Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
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: 268-028-8 | CAS number: 67990-05-0
- 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
Gene mutation in bacteria in vitro (OECD 471): negative with and without metabolic activation
Chromosome aberration in vitro (OECD 473): negative with and without metabolic activation (read-across from analogue substance Permanent Carmin FBB 02 [N-(4-chloro-2,5-dimethoxyphenyl)-3-hydroxy-4-[[2-methoxy-5-[(phenylamino)carbonyl]phenyl]azo]naphthalene-2-carboxamide])
Gene mutation in mammalian cells in vitro (OECD 476): negative with and without metabolic activation (read-across from analogue substance Permanent Carmin FBB 02 [N-(4-chloro-2,5-dimethoxyphenyl)-3-hydroxy-4-[[2-methoxy-5-[(phenylamino)carbonyl]phenyl]azo]naphthalene-2-carboxamide])
Link to relevant study records
- 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:
- From 13 JUL 2011 to 12 AUG 2011
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study (OECD TG471) with Prival modification for azo-dyes, GLP compliant
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: Commission Directive 2000/32/EC, L1362000, Annex 4D
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Specific details on test material used for the study:
- In October 2011 the EU commission published a recommendation (2011/696/EU) on the definition of nanomaterials. From the results of analyses it is concluded that the registered substance C.I.Pigment Red 269 falls within the boundaries of the nanomaterial definition. Hence, studies were performed using the nanomaterial.
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
- Metabolic activation:
- with and without
- Metabolic activation system:
- rat liver S9 or hamster liver S9
- Test concentrations with justification for top dose:
- Preliminary toxicity test (Plate incorporation method): 0, 0.075, 0.25, 0.75, 2.5, 7.5, 25, 75, 250, 750, 2500 µg/plate (only tested in strain TA 100)
Mutation test I and II (Pre-incubation method): 0, 25, 75, 250, 750, 2500 µg/plate
The maximum dose level was dictated by the solubility of the test item. - Vehicle / solvent:
- - Vehicle/ Solvent used: Dimethyl formamide
- Justification for choice of solvent/ vehicle: solubility properties - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: N-ethyl-N´-nitro-N-Nitrosoguanidine (TA 100, TA 1535), 9-Aminoacridine (TA 1537), 4-Nitroquinoline-1-oxide (TA 98), Mitomycin C (TA 102)
- Remarks:
- without metabolic activation
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene (TA 100, TA 1535, TA 1537), Benzo[a]pyren (TA 98), 1,8-Dihydroxyanthraquinone (TA 102)
- Remarks:
- with metabolic activation (rat liver S9 mix)
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: congo red (TA98, TA 100)
- Remarks:
- with metabolic activation (hamster liver S9 mix)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION:
Preliminary toxicity test: plate incorporation method with induced rat liver S9 mix (induction with phenobarbital/beta-naphthoflavone)
Mutation test I and II: pre-incubation method with non-induced hamster liver S9 mix.
DURATION:
-preincubation period (experimentII): 30 °C, for 30 min
- exposure duration: 48 h at 37 °C in the dark
NUMBER OF REPLICATIONS: 3 plates per strain and dose level including the controls for the mutation test I and II. - Evaluation criteria:
- There are several criteria for determining a positive result. Any , one, or all of the following can be used to determine the overall result of the study:
1. A dose-related increase in mutant frequency over the dose range tested.
2. A reproducible increase at one or more concentrations.
3. Biological relevance against in-house historical control ranges.
4. Statistical analysis of data as determined by UKEMS.
5. Fold increase greater than two times the concurrent solvent control for any tester strain (especially if accompanied by an out-of-historical range response)
A test item will be considered non-mutagenic (negative) in the test system if the above criteria are not met.
Although most experiments will give clear positive or negative results, in some instances the data generated will prohibit making a definite judgement about test item activity. results of this type will be reported as equivocal. - Statistics:
- Arithmetic mean and standard deviation were calculated .
- 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:
- valid
- 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:
- valid
- Positive controls validity:
- valid
- 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:
- valid
- Positive controls validity:
- valid
- 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:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 102
- 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:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: occured at concentrations of 750 µg/plate and higher in the preliminary toxicity test as well as in the mutation test I and II.
