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EC number: 228-768-4 | CAS number: 6358-31-2
- 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
Pigment Yellow 74 also yielded negative results in an in vitro gene mutation study in mammalian cells (mouse lymphoma assay) in concentration up to 2000 µg/ml in the presence and absence of metabolic activation. This study was selected as key study.
Pigment Yellow 74 did not induce micronuclei in V79 cells (Chinese hamster cell line) in vitro in the absence and the presence of metabolic activation under the experimental conditions used when test up to precipitating concentrations.
In conclusion, Pigment Yellow 74 is not mutagenic in the bacterial reverse mutation assay, in the mouse lymphoma assay and in the in vitro micronucleus assay in the presence and absence of metabolic activation up to the tested concentrations.
Mutagenic properties of Pigment Yellow 74 were investigated in
several bacterial reverse mutation assays (Ames test; test strains used:
S. typhimurium TA 97, TA 98, TA 100, TA 1535, TA 1537, TA 1538, E. coli
WP2 uvr A), in an in vitro gene mutation study in mammalian cells (Mouse
lymphoma assay) and in an in vitro micronucleus assay. Negative results
were obtained in all tests with and without metabolic activation.
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:
- 29 November 2006 to 08 December 2006
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: performed in accordance with OECD and GLP guidelines
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- according to German Chemikaliengesetz and OECD Principles of Good Laboratory Practice
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- E. coli WP2 uvr A
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- Pre-Experiment and Experiment I: 0 (control), 3, 10, 33, 100, 333, 1000, 2500, 5000 µg/plate
Experiment II: 0 (control), 33, 100, 333, 1000, 2500, 5000 µg/plate - Vehicle / solvent:
- Ethanol
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Ethanol
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: see below for additional information
- Details on test system and experimental conditions:
- The assay was performed in two independent experiments:
experiment I: plate incorporation assay with and without induced rat liver S9 mix
experiment II: pre-incubation test with and without non-induced hamster liver S9 mix
Hamster liver S9 mix, but not rat liver S9 mix, contained the reductive agent FMN.
DURATION
- Preincubation period: 30 min, 30°C
- Exposure duration: after solidification the plates were incubated upside down for at least 48 hours at 37°C in the dark
NUMBER OF REPLICATIONS: 3
DETERMINATION OF CYTOTOXICITY: existence of evaluable plates (> 0 colonies) at five concentrations or more
POSITIVE CONTROL SUBSTANCES:
without metabolic activation: sodium azide (TA 1535, TA 100), 4-nitro-o-phenylene-diamine ((TA 1537, TA 98), methyl methane sulfonate (WP2 uvrA);
with metabolic activation: 2-aminoanthracene (all strains with rat liver S9 mix and TA 1535, TA 100, TA 1537, WP2 uvrA with hamster liver S9 mix), cKongo red (TA 98 with hamster liver S9 mix) - Evaluation criteria:
- A test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA 98, TA 100, and WP2 uvrA) or thrice (strains TA 1535 and TA 1537) the colony count of the corresponding solvent control is observed.
A dose dependent increase is considered biologically relevant if the threshold is exceeded at more than one concentration.
An increase exceeding the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment.
A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls such an increase is not considered biologically relevant. - Key result
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Experiment I, TA 1537, without metabolic activation: minor reduction in number of revertants at 5000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Interpretation of results:
negative with and without metabolic activation
During the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by
frameshifts or base-pair substitutions in the genome of the strains used. Therefore, the test item is considered to be non-mutagenic in this Salmonella typhimurium and Escherichia coli reverse mutation assay. - Executive summary:
This study was performed to investigate the potential of the test substance to induce gene mutations according to the plate incorporation assay with rat liver S9 (experiment I) and the pre-incubation test with hamster liver S9 (experiment II) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100, and the Escherichia coli strain WP2 uvrA.
The assay was performed in two independent experiments with and without liver microsomal activation. Each concentration, including the controls, was tested in triplicate. The test item was tested at the following concentrations:
Pre-Experiment/Experiment I : 3; 10; 33; 100; 333; 1000; 2500; and 5000 µg/plate
Experiment II: 33; 100; 333; 1000; 2500; and 5000 µg/plate
The plates incubated with the test item showed normal background growth up to 5000 µg/plate with and without metabolic activation in both experiments.
