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EC number: 242-355-6 | CAS number: 18472-87-2
- 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
![](https://echa.europa.eu./o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
The Ames Salmonella/mammalian-microsome assay was used to evaluate the bacterial mutagenicity of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) (CAS Number: 18472-87-2) .3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) did not showed any mutagenic activity at concentrations up to 100 μg/plate in S.typhimurium strains TA1535, TA100, TA1537, TA1538 and TA98 in the presence and absence of S9 metabolic activation system and hence, according to CLP criteria, it can be concluded that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) is non genotoxic.
Chromosomal Aberration study:
C.I Pigment red 23 (30 to microgram / ml) was found to be negative for induction of chromosomal aberrations in Chinese hamster ovary cells, with and without metabolic activation.
Gene Mutation Studies in Mammalian cells
Erythrosine failed to induce gene mutation in Bacillus subtilis H17 (Rec+) and M45 (Rec-) in the rec assay.
Under the test conditions the test item 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid red 92) (CAS No. 18472-87-2) was considered to be non mutagenic in this assay.
Link to relevant study records
- Endpoint:
- in vitro DNA damage and/or repair study
- Remarks:
- Type of genotoxicity: DNA damage and/or repair
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Data is from peer-reviewed scientific journal
- Principles of method if other than guideline:
- Genetic toxicity were performed for erythrosine in Bacillus subtilis/: H17(rec+)and M45(Rec-) using Bacillus subtilis recombination assay.
- GLP compliance:
- not specified
- Type of assay:
- Bacillus subtilis recombination assay
- Species / strain / cell type:
- other: Bacillus subtilis/: H17(rec+)and M45(Rec-)
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- not specified
- Test concentrations with justification for top dose:
- 10µM
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used:DMSO
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- not specified
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: as impregnation on paper disk
DURATION
- Exposure duration: 24 hrs
- Expression time (cells in growth medium): 24 hrs - Evaluation criteria:
- The length of growth inhibition of the bacterial streak was measured
- Statistics:
- No data available
- Species / strain:
- other: Bacillus subtilis/: H17(rec+)and M45(Rec-)
- Metabolic activation:
- not specified
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative
Erythrosine failed to induce gene mutation in Bacillus subtilis H17 (Rec+) and M45 (Rec-) in the rec assay. - Executive summary:
Erythrosine was tested for gene mutation in vitro in the bacterium Basillus subtilis H17 (Rec+) and M45 (Rec-) in the rec assay performed.
Agar plates were streaked with inocula of two strains and paper disk 8 mm of diameter, immersed with 10 µM of dye in DMSO solution, was placed on the surface so as to cover the beginning of the bacterial streaks. After incubation for 24 hrs at 37◦C, the length of growth inhibition of the bacterial streak was measured.
The length of growth inhibition was found to be 2mm in Rec+and 3mm in Rec-strains.
Erythrosine failed to induce gene mutation in Bacillus subtilis H17 (Rec+) and M45 (Rec-) in the rec assay.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- secondary literature
- Justification for type of information:
- Data is from secondary source
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- GLP compliance:
- not specified
- Type of assay:
- mammalian cell gene mutation assay
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): Fuchsia Red
Molecular formula: C20H2Br4Cl4O5Na2
- Molecular weight: 829.66 g/mol
- Substance type: Organic
- Physical state: Red powder
- Purity: 99.5 area % (HPLC), 85 % (w/w) (NMR, calculated as sodium salt). - Target gene:
- TK locus
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital/Naphthoflavone treated rat liver homogenate (S 9)
- Test concentrations with justification for top dose:
- Exp.1: -S9: 5, 10, 20, 30, 40 μg/ml
+S9: 5, 10, 20, 40, 60 μg/ml
Exp.2: -S9: 19.5, 39, 78, 156 μg/ml - Vehicle / solvent:
- No data available.
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 3-methylcholanthrene
- methylmethanesulfonate
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration:
Exp.1: (-S9): 4 hours
(+S9): 4 hours
Exp.2: -(S9): 24 hours
NUMBER OF REPLICATIONS: Yes, two - Evaluation criteria:
- No data available.