Nevertheless undissolved particles had no influence on the data recording. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'. Remarks: Mutation test I and II
- Conclusions:
- Interpretation of results (migrated information):
negative
The test item did not exert mutagenic activity in the reverse bacterial mutation assay (plate incorporation assay and pre-incubation assay according to Prival) with and without metabolic activation. - Executive summary:
Mutagenic activity of the test item was investigated in Salmonella typhimurium strain TA100 with (induced rat liver S9 mix) and without metabolic activation at concentrations of 0, 0.075, 0.25, 0.75, 2.5, 7.5, 25, 75, 250, 750 and 2500 µg/plate using the plate incorporation assay.
Due to the test items characteristic as an azo-dye the test was also conducted using the Prival modification, i.e. testing S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 in the preincubation assay with uninduced hamster liver S9 mix for metabolic activation. This test was performed using the following concentrations: 0, 25, 75, 250, 750 and 2500 µg/plate.
The test item did not reveal any mutagenic activity under the conditions tested. The appropriate reference mutagenes showed distinct positive mutagenic effects.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- key study
- Study period:
- From 24 OCT 2007 to 15 FEB 2008
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study (OECD 473), GLP compliant
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: Commission Directive 2000/32/EC, L1362000, Annex 4A, dated May 19, 2000
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: supplemented MEM
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- liver S9 extract from phenobarbital/beta-naphthoflavone induced Wistar rats
- Test concentrations with justification for top dose:
- without metabolic activation (P: precipitation occured)
Experiment 1: 1.3, 2.7 (P), 5.3 (P), 10.6 (P), 21.3 (P), 42.5 (P) µg/ml
Experiment 2a: 0.3, 0.6, 1,3, 2.5, 5.0 (P), 10.0 (P) µg/ml
Experiment 2b: 0.3, 0.6, 1,3, 2.5 (P), 5.0 (P), 10.0 (P) µg/ml
with metabolic activation (P: precipitation occured)
Experiment 1: 1.3, 2.7 (P), 5.3 (P), 10.6 (P), 21.3 (P), 42.5 (P), 85.0 (P), 170 (P) µg/ml
Experiment 2: 0.3, 0.6, 1.3, 2.5, 5.0 (P), 10.0 (P) µg/ml
Not all test groups were evaluated (for evaluation points see executive summary) - Vehicle / solvent:
- - Vehicle used: DMSO
- Justification for choice of solvent/vehicle: solubility properties and low toxicity to cell cultures - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- without metabolic activation
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Remarks:
- with metabolic activation
- Details on test system and experimental conditions:
- Two independent experiments were performed:
Experiment 1 with an exposure duration of 4 h without and with metabolic activation and chromosome preparation 18 h after start of treatment
Experiment 2 with an exposure duration of 18 and 28 h (each without metabolic activation) and chromosome preparation 18 h and 28 h after start of treatment, respectively as well as an exposure duration of 4 h (with metabolic activation) and chromosome preparation at 28 h after start of treatment. Two parallel cultures were analysed per group.
METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: without S9 mix: 4, 18 and 28 h; with S9 mix: 4 h
- Fixation time (start of exposure up to fixation or harvest of cells): without S9 mix: 18 and 28 h; with S9 mix: 18 and 28 h
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): GIEMSA
NUMBER OF CELLS EVALUATED: 100 metaphase cells per slide (200 per dose from two independent cultures)
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cell numbers - Evaluation criteria:
- A test item is classified as non-clastogenic if:
-the number of induced structural chromosome aberrations in all evaluated dose groups is in the range of our historical control data (0.0 - 4.0 % aberrant cells, exclusive gaps). and/or
-no significant increase of the number of structural chromosome aberrations is-observed.
A test item is classified as clastogenic if:
-the number of induced structural chromosome aberrations is not in the range of our historical control data (0.0 - 4.0 % aberrant cells, exclusive gaps).
and
-either a concentration-related or a significant increase of the number of structural chromosome aberrations is observed.
However, both biological and statistical significance should be considered together. If the criteria mentioned above for the test item are not clearly met, the classification with regard to the historical data and the biological relevance is discussed and/or a confirmatory experiment is performed.
