro cytogenicity study in mammalian cells
In an in vitro chromosome aberration test performed according to the OECD guideline No. 473 and in compliance with Good Laboratory Practice, the human lymphocyte cultures were treated with the test substance in two experiments. In the first experiment the human lymphocyte cultures were treated with the test substance at concentrations of 0, 15.1, 130 and 200 µg/mL without metabolic activation and 0, 54.9, 130 and 200 µg/mL with metabolic activation (rat liver homogenate metabolising system). In the second experiment human lymphocyte cultures were treated with the test substance at concentrations of 125, 150, 175 and 200 µg/mL in the absence of S9 and 140, 160, and 180 µg/mL in the presence of S9. Two treatment conditions were used for the study: 4-hour exposure followed by a 17-hour expression period (with and without S9) and 20-hour exposure (without S9 only). No chromosome aberrations were induced by treatment with the test substance in cultured peripheral blood lymphocytes when tested to the limit of solubility in culture medium, both in the absence and the presence of S9. In this study, there was no evidence of in vitro cytogenicity of the test substance in mammalian cells.
In vitro gene mutation study in mammalian cells
In a mammalian cell gene mutation assay performed similarly to OECD 476, the test substance diluted in water was tested in the agar version of L5178Y mouse lymphoma assay. The test substance was tested in the mutagenesis assay over a range of concentrations from 4.14 to 200 mg/mL in the presence and absence of the S-9 activation. Both the non-activated and S-9 activated cultures did not produce significant increases in mutant frequency in comparison with that of the solvent control cultures.The test substance did not induce mutation at the tk locus of L5178Y mouse lymphoma cells.
Justification for classification or non-classification
Based on the results from three in vitro guideline compliant assays, the substance is not classified for genotoxicity according to regulation (EC) No. 1272/2008 and its subsequent amendments on classification, labeling and packaging (CLP) of substances and mixtures.
No further testing is required.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