No toxic effects, evident as a reduction in the number of revertants, were observed with and without metabolic activation in all strains. Only in experiment I in strain TA 1537 in the absence of metabolic activation a minor reduction in the number of revertants, were observed at 5000 µg/plate.
No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with the test item at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.
Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies.
- Endpoint:
- in vitro cytogenicity / micronucleus study
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: According to Draft Proposal for a new Guideline No. 487
- Qualifier:
- according to guideline
- Principles of method if other than guideline:
- OECD Guideline Draft Proposal for a new Guideline No. 487, Version 3
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian cell micronucleus test
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- Thawed stock cultures were propagated at 37 °C in 80 cm2 plastic flasks (Greiner,
72632 Frickenhausen, Germany). About 5 x 105 cells per flask were seeded in 15 mL of MEM (minimal essential medium; Invitrogen GIBCO, 76131 Karlsruhe, Germany) supplemented with 10 % fetal calf serum (FCS; Invitrogen, 76131 Karrlsruhe, Germany). Additionally, the medium was supplemented with 1 % 100x Penicillin/ Streptomycin solution (10.000 Units/mL Penicillin, 10 mg/mL Streptomycin; PAA Laboratories GmbH, 35091 Cölbe, Germany) and 1 % Amphotericin B (250 µg/mL, PAA Laboratories GmbH, 35091 Cölbe, Germany). The cells were subcultured twice weekly. The cell cultures were incubated at 37 °C in a humidified atmosphere with 4.5 % carbon dioxide (95.5% air). - Metabolic activation:
- with and without
- Metabolic activation system:
- rat liver S9
- Test concentrations with justification for top dose:
- Exp. I: with and without S9 mix: 4.7, 9.4, 18.8, 37.5, 75.0, 150.0, 300.0, 600.0, and 1200.0 µg/mL
Exp. II: without S9 mix: 2.4, 4.7, 9.4, 18.8, 37.5, 75.0, 150.0, 300.0, and 600.0 µg/mL
with S9 mix: 1.2, 2.4, 4.7, 9.4, 18.8, 37.5, 75.0, and 150.0 µg/mL. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Ethanol (EtOH) (E. MERCK, 64293 Darmstadt, Germany; purity 99.8 %)
- Justification for choice of solvent/vehicle: The solvent was chosen to its solubility properties and its relative non-toxicity to the cell cultures. - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- EtOH
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- griseofulvin, cyclophosphamide
- Details on test system and experimental conditions:
- Two independent experiments were performed. In Experiment I the exposure period was 4 hours with and without metabolic activation. In Experiment II the exposure period was 24 hours without S9 mix and 4 hours with metabolic activation. The cells were prepared 24 hours after start of treatment with the test item.
METHOD OF APPLICATION: in minimal essential medium
DURATION
- Exposure duration: 4 and 24 hours
- Expression time (cells in growth medium): 24 hours
- Fixation time (start of exposure up to fixation or harvest of cells): 24 hours
SPINDLE INHIBITOR (cytogenetic assays):
STAIN (for cytogenetic assays): May Gruenwald and Giemsa
NUMBER OF REPLICATIONS: 1.5 - 2
NUMBER OF CELLS EVALUATED: 2000
EVALUATION: Evaluation of the cultures was performed manually using NIKON microscopes with 40 x oil immersion objectives. The micronuclei were counted in cells showing a clearly visible cytoplasm area. The criteria for the evaluation of micronuclei are described in the publication of Countryman and Heddle (1976). The micronuclei were stained in the same way as the main nucleus. The area of the micronucleus did not extend the third part of the area of the main nucleus. 2000 cells from clones with 2 - 8 cells were scored per test group. The frequency of micronucleated cells was reported as % micronucleated cells.
DETERMINATION OF CYTOTOXICITY
- Method: Proliferation Index
OTHER EXAMINATIONS:
- Evaluation criteria:
- A test item can be classified as mutagenic if:
- the number of micronucleated cells is not in the range of the historical control data, and
- either a concentration-related increase in three test groups or a significant increase of micronucleated cells in at least one test group is observed.