Although the inclusion of the structural chromosome aberrations is the purpose of this study, it is important to include the polyploids and endoreduplications. The following criteria is valid:
A test item can be classified as aneugenic if:
-the number of induced numerical aberrations is not in the range of our historical control data (0.0 - 5.2.% polyploid cells). - Statistics:
- Statistical significance was confirmed by means of the Fisher's exact test (p < 0.05).
- 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:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no relevant effect
- Effects of osmolality: no relevant effect
- Precipitation: observed at concentrations >=2.7 µg/mL (without S9 mix, Experiment I, 4h/18h); >= 5.0 µg/ml (without S9 mix, Experiment II, 18h/18h); >= 2.5 µg/ml (without S9 mix, Experiment II, 28h/28h); >=2.7 µg/mL (with S9 mix, Experiment I, 4h/18h) and >=5.0 µg/mL (with S9 mix, Experiment II, 4h/28h)
RANGE-FINDING/SCREENING STUDIES:
No clear cytotoxicity was observed in the range-finding pre-test (test concentrations up to 170 µg/mL).
COMPARISON WITH HISTORICAL CONTROL DATA:
Results were within the range of historical control values
ADDITIONAL INFORMATION ON CYTOTOXICITY:
Cytotoxicity was not observed up to the highest concentrations tested (170 µg/mL) - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative
The test item did not induce structural chromosome aberrations in V79 Chinese Hamster lung cells in vitro without or with metabolic activation in this study. - Executive summary:
The test item (suspended in DMSO) was assessed for its potential to induce structural chromosome aberrations in V79 cells of the Chinese hamster in vitro in two independent experiments according to OECD TG 473. The highest applied concentration was determined in a pre-test on cytotoxicity and technical aspects (precipitation).
The following study design was performed with 2 independent cultures per group:
without S9 mix
with S9 mix
exp. I
exp. II
exp. I
exp. II
Exposure period
4 hrs
18 hrs
28 hrs
4 hrs
4 hrs
Preparation interval
18 hrs
18 hrs
28 hrs
18 hrs
28 hrs
The following concentrations were tested (evaluated experimental points in bold letters):
without metabolic activation (P: precipitation occured)
Experiment 1: 1.3, 2.7 (P), 5.3 (P), 10.6 (P), 21.3 (P), 42.5 (P) µg/ml
Experiment 2a: 0.3, 0.6, 1,3, 2.5, 5.0 (P), 10.0 (P) µg/ml
Experiment 2b: 0.3, 0.6, 1,3, 2.5 (P), 5.0 (P), 10.0 (P) µg/ml
with metabolic activation (P: precipitation occured)
Experiment 1: 1.3, 2.7 (P), 5.3 (P), 10.6 (P), 21.3 (P), 42.5 (P), 85.0 (P), 170 (P) µg/ml
Experiment 2: 0.3, 0.6, 1.3, 2.5, 5.0 (P), 10.0 (P) µg/ml
No toxic effects indicated by reduced mitotic indices or reduced cell numbers were observed after treatment with the test item, but precipitation occured as indicated above.
100 metaphase per culture were scored for structural chromosome aberrations. In both independent experiments, no biologically relevant increase in the number of cells carrying structural chromosomal aberrations was observed after treatment with the test item.
No relevant increase in the frequencies of polyploid metaphases was found after treatment with the test item as compared to the frequencies of the controls.
Therefore, the test item was not clastogenic in this in vitro assay.
Appropriate mutagens were used as positive controls. They induced statistically significant increases in cells with structural chromosome aberrations.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- key study
- Study period:
- From 15 OCT 2007 to 05 DEC 2007
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study (OECD 476), GLP compliant
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- HPRT
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- - Type and identity of media:supplemented MEM
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- liver S9 mix from phenobarbital/beta-naphthoflavone pretreated Wistar rats
- Test concentrations with justification for top dose:
- without S9 mix:
Experiment I: 1.3, 2.7, 5.3, 10.7, 21.3 and 170.0 µg/ml
Experiment II: 1.3, 2.7, 5.3, 10.7, 21.3 and 170.0 µg/ml
With S9 mix:
Experiment I: 1.3, 2.7, 5.3, 10.7, 21.3 and 170.0 µg/ml
Not all test groups were evaluated (for evaluation points see executive summary) - Vehicle / solvent:
- - DMSO
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- without metabolic activation
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 7,12-dimethylbenzanthracene
- Remarks:
- with metabolic activation
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: Experiment I: 4 h, experiment II: 24 h
- Expression time (cells in growth medium): 24 h
- Selection time (if incubation with a selection agent): 8 days
- Fixation time (start of exposure up to fixation or harvest of cells): 3+7+8 days
SELECTION AGENT (mutation assays): 6-thioguanine
NUMBER OF REPLICATIONS: 2 experiments, treatments performed in duplicate, both with and without metabolic activation
NUMBER OF CELLS EVALUATED: colonies with more than 50 cells
DETERMINATION OF CYTOTOXICITY
- Method: colony formation - Evaluation criteria:
- A test item is classified as positive if it induces either a concentration-related increase of the mutant frequency or a reproducible and positive response at one of the test points.