- Statistics:
- No data available.
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- Toxicity: in a preliminary experiment a relevant toxicity was observed at 39.1 μg/ml in all conditions at 4 h of treatment; in the 24 h of treatment a severe toxicity was observed at 312.5 μg/ml. The doses for the main experiments were chosen on the base of a preliminary toxicity experiment.
Mutagenicity:
A) positive controls
MMS (-S9): 4 h: 1st culture: small colonies: 164 (51); large colonies:114 (51); 2nd culture: small colonies: 225(53),large colonies:157 (49).(in brackets the untreated control values)
24 h: small colonies: 1247 (61); large colonies: 346 (43)
3-MC (+S9):4 h.1st culture: small colonies: 260 (67); large colonies: 311 (67);
2nd culture: small colonies: 137 (44); large colonies: 121 (40).
The results obtained with the positive controls make acceptable the study.
B) Treated cells
A mutagenic effect observed in one culture was not repeated in the replicate culture. - Conclusions:
- Under the test conditions the test item 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid red 92) (CAS No. 18472-87-2) was considered to be non mutagenic in this assay.
- Executive summary:
In vitro Mammalian Cell Gene Mutation assay was performed to evaluate the mutagenic potential of the test compound 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid red 92) (CAS No. 18472-87-2) in the Mouse lymphoma L5178 cells (TK locus).
Two experiments were performed: at different concentration and for different time duration
Exp.1:-S9: 5, 10, 20, 30, 40 μg/ml (4 hours)
+S9: 5, 10, 20, 40, 60 μg/ml (4 hours)
Exp.2:-S9: 19.5, 39, 78, 156 μg/ml (24 hours).
In a preliminary experiment a relevant toxicity was observed at 39.1 μg/ml in all conditions at 4 h of treatment; in the 24 h of treatment a severe toxicity was observed at 312.5 μg/ml. The doses for the main experiments were chosen on the base of a preliminary toxicity experiment. In treated cell a mutagenic effect observed in one culture was not repeated in the replicate culture. Therefore under the test conditions tested the test item 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid red 92) (CAS No. 18472-87-2) was considered to be non mutagenic in this assay.
In accordance with the CLP classification, the test material does not classify as a gene mutant.
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Justification for type of information:
- Data is from peer-reviewed journal.
- Qualifier:
- no guideline available
- Principles of method if other than guideline:
- The Ames Salmonella/mammalian-microsome assay was used to evaluate the bacterial mutagenicity of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo- 6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) (CAS Number: 18472-87-2) .
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): Phloxine B
- Molecular formula: C20H2Br4Cl4O5Na2
- Molecular weight: 829.66 g/mol
- Substance type: Organic
- Physical state: Red powder
- Purity: 7-9% - Target gene:
- Histidine operon
- Species / strain / cell type:
- S. typhimurium, other: TA1535, TA100, TA1537, TA1538, TA98
- Metabolic activation:
- with and without
- Metabolic activation system:
- Liver-microsome preparation from rats injected with Aroclor 1254, 0.25 ml "S-9 mix"-plate with 0.2 ml S9/ml 'S9 mix'.
- Test concentrations with justification for top dose:
- 10,50 and 100 µg/plate
- Vehicle / solvent:
- DMSO
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- ethylmethanesulphonate
- methylmethanesulfonate
- other:
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar
DURATION
- Preincubation period: No data available
- Exposure duration:3 days - Evaluation criteria:
- Mutations in Salmonella typhimurium i.e. Number of His + Revertants/plate
- Statistics:
- Rounded mean ± standard deviation
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other:
- Remarks:
- slight toxicity was observed at top dose
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- valid
- 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:
- other:
- Remarks:
- slight toxicity was observed at top dose
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other:
- Remarks:
- slight toxicity was observed at top dose
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other:
- Remarks:
- slight toxicity was observed at top dose
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other:
- Remarks:
- slight toxicity was observed at top dose
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- COMPARISON WITH HISTORICAL CONTROL DATA: Yes, historical negative control were used.