A test item can be classified as non-mutagenic if:
- the number of micronucleated cells in all evaluated test groups is in the range of the historical control data, and/or
- no concentration-related increase in the number of micronucleated cells is observed.
Statistical significance can be confirmed by means of the Chi square test. However, both biological and statistical significance should be considered together. If the criteria above mentioned 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. - Statistics:
- Statistical significance can be confirmed by means of the Chi square test.
- 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
- Positive controls validity:
- valid
- Additional information on results:
- The test item, suspended in ethanol, was assessed for its potential to induce micronuclei in Chinese hamster V79 cells in vitro in the absence and the presence of metabolic activation by S9 mix.
Two independent experiments were performed. In Experiment I the exposure period was 4 hours with and without metabolic activation. In Experiment II the exposure period was 24 hours without S9 mix and 4 hours with metabolic activation. The cells were prepared 24 hours after start of treatment with the test item.
In each experimental group two parallel cultures were analysed and 1000 cells per culture were scored for micronuclei.
In both experiments no cytotoxicity was observed up to the highest evaluated concentration. However, due to strong test item precipitation higher concentrations were not evaluable for cytogenetic damage. In addition, no relevant influence of the test item on the pH value or osmolarity was observed (Exp. I: solvent control 377 mOsm, pH 7.6 versus 344 mOsm and pH 7.6 at 1200 µg/mL; Exp. II: solvent control 381 mOsm, pH 7.6 versus 373 mOsm and pH 7.5 at 1200 µg/mL).
In Experiment I in the absence of metabolic activation no statistically significant and biologically relevant increase in the number of micronucleated cells was observed at the evaluated concentrations. In the presence of metabolic activation one increase in micronucleated cells (2.10 %) was observed after treatment with 4.7 µg/mL . This value slightly exceeded the laboratory´s historical control data range (0.15 – 1.70 % micronucleated cells), but was dose-independently and statistically not significant.
In Experiment II in the absence and presence of metabolic activation no statistically significant and biologically relevant increase in micronucleated cells was observed at the evaluated concentrations. The slight increase in the number of micronucleated cells obtained in Experiment I in the presence of metabolic activation was not confirmed. Therefore, this observation is regarded as biologically irrelevant.
Griseofulvin (9.0 µg/mL), Mitomycin C (0.03 and 0.1 µg/mL), and CPA (10 and 15 µg/mL) were evaluated as positive controls and showed a distinct increase in the percentage of micronucleated cells.
In conclusion, it can be stated that under the experimental conditions reported, the test item did not induce micronuclei in V79 cells (Chinese hamster cell line) in vitro in the absence and the presence of metabolic activation. - Conclusions:
- Interpretation of results:
negative with and without metabolic activation
The test item did not induce micronuclei in V79 cells (Chinese hamster cell line) in vitro in the absence and the presence of metabolic activation under the experimental conditions used. Therefore, the test item has to be considered as non-mutagenic in this in vitro test system when tested up to precipitating or the highest evaluated test item concentrations. - Executive summary:
The test item, suspended in ethanol, was assessed for its potential to induce micronuclei in Chinese hamster V79cells in vitro in the absence and the presence of metabolic activation by S9 mix.
In each experimental group two parallel cultures were analysed and 1000 cells per culture were scored for micronuclei.
The following test item concentrations were applied (e=evaluated):
Exp. I: with and without S9 mix: 4.7 (e), 9.4 (e), 18.8 (e), 37.5, 75.0, 150.0, 300.0, 600.0, and 1200.0 µg/mL
Exp. II: without S9 mix: 2.4 (e), 4.7 (e), 9.4 (e), 18.8, 37.5, 75.0, 150.0, 300.0, and 600.0 µg/mL
Exp. II: with S9 mix: 1.2 (e), 2.4 (e), 4.7 (e), 9.4, 18.8, 37.5, 75.0, and 150.0 µg/mL.
The highest applied concentration (1200 µg/mL; approx. 3 mM) was chosen with regard to the solubility properties of the test item in ethanol following the current Draft Proposal for a new Guideline No. 487. Test item precipitation was observed at 37.5 µg/mL and higher.