A test item producing neither a concentration- related increase of the mutant frequency nor a reproducible positive response at any of the test points is considered non-mutagenic in this system.
A positive response is described as follows:
A test item is classified as mutagenic if it reproducibly induces a mutation frequency that is three times above the spontanous mutation frequency at least at one of the concentrations in the experiment.
The test item is classified as mutagenic if there is a reproducible concentration-related increase of the mutation frequency. Such evaluation may be considered also in the case that a threefold increase of the mutant frequency is not observed.
However, in a case by case evaluation this decision depends on the level of the corresponding negative control data. If there is by chance a low spontanous mutation rate in the range normally found (0.5-31.8 mutants per 10 to the power of 6 cells) a concentration-related increase of the mutations within this range has to be discussed. The variability of the mutation rates of negative and solvent controls within all experiments of this study was also taken into considration. - Statistics:
- A linear regression was performed to assess a possible dose dependent increase of mutant frequencies.
- 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:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: from 10.7 µg/ml with and without metabolic activation in both experiments.
RANGE-FINDING/SCREENING STUDIES:
A pre-test was performed with concentrations ranging from 1.3 to 170.0 µg/ml - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative
The test substance did not increase the mutant frequency at the HPRT locus in Chinese hamster V79 cells and is therefore considered to be non-mutagenic under the conditions of the test. - Executive summary:
The test item was assessed for its potential to induce gene mutations at the HPRT locus using V79 cells of the Chinese hamster according to OECD TG 476. Chinese hamster V79 cells were treated with the test substance in the following concentrations:
without S9 mix:
Experiment I: 1.3, 2.7, 5.3, 10.7, 21.3 and 170.0 µg/ml
Experiment II: 1.3, 2.7, 5.3, 10.7, 21.3 and 170.0 µg/ml
With S9 mix:
Experiment I: 1.3, 2.7, 5.3, 10.7, 21.3 and 170.0 µg/ml
Marked in bold: cultures which were not continued.
In experiment I and IA the treatment period was 4 h with and without metabolic activation. The second experiment was performed in the absence of metabolic activation with a treatment time of 24 h.
A precipitation of the test substance was noted in both experiments at 10.7 and 170 µg/ml with and without metabolic activation. The solvent control gave mutant frequencies within the normal range. The positive controls gave marked increases in the mutant frequency, indicating the satisfactory performance of the test system. The test substance demonstrated no statistically significant or dose related increases in mutant frequency at any dose level, either with or without metabolic activation, thus indicating that the test item is not mutagenic under the conditions tested.