- Remarks on result:
- other: no mutagenic potential
- Conclusions:
- The substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxo xanthen-9-yl)benzoic acid (Phloxine B) did not showed any mutagenic activity at concentrations up to 100 μg/plate in S.typhimurium strains TA1535, TA100, TA1537, TA1538 and TA98 in the presence and absence of S9 metabolic activation system and hence, it can be concluded that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) is non genotoxic.
- Executive summary:
The Ames Salmonella/mammalian-microsome assay was used to evaluate the bacterial mutagenicity of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) (CAS Number: 18472-87-2) .3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) did not showed any mutagenic activity at concentrations up to 100 μg/plate in S.typhimurium strains TA1535, TA100, TA1537, TA1538 and TA98 in the presence and absence of S9 metabolic activation system and hence, according to CLP criteria, it can be concluded that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) is non genotoxic.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Remarks:
- Read across based on category approach
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Justification for type of information:
- The study contains experimental data of a read-across analogue
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- The study was conducted to assess the mutagenicity of C.I. Pigment Red 23. To access mutagenicity, chromosomal aberration test was conducted.
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian chromosome aberration test
- Specific details on test material used for the study:
- Name- C.I Pigment Red
Physical form- Bluish red powder
Purity- > 96%
Solubility- It is insoluble in water, poorly soluble in ethanol or xylene, and highly soluble in 5% sodium carbonate solution or in oleic acid - Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix (metabolic activation enzymes and cofactors from Aroclor 1254-induced male Sprague-Dawley rat or Syrian hamster liver)
- Test concentrations with justification for top dose:
- 100 microgram/ml
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- no
- Details on test system and experimental conditions:
- In the Abs test without S9, cells were incubated in McCoy's 5A medium with C.1. Pigment Red 23 for 10 hours; Colcemid was added and incubation continued for 2 to 3 hours. The cells were then harvested by mitotic shake-off, fixed, and stained with Giemsa. For the Abs test with S9, cells were treated with C.1. Pigment Red 23 and S9 for 2 hours, after which the treatment medium was removed and the cells incubated for 11 hours in fresh medium, with Colcemid present for the final 2 hours. Cells were harvested in the same manner as for the treatment without S9.
The harvest time for the Abs test was based on the cell cycle information obtained in the
SCE test; if cell cycle delay was anticipated, the incubation period was extended. - Evaluation criteria:
- Cells were selected for scoring on the basis of good morphology and completeness of karyotype
(21 ± 2 chromosomes). All slides were scored blind and those from a single test were read by the same person. For the SCE test, usually 50 second-division metaphase cells were scored for frequency of SCEs per cell from each dose level; 200 first-division metaphase cells were scored at each dose level for the Abs test. Classes of aberrations included simple (breaks and terminal deletions), complex (rearrangements and translocations), and other (pulverized cells, despiralized chromosomes, and cells containing 10 or more aberrations). - Statistics:
- Abs data are presented as percentage of cells with aberrations. As with the SCE assay, both the dose response curve and individual dose points were statistically analyzed. A statistically significant (P
- Key result
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not specified
- Remarks on result:
- other: Non Mutagenic
- Conclusions:
- C.I Pigment red 23 (30 to microgram / ml) was found to be negative for induction of chromosomal aberrations in Chinese hamster ovary cells, with and without metabolic activation.
- Executive summary:
The study was conducted to access the mutagenicity of C.1. Pigment Red 23. Chromosomal aberration was assessed in Chinese hamster ovary cells, with and without metabolic activation. In the Abs test without S9, cells were incubated in McCoy's 5A medium with C.1. Pigment Red 23 for 10 hours; Colcemid was added, and incubation continued for 2 to 3 hours. The cells were then harvested by mitotic shake-off, fixed, and stained with Giemsa. For the Abs test with S9, cells were treated with C.1. Pigment Red 23 and S9 for 2 hours, after which the treatment medium was removed and the cells incubated for 11 hours in fresh medium, with Colcemid present for the final 2 hours. Cells were harvested in the same manner as for the treatment without S9. The harvest time for the Abs test was based on the cell cycle information obtained in the SCE test; if cell cycle delay was anticipated, the incubation period was extended.