In the absence and the presence of S9 mix, no cytotoxicity was observed up to the highest evaluated concentration. Higher concentrations were not evaluable for cytogenetic damage because strong test item precipitation occurred in any step of evaluation.
In Experiment I no clastogenicity was observed at the concentrations evaluated in the absence of S9 mix. In the presence of S9 mix a dose-independent and statistically not significant induction of micronucleated cells (2.10 %) was observed after treatment with 4.7 µg/mL. This value marginally exceeded the laboratory’s historical control data range (0.15 – 1.70 % micronucleated cells).
In Experiment II in the absence and the presence of S9 mix no clastogenicity was observed at the concentrations evaluated. The slight increase in micronucleated cells obtained in Experiment I in the presence of S9 mix was not confirmed. Therefore, this observation has to be regarded as biologically irrelevant.
Appropriate mutagens were used as positive controls. They induced statistically significant increases (p<0.05) in the percentage of micronucleated cells.
In conclusion, it can be stated that under the experimental conditions reported, the test item did not induce micronuclei in V79 cells (Chinese hamster cell line) in vitro in the absence and the presence of metabolic activation. Therefore, the test item has to be considered as non-mutagenic in this in vitro test system when tested up to precipitating or the highest evaluated test item concentrations.
- 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:
- year of publication: 1987
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- It is unclear from the report whether the test was performed according to OECD and GLP guidelines. Important aspects (duplicate cultures, dosing range) in line with current OECD guideline, but study design is restricted because no second experiment with 24-hour incubation was done.
- Reason / purpose for cross-reference:
- reference to same study
- Principles of method if other than guideline:
- L5178Y TK +/- mouse lymphoma assay
- GLP compliance:
- not specified
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- TK
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Fischer's medium for leukemic cells of mice (supplemented with 10% horse serum)
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: no data
- Periodically "cleansed" against high spontaneous background: no data - Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- 0 (control), 972.0, 1231.0, 1488.0, 1744.0, 2000.0 µg/ml
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO or acetone
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- see below for additional information
- Details on test system and experimental conditions:
- Number of Replications: 2
- positve control substances:
without metabolic activation: ethyl methanesulfonate
with metabolic activation: 3-methylcholanthrene
DURATION
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 48 hours
- Selection time (if incubation with a selection agent): 12 days
SELECTION AGENT (mutation assays): trifluorothymidine - Evaluation criteria:
- A response was considered positive if there was a dose-related increase in the mutant frequency above the spontaneous control frequency, with a 2-fold increase at more than 1 dose and relative total growth greater than 10%.
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Conclusions:
- Interpretation of results:
negative with and without metabolic activation
Pigment Yellow 74 was not mutagenic under the conditions tested. - Executive summary:
Pigment Yellow 74 was tested in a mammalian cell gene mutation assay (mouse lymphoma L5178Y cells) in the presence and absence of metabolic activation. Relative cloning efficiency and growth as well as mutant frequency were not affected by the test substance (concentration range tested: 972.0 to 2000.0 µg/ml). Pigment Yellow 74 was not mutagenic under the conditions tested.