Referenceopen allclose all
Test Results: Experiment 1 - Without Metabolic Activation
|
|
Number of revertants (mean number of colonies per plate) |
||||
S9-Mix |
Test substance concentration (µg/plate) |
TA 100 |
TA 1535 |
TA 102 |
TA 98 |
TA 1537 |
- |
0 |
81 |
22 |
276 |
27 |
13 |
- |
25 |
70 |
25 |
226 |
12 |
9 |
- |
75 |
73 |
26 |
237 |
22 |
14 |
- |
250 |
71 |
25 |
197 |
13 |
10 |
- |
750 |
81 |
17 |
239 |
20 |
8 |
- |
2500 |
82 |
20 |
245 |
30 |
9 |
Pos. controls S9-Mix - |
Name Concentration No. colonies per plate |
ENNG |
ENNG |
MMC |
4NQO |
9AA |
3 |
5 |
0.5 |
0.2 |
80 |
||
507 |
316 |
1103 |
130 |
678 |
ENNG: N-ethyl-N'-nitro-N-nitrosoguanidine
4NQO: 4-Nitroquinoline-1-oxide
9AA: 9-Aminoacridine
MMC: Mitomycin
C
C: Contaminated
P: Precipitate
Test Results: Experiment 1 - With Metabolic Activation
|
|
Number of revertants (mean number of colonies per plate) |
||||
S9-Mix |
Test substance concentration (µg/plate) |
TA 100 |
TA 1535 |
TA 102 |
TA 98 |
TA 1537 |
+ |
0 |
86 |
19 |
209 |
23 |
17 |
+ |
25 |
88 |
21 |
218 |
20 |
13 |
+ |
75 |
82 |
19 |
215 |
23 |
10 |
+ |
250 |
87 |
22 |
210 |
21 |
9 |
+ |
750 |
90 |
18 |
231 |
26 |
15 |
+ |
2500 |
85 |
19 |
218 |
33 |
10 |
Pos. controls S9-Mix + |
Name Concentration No. colonies per plate |
2AA |
2AA |
DAN |
BP |
9AA |
1 |
2 |
10 |
5 |
2 |
||
370 |
256 |
920 |
211 |
141 |
||
CR |
|
|
CR |
|
||
50 |
|
|
50 |
|
||
593 |
|
|
221 |
|
2AA: 2-Aminoanthracene
BP: Benzo(a)pyrene
CR: Congo
Red
DAN: 1,8-Dihydroxyanthraquinone
P: Precipitate
Test Results: Experiment 2 - Without Metabolic Activation
|
|
Number of revertants (mean number of colonies per plate) |
||||
S9-Mix |
Test substance concentration (µg/plate) |
TA 100 |
TA 1535 |
TA 102 |
TA 98 |
TA 1537 |
- |
0 |
125 |
20 |
235 |
30 |
11 |
- |
25 |
107 |
19 |
227 |
34 |
9 |
- |
75 |
115 |
21 |
203 |
33 |
5 |
- |
250 |
125 |
23 |
213 |
25 |
8 |
- |
750 |
126 |
18 |
215 |
30 |
7 |
- |
2500 |
138 |
23 |
240 |
32 |
12 |
Pos. controls S9-Mix - |
Name Concentration No. colonies per plate |
ENNG |
ENNG |
MMC |
4NQO |
9AA |
3 |
5 |
0.5 |
0.2 |
80 |
||
722 |
292 |
622 |
129 |
1485 |
ENNG: N-ethyl-N'-nitro-N-nitrosoguanidine
4NQO: 4-Nitroquinoline-1-oxide
9AA: 9-Aminoacridine
MMC: Mitomycin
C
C: Contaminated
P: Precipitate
Test Results: Experiment 2 - With Metabolic Activation
|
|
Number of revertants (mean number of colonies per plate) |
||||
S9-Mix |
Test substance concentration (µg/plate) |
TA 100 |
TA 1535 |
TA 102 |
TA 98 |
TA 1537 |
+ |
0 |
128 |
14 |
258 |
23 |
12 |
+ |
25 |
126 |
11 |
262 |
27 |
8 |
+ |
75 |
127 |
14 |
246 |
22 |
13 |
+ |
250 |
134 |
17 |
254 |
31 |
13 |
+ |
750 |
138 |
12 |
248 |
23 |
15 |
+ |
2500 |
128 |
11 |
252 |
20 |
16 |
Pos. controls S9-Mix + |
Name Concentration No. colonies per plate |
2AA |
2AA |
DAN |
BP |
9AA |
1 |
2 |
10 |
5 |
2 |
||
557 |
298 |
722 |
160 |
260 |
||
CR |
|
|
CR |
|
||
50 |
|
|
50 |
|
||
468 |
|
|
194 |
|
2AA: 2-Aminoanthracene
BP: Benzo(a)pyrene
CR: Congo
Red
DAN: 1,8-Dihydroxyanthraquinone
P: Precipitate
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Gene mutation in bacteria in vitro
The ability to induce gene mutation in bacteria of the registered (target) substance was investigated in Salmonella typhimurium strain TA100 with (induced rat liver S9 mix) and without metabolic activation at concentrations of 0, 0.075, 0.25, 0.75, 2.5, 7.5, 25, 75, 250, 750 and 2500 µg/plate using the plate incorporation assay according to OECD guideline 471 and under GLP conditions (Harlan_41102439). Due to the test item’s characteristic as an azo-dye, the test was also conducted using the Prival modification, i.e. testing S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 in the preincubation assay with un-induced hamster liver S9 mix for metabolic activation. This test was performed using the following concentrations: 0, 25, 75, 250, 750 and 2500 µg/plate. The test item did not reveal any mutagenic activity under the conditions tested. The appropriate reference mutagens showed distinct positive mutagenic effects.