Statistical analyses were conducted on both the slopes of the dose-response curves and the individual dose points. Abs data are presented as percentage of cells with aberrations. As with the SCE assay, both the dose response curve and individual dose points were statistically analyzed. A statistically significant (P < O.05) difference for one dose point was considered weak evidence for a positive response; significant differences for two or more doses indicated the trial was positive.
C.I Pigment red 23 was found to be negative for induction of chromosomal aberrations in Chinese hamster ovary cells, with and without metabolic activation.
Referenceopen allclose all
Table: Mutagenicity Testing Of D And C Red No. 27 With The Salmonella/Microsome Test
Chemical |
Dose |
Microsome activation |
Number of His+Revertants/plate |
||||
TA1535 |
TA100 |
TA1537 |
TA1538 |
TA98 |
|||
Negative controls |
|||||||
A) |
|
- |
35±22 (265) |
98 ± 26 (271) |
8 ± 5 (297) |
13 ± 5 (297) |
33 ± 20 (282) |
|
|
± |
20 ± 19 (239) |
90 ± 25 (252) |
11 ± 4 (283) |
34 ± 15 (280) |
27 ± 13 (308) |
B) This study |
|
- |
42 ± 14 (9) |
126 ± 24 (12) |
11 ± 5 (9) |
13 ± 2 (9) |
24 ± 8 (9) |
|
|
± |
20 ± 19 (8) |
104 ± 27 (11) |
12 ± 4 (9) |
24 ± 6 (12) |
28 ± 10 (12) |
Positive controls |
|||||||
EMS |
2µL |
- |
440 |
- |
- |
- |
- |
MMS |
1µL |
- |
- |
4860 |
- |
- |
- |
9 AA |
100 |
- |
- |
- |
103 |
- |
- |
Anthragallol |
50 |
- |
- |
- |
- |
47 |
81 |
2- Anthramine |
1 |
± |
824 |
7600 |
488 |
6400 |
7600 |
D and C Red No. 27 |
10 |
- |
43 |
113 |
8 |
12 |
14h |
|
± |
7 |
114 |
16h |
30 |
24 |
|
50 |
- |
51 |
30 |
9 |
18 |
11 |
|
|
± |
6 |
98 |
12 |
19 |
23 |
|
100 |
- |
41 |
109 |
7 |
11 |
11h |
|
|
± |
16 |
90 |
5 |
19 |
24 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
The substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid Red 92) (CAS Number: 18472-87-2) is non mutagenic in the In vivo Mammalian Erythrocyte Micronucleus Test performed.
Link to relevant study records
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- secondary literature
- Justification for type of information:
- Data is from secondary source
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- Principles of method if other than guideline:
- In vivo Mammalian Erythrocyte Micronucleus Test of Acid Red 92 in NMRI mouse was conducted.
- GLP compliance:
- not specified
- Type of assay:
- micronucleus assay
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): Fuchsia Red WR 802177
- Molecular formula: C20H2Br4Cl4O5Na2
- Molecular weight: 829. 6398 g/mol
- Substance type: Organic
- Physical state: Red powder
- Purity: 99.5 area % (HPLC), 85 % (w/w) (NMR, calculated as sodium salt). - Species:
- mouse
- Strain:
- NMRI
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- No data available.
- Route of administration:
- intraperitoneal
- Vehicle:
- No data available.