Referenceopen allclose all
Mean mutant number ratios treated/solvent control
Exp. I: plate incorporation method without S9 mix
Concentrations given in µg/plate
Strain -- 3 -- 10 -- 33 -- 100 -- 333 -- 1000 -- 2500 -- 5000
TA1535 -- 1.0 -- 0.8 -- 0.8 -- 1.1 -- 0.8 -- 0.7 -- 0.9 -- 0.7
TA1537 -- 1.5 -- 1.1 -- 0.9 -- 0.9 -- 0.9 -- 1.3 -- 0.9 -- 0.4
TA98 -- 0.9 -- 1.0 -- 1.0 -- 1.1 -- 0.8 -- 0.9 -- 0.9 -- 0.7
TA100 -- 0.8 -- 0.9 --1.0 -- 1.1 -- 0.9 -- 1.0 -- 0.9 -- 0.8
WP2uvrA -- 1.0 -- 1.0 -- 1.1 -- 1.2 -- 0.9 -- 0.9 -- 0.8 -- 0.8
Exp. I: plate incorporation method with rat S9 mix
Concentrations given in µg/plate
Strain -- 3 -- 10 -- 33 -- 100 -- 333 -- 1000 -- 2500 -- 5000
TA1535 -- 1.1 -- 1.1 -- 1.1 -- 1.0 -- 1.1 -- 1.2 -- 1.2 -- 0.7
TA1537 -- 1.1 -- 0.9 -- 1.1 -- 1.3 -- 1.1 -- 0.9 -- 0.9 -- 0.7
TA98 -- 1.1 -- 1.1 -- 1.1 -- 1.0 -- 0.9 -- 0.9 -- 1.0 -- 0.8
TA100 -- 1.0 -- 1.1 --1.0 -- 1.0 -- 1.0 -- 1.0 -- 0.9 -- 0.8
WP2uvrA -- 1.0 -- 1.0 -- 0.9 -- 0.9 -- 0.9 -- 0.8 -- 0.8 -- 0.8
Exp. II: pre-incubation method without S9 mix
Concentrations given in µg/plate
Strain -- 33 -- 100 -- 333 -- 1000 -- 2500 -- 5000
TA1535 -- 1.2 -- 0.9 -- 1.0 -- 0.9 -- 1.0 -- 0.9
TA1537 -- 1.2 -- 1.3 -- 1.4 -- 1.4 -- 1.1 -- 0.9
TA98 -- 1.0 -- 1.2 -- 0.9 -- 1.1 -- 0.8 -- 0.7
TA100 --1.2 -- 1.1 -- 1.1 -- 1.1 -- 0.9 -- 1.0
WP2uvrA -- 1.2 -- 0.9 -- 1.1 -- 0.9 -- 1.0 -- 0.9
Exp. II: pre-incubation method with hamster S9 mix
Concentrations given in µg/plate
Strain -- 33 -- 100 -- 333 -- 1000 -- 2500 -- 5000
TA1535 -- 0.9 -- 0.8 -- 0.7 -- 0.7 -- 0.7 -- 0.7
TA1537 -- 1.3 -- 1.0 -- 1.1 -- 0.8 -- 0.8 -- 0.8
TA98 -- 1.0 -- 1.0 -- 1.0 -- 1.0 -- 0.7 -- 0.7
TA100 --1.0 -- 0.8 -- 1.0 -- 0.9 -- 0.8 -- 0.7
WP2uvrA -- 1.2 -- 0.8 -- 0.7 -- 1.0 -- 0.9 -- 0.7
Dose selection was performed following the current Draft Proposal for a new Guideline No. 487. In general, the test item should be tested up to a maximum concentration of 5 mg/mL, 5 µL/mL, or 10 mM if the test item is not toxic. The highest treatment concentration chosen for the evaluation of genotoxicity should reduce the cell growth to approx. 50 %, determined by a Proliferation Index (PI). The solubility of the test item and changes in the pH value and the osmolarity may influence the dose selection.
With respect to the solubility of the test item, a concentration of 1200 µg/mL of the test item (approx. 3 mM) was applied as top concentration for treatment of the cultures in Experiment I. Dose selection of Experiment II was influenced by the results obtained in Experiment I. In Experiment I precipitation of the test item in culture medium occurred at 37.5 µg/mL in the absence and presence of metabolic activation. In Experiment II concentrations between 2.4 and 600 µg/mL in the absence of metabolic activation as well as concentrations between 1.2 and 150 µg/mL in the presence of metabolic activation were applied.