Chromosome aberration in vitro
The analogue substance Permanent Carmin FBB 02 (N-(4-chloro-2,5-dimethoxyphenyl)-3-hydroxy-4-[[2-methoxy-5-[(phenylamino)carbonyl]phenyl]azo]naphthalene-2-carboxamide]) suspended in DMSO was assessed for its potential to induce structural chromosome aberrations in V79 cells of the Chinese hamster in vitro in two independent experiments according to OECD TG 473 under GLP conditions (RCC_1136802). The highest applied concentration was determined in a pre-test on cytotoxicity and precipitation of the test substance. The following concentrations were tested:
without metabolic activation (P: precipitation occurred)
Experiment 1: 1.3, 2.7 (P), 5.3 (P), 10.6 (P), 21.3 (P), 42.5 (P) µg/ml
Experiment 2a: 0.3, 0.6, 1,3, 2.5, 5.0 (P), 10.0 (P) µg/ml
Experiment 2b: 0.3, 0.6, 1,3, 2.5 (P), 5.0 (P), 10.0 (P) µg/ml
with metabolic activation (P: precipitation occurred)
Experiment 1: 1.3, 2.7 (P), 5.3 (P), 10.6 (P), 21.3 (P), 42.5 (P), 85.0 (P), 170 (P) µg/ml
Experiment 2: 0.3, 0.6, 1.3, 2.5, 5.0 (P), 10.0 (P) µg/ml
No toxic effects indicated by reduced mitotic indices or reduced cell numbers were observed after treatment with the test item, but precipitation occurred as indicated above. 100 metaphases per culture were scored for structural chromosome aberrations. In both independent experiments, no biologically relevant increase in the number of cells carrying structural chromosomal aberrations was observed after treatment with the test item. No relevant increase in the frequencies of polyploid metaphases was found after treatment with the test item as compared to the frequencies of the controls. Therefore, the test item was not clastogenic in this in vitro assay. Appropriate mutagens were used as positive controls and induced statistically significant increases in cells with structural chromosome aberrations.
Gene mutation in mammalian cells in vitro
The analogue substance Permanent Carmin FBB 02 (N-(4-chloro-2,5-dimethoxyphenyl)-3-hydroxy-4-[[2-methoxy-5-[(phenylamino)carbonyl]phenyl]azo]naphthalene-2-carboxamide]) was assessed for its potential to induce gene mutations at the HPRT locus using V79 cells of the Chinese hamster according to OECD TG 476 under GLP conditions (RCC_1136801). Chinese hamster V79 cells were treated with the test substance in the following concentrations:
without S9 mix
Experiment I: 1.3, 2.7, 5.3, 10.7, 21.3 and 170.0 µg/ml
Experiment II: 1.3, 2.7, 5.3, 10.7, 21.3 and 170.0 µg/ml
With S9 mix
Experiment I: 1.3, 2.7, 5.3, 10.7, 21.3 and 170.0 µg/ml
In experiment I the treatment period was 4 h with and without metabolic activation. The second experiment was performed in the absence of metabolic activation with a treatment time of 24 h. A precipitation of the test substance was noted in both experiments at 10.7 and 170 µg/ml with and without metabolic activation. The solvent control gave mutant frequencies within the normal range. The positive controls gave marked increases in the mutant frequency, indicating the satisfactory performance of the test system. The test substance demonstrated no statistically significant or dose related increases in mutant frequency at any dose level, either with or without metabolic activation, thus indicating that the test item is not mutagenic under the conditions tested.
Justification for classification or non-classification
The available data on genetic toxicity in vitro obtained with the registered substance and an analogue substance do not meet the criteria for classification according to Regulation (EC) No. 1272/2008 (CLP) and are therefore conclusive but not sufficient for classification.
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.