- Details on exposure:
- No data available
- Duration of treatment / exposure:
- Exp.1-
24 hrs- 25, 50, 100 mg/kg
48 hrs-100 mg/kg
Exp.2-100 mg/kg
Exp.3- 200 mg/kg
Exp.4-150 mg/kg - Remarks:
- Doses / Concentrations:Exp.1- 24 hrs- 25, 50, 100 mg/kg 48 hrs-100 mg/kg Exp.3- 200 mg/kg Exp.4-150 mg/kg Basis:
- No. of animals per sex per dose:
- Total 24 animals-12 male and 12 female
Exp.1-25 mg/kg: 2male and 2 female
50 mg/kg: 2male and 2 female
100 mg/kg: 2male and 2 female
Exp.2-100 mg/kg: 2male and 2 female
Exp.3- 200 mg/kg: 2male and 2 female
Exp.4- 150 mg/kg: 2male and 2 female - Control animals:
- yes, concurrent vehicle
- Positive control(s):
- Positive controls: Cyclophosphamide
- Justification for choice of positive control(s): No data available
- Route of administration: No data available
- Doses / concentrations: No data available - Tissues and cell types examined:
- Micronucleates (MN) produced in the polychromatic erythrocytes were observed
- Details of tissue and slide preparation:
- No data available
- Evaluation criteria:
- Frequency of MN was noted
- Statistics:
- No data available
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- yes
- Vehicle controls validity:
- not specified
- Negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- RESULTS OF RANGE-FINDING STUDY
- Dose range: No data available
- Solubility: No data available
- Clinical signs of toxicity in test animals: No data available
- Evidence of cytotoxicity in tissue analyzed: No data available
- Rationale for exposure: No data available
- Harvest times: No data available
- High dose with and without activation: No data available
- Other: No data available
RESULTS OF DEFINITIVE STUDY
- Types of structural aberrations for significant dose levels (for Cytogenetic or SCE assay): No data available
- Induction of micronuclei (for Micronucleus assay): In the treated animals with 100 mg/kg a percentage of MN 0.150 (24h) and 0.110 (48h) was observed: these values, although higher than the control, had a p > 0.34. reduction of PCE was observed, thus indicating a cytotoxic effect of the test item in the bone marrow cells.
- Ratio of PCE/NCE (for Micronucleus assay): No data available
- Appropriateness of dose levels and route: No data available
- Statistical evaluation: No data available - Conclusions:
- The substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid Red 92) (CAS Number: 18472-87-2) is non mutagenic in the In vivo Mammalian Erythrocyte Micronucleus Test performed.
- Executive summary:
Genetic toxicity in vivo study was performed for the test chemical3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid Red 92) (CAS Number: 18472-87-2) on male and female NMRI mice. Acid red 92 was injected intraperitoneally in mice and the number of micronucleus formed in the polychromatic erythrocytes was noted.
The positive control (CPA) induced 2.030 % of MN in PCEs (0.08 % in the control animals: significance p 0.0040).
In the treated animals with 100 mg/kg a percentage of MN 0.150 (24h) and 0.110 (48h) was observed: these values, although higher than the control, had a p > 0.34. A reduction of PCE was observed, thus indicating a cytotoxic effect of the test item in the bone marrow cells.
Acid Red 92 is non mutagenicintheIn vivo Mammalian Erythrocyte Micronucleus Test performed.
In accordance with the CLP classification, 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid Red 92) (CAS Number: 18472-87-2)is non mutagenic.
Reference
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
The potential genotoxicity of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2) was evaluated using in vitro and in vivo assays. Various peer reviewed publications for target substance have been reviewed and are summarized to determine the mutagenic potential of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid:
In Vitro
The Ames Salmonella/mammalian-microsome assay was conducted by J.P. Brown et al. (Mutation Research, 66 ,181-185, 1979) to evaluate the bacterial mutagenicity of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) (CAS Number: 18472-87-2) .3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) did not showed any mutagenic activity at concentrations up to 100 μg/plate in S.typhimurium strains TA1535, TA100, TA1537, TA1538 and TA98 in the presence and absence of S9 metabolic activation system.
Further, gene toxicity study was published in a SCCNFP report (COLIPA n° C53, 2004). The study was performed using reverse mutation assay on Salmonella typhimurium strains TA 1535; TA 1537; TA 98; TA 100; TA 102 to evaluate the mutagenic potential of the test compound 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid at dosage levels of 1, 10, 100, 1000, 5000 μg/plate.