Summary of results of the micronucleus test
Exp. |
Preparation |
Test item |
Proliferation |
Micronucleated |
interval |
concentration |
Index |
cells |
|
in µg/mL |
in % |
|||
Exposure period 4 hrs without S9 mix |
||||
I |
24 hrs |
Solvent control1 |
2.68 |
0.55 |
Positive control2 |
2.57 |
10.85S |
||
9.4 |
2.63 |
0.90 |
||
18.8 |
2.58 |
1.05 |
||
37.5P |
2.80 |
0.50 |
||
Exposure period 24 hrs without S9 mix |
||||
II |
24 hrs |
Solvent control1 |
2.70 |
0.40 |
Positive control3 |
2.11 |
13.40S |
||
Positive control4 |
2.35 |
7.15S |
||
2.4 |
2.69 |
0.40 |
||
4.7 |
2.74 |
0.25 |
||
9.4 |
2.75 |
0.20 |
||
Exposure period 4 hrs with S9 mix |
||||
I |
24 hrs |
Solvent control1 |
1.95 |
1.35 |
Positive control5 |
1.71 |
19.05S |
||
4.7 |
1.84 |
2.10 |
||
9.4 |
1.96 |
1.50 |
||
18.8 |
2.00 |
1.05 |
||
Exposure period 4 hrs with S9 mix |
||||
II |
24 hrs |
Solvent control1 |
2.28 |
0.85 |
Positive control6 |
1.72 |
10.40S |
||
1.2 |
1.99 |
0.90 |
||
2.4 |
2.21 |
1.10 |
||
4.7 |
2.23 |
0.65 |
S Number
of micronucleated cells statistically significant higher than
corresponding
control values
P Precipitate
1 EtOH 0.5 % (v/v)
2 Mitomycin C 0.03 µg/mL
3 Mitomycin C 0.1 µg/mL
4 Griseofulvin 9.0 µg/mL
5 CPA 10.0 µg/mL
6 CPA 15.0 µg/mL
Results without rat liver S9 mix:
Concentrations (µg/ml): 0 -- 972 -- 972 -- 1231 -- 1231 -- 1488 -- 1488 -- 1744 -- 1744 -- 2000 -- 2000
Rel. suspension growth (%): x -- 75 -- 86 -- 100 -- 93 -- 94 -- 85 -- 86 -- 54 -- 53 -- 74
Rel. cloning efficiency (%): x -- 110 -- 118 -- 92 -- 110 -- 97 -- 101 -- 108 -- 103 -- 99 -- 109
Rel. total growth (%): x -- 82 -- 102 -- 92 -- 103 -- 91 -- 85 -- 93 -- 55 -- 52 -- 80
Average number of colonies trifluorothymidine / viable: x -- 73/178 -- 53/191 -- 42/149 -- 64/179 -- 58/157 -- 70/163 -- 50/175 -- 62/167 -- 54/160 -- 85 -176
Mutant frequency (per 10E4 cells): 0.77 -- 0.82 -- 0.55 -- 0.56 -- 0.72 -- 0.74 -- 0.86 -- 0.57 -- 0.74 --0.68 -- 0.97
Results with rat liver S9 mix:
Concentrations (µg/ml): 0 -- 972 -- 972 -- 1231 -- 1231 -- 1488 -- 1488 -- 1744 -- 1744 -- 2000 -- 2000
Rel. suspension growth (%): x -- 110 -- 115 -- 109 -- 114 -- 105 -- 103 -- 108 -- 104 -- 110 -- 81
Rel. cloning efficiency (%): x -- 113 -- 119 -- 109 -- 106 -- 113 -- 99 -- 104 -- 121 -- 109 -- 99
Rel. total growth (%): x -- 124 -- 137 -- 119 -- 121 -- 118 -- 102 -- 113 -- 126 -- 120 -- 80
Average number of colonies trifluorothymidine / viable: x -- 69/181 -- 65/191 -- 74/175 -- 77/169 -- 59/180 -- 61/158 -- 73/166 -- 79/194 -- 77/174 -- 43/159
Mutant frequency (per 10E4 cells): 0.76 -- 0.68 -- 0.85 -- 0.56 -- 0.91 -- 0.66 -- 0.77 -- 0.88 -- 0.81 --0.8 -- 0.54
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Justification for classification or non-classification
Pigment Yellow 74 does not have to be classified for mutagenicity according to the criteria laid down in the EU Dangerous Substances Directive (67/548/EEC) and in the EU Classification Labelling and Packaging Regulation (1272/2008/EC) because Pigment Yellow 74 did not reveal any mutagenic effect in the bacterial reverse mutation assay in the presence or absence of metabolic activation at concentrations up to 10000 µg/plate, in the mouse lymphoma assay at concentrations up to 2000 µg/ml and in the in vitro micronucleus test in V79 cells at up to a precipitating concentration of 37.5 µg/ml.
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