The result was found to be as follows:
TA 98: no increase in the no. mutants in all conditions;
TA 100:no increase in the no. mutants in all conditions; 5000 μg/plate toxic ± S 9;
TA 1535: a weak mutagenic activity in the presence of S9 of no relevance; toxic effect at the maximum dose with S9.
TA 1537: no increase in mutants in all conditions
TA 102: some increase with the dose in all conditions, but never reaching the double.
There is a very weak indication of induced mutagenicity. The study as such can be considered as negative. The test item is not mutagenic in this assay.
In addition, the in vitro Mammalian Cell Gene Mutation assay was performed to evaluate the mutagenic potential of the test compound 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid red 92) (CAS No. 18472-87-2) in the Mouse lymphoma L5178 cells (TK locus). The study was published in a SCCNFP report (COLIPA n° C53, 2004).
Two experiments were performed: at different concentration and for different time duration
Exp.1:-S9: 5, 10, 20, 30, 40 μg/ml (4 hours)
+S9: 5, 10, 20, 40, 60 μg/ml (4 hours)
Exp.2:-S9: 19.5, 39, 78, 156 μg/ml (24 hours).
In a preliminary experiment a relevant toxicity was observed at 39.1 μg/ml in all conditions at 4 h of treatment; in the 24 h of treatment a severe toxicity was observed at 312.5 μg/ml. The doses for the main experiments were chosen on the base of a preliminary toxicity experiment. In treated cell a mutagenic effect observed in one culture was not repeated in the replicate culture. Therefore under the test conditions tested the test item 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid red 92) (CAS No. 18472-87-2) was considered to be non mutagenic in this assay.
Also, the gene toxicity study was conducted by K. LAURIE MAUS et al. (Mutation Research, 91 (1981) 315 -320) for the test compound 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Phloxine B) in Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538 by the Salmonella/microsome test with preincubation. The assay was carried out at 1000 µg/µmoles in duplicate and the number of His+revertants was calculated. Phloxine B was found to be negative (with and without S9 metabolic activation system) in this assay.
Moreover, K. LAURIE MAUS et al. (Mutation Research, 91 (1981) 315 -320) conducted gene toxicity study for the test compound Phloxine B in Escherichia coli strain WP2 by E. coli suspension test and by microtitre fluctuation assay.
The E. coli suspension assay was carried out at a concentration of 15,30 and 45 µg/ml in duplicate and the number of prototrophs /plate was calculated. Phloxine B was found to be negative in Escherichia coli strain WP2 by suspension test.
Whereas Escherichia coli microtitre fluctuation assay was carried out at a concentration range in duplicate and positive wells were scored on the basis of turbidity. The test compound Phloxine B is negative in Escherichia coli strain WP2 by microtitre fluctuation assay. Hence, it can be concluded that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9 -yl)benzoic acid (Phloxine B) (CAS No. 18472-87-2) is non genotoxic.
Based on the data summarized above, the test material does not classify as a gene mutant in vitro.
In Vivo
The genetic toxicity in vivo study was performed for the test chemical3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid Red 92) (CAS Number: 18472-87-2) on male and female NMRI mice.(published in a SCCNFP report (2004)). Acid red 92 was injected intraperitoneally in mice and the number of micronucleus formed in the polychromatic erythrocytes was noted.
The positive control (CPA) induced 2.030 % of MN in PCEs (0.08 % in the control animals: significance p 0.0040).
In the treated animals with 100 mg/kg a percentage of MN 0.150 (24h) and 0.110 (48h) was observed: these values, although higher than the control, had a p > 0.34. A reduction of PCE was observed, thus indicating a cytotoxic effect of the test item in the bone marrow cells.
3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (Acid Red 92) is non mutagenic in the In vivo Mammalian Erythrocyte Micronucleus Test performed.
Based on the above mentioned in vitro and in vivo studies for target substance, by applying weight of evidence approach and according to CLP classification criteria, it can be concluded that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2) is non genotoxic.
Justification for classification or non-classification
The results of several mutagenicity studies in vitro and in vivo shows that no classification for mutagenicity according to CLP regulation is warranted for 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2).
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