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EC number: 279-317-3 | CAS number: 79828-43-6
- Life Cycle description
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- Endpoint summary
- Appearance / physical state / colour
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- Endpoint summary
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- 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
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- Additional ecotoxological information
- Toxicological Summary
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- Acute Toxicity
- Irritation / corrosion
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- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
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- Additional toxicological data
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
In a bacterial reverse mutation assay conducted according to OECD Guideline 471 and 472 in compliance with GLP, Substance S70767 (FAT 40821) gave a positive response (mutagenic) in S.typhimurium strains TA1535, TA1537, TA98 and TA100 and in E.coli strain WP2P, in both the presence and absence of S9. Limited activity was observed in E.coli strain WP2PuvrA in both the presence and absence of S9.
A similar study performed to determine the mutagenicity of FAT 40210 in bacterial cells according to OECD Guideline 471 (Bacterial Reverse Mutation Assay) showedFAT 40210/A was mutagenic for S.typhimurium strains TA 1537, TA 98 and TA 100. No mutagenic effect was observed with the strain TA 1535.
The source chemical, FAT 40821 was found to be not mutagenic in a study conducted according to OECD Guideline 476 (In vitro Mammalian Cell Gene Mutation Test).No substantial and reproducible dose dependent increase in mutant colony numbers was observed in the main experiments of this study.
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:
- 1994
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
- Principles of method if other than guideline:
- None
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- Salmonella typhimurium histidine (his) reversion system
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- other: his(+), uvrB(-), rfa(+), R factor(+)for TA 98 and TA 100; R factor(-)for TA 1535 and TA 1537
- Species / strain / cell type:
- E. coli, other: E.coli WP2P and WP2PuvrA
- Additional strain / cell type characteristics:
- other: uvrA mutation
- Metabolic activation:
- with and without
- Metabolic activation system:
- S-9 mix (Rat liver enzymes)
- Test concentrations with justification for top dose:
- 100 - 5000 µg/plate
- Vehicle / solvent:
- water
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-Aminoanthracene
- Remarks:
- for all strains with S9
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- TA 100, TA 1535 without S9
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: Acridine Mutagen ICR191
- Remarks:
- TA 1537 without S9
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: Daunomycin HCl
- Remarks:
- TA 98 without S9
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- E. coli WP2P without S9
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- N-ethyl-N-nitro-N-nitrosoguanidine
- Remarks:
- E. coli WP2P uvrA· without S9
- Details on test system and experimental conditions:
- A solution of the test compound (as supplied) was prepared in sterile (filtered: Milli-Q/RO) water to yield a w/v concentration of 50.00 mg/ml.
This was then diluted with water to give additional solutions of 25 mg/ml, 10 mg/ml, 5.0 mg/ml, 2.0 mg/ml and 1.0 mg/ml. Fresh stock solutions and dilutions were prepared as necessary for each experiment.
Water was also included as the negative (solvent) control.
The mutagenicity assays were conducted using the Salmonella bacterial mutation assay as described by Maron and Ames (1983), as updated by the United Kingdom Environmental Mutagen Society's sub-committee on Guidelines for Mutagenicity Testing (Gatehouse et al, 1990) (see Appendix A). The four Salmonella tester strains (TA1535, TA1537, TA98 and TA100) and two E.coli strains (WP2P and WP2PuvrA) used in this assay have been fully described in the literature (Ames et al 1975, and references therein; Venitt and Crofton-Sleigh, 1979).
The sample of Substance S70767 was assayed twice using the standard plate incorporation protocol over a dose range of 5000 - 100 µg per plate using all six strains (in a total of three separate experimental runs), both in the presence and absence of a liver S9-mix prepared from Phenobarbital/ beta-Naphthaflavone-induced Alderley Park (Alpk:APfSD) rats.
The incubation period for each experiment was 3 days (at 37°C).
For each experiment, positive control compounds were tested to validate the bacterial strains and to confirm the activity of each batch of S9-mix used.
Revertant colonies were counted using an automated electronic colony counter (AMS 40-10 Image Analyser fitted with appropriate software, Analytical Measuring Systems Ltd). - Evaluation criteria:
- Test data from individual experiments are considered valid if:
a) the concurrent solvent control data are acceptable;
b) the positive control data show unequivocal positive responses;
c) at least the lowest test compound dose shows no evidence of toxicity, and at least three test doses show no significant toxicity (ie significant loss of background growth and/or reductions in colony numbers).
Failure of one or more tester strain/59 combinations does not invalidate the data for the remainder of a concurrent experiment.
A positive response in a (valid) individual experiment is achieved when one or both of the following criteria are met:
a) a statistically significant dose-related increase in the mean number of revertant colonies is obtained;
b) a two-fold or greater increase in the mean number of revertant colonies (over that observed for the concurrent solvent control plates) which is
statistically significant, is observed at at least one dose level.
A negative result in a (valid) individual experiment is achieved when:
a) There is no statistically significant dose-related increase in the mean number of revertant colonies per plate observed for the test compound; and
b) in the absence of any such dose response, no increase in colony numbers is observed (at any test dose) which exceeds 2x the concurrent solvent control.
For a positive response in an individual experiment to be considered indicative of an unequivocal positive, ie mutagenic, result for that strain/59 combination, then the observed effect(s) must be consistently reproducible. - Statistics:
- All derived calculations (ie mean colony count/plate; standard deviation, etc) shown in the results tables were carried out by computer.
Counts from contaminated plates are not included in these calculations.
An assessment of statistical significance was carried out using a one-tailed Student's t-test (Ehrenberg 1984). The corresponding probability for each dose level was derived by computer using the appropriate degrees of freedom. Values of p<0.01 are treated as significant, with values of 0.01<=p<0.05 being indicative of a possible effect. - Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- not specified
- Remarks:
- but tested up to limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- not specified
- Remarks:
- but tested up to limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- ambiguous
- Cytotoxicity / choice of top concentrations:
- not specified
- Remarks:
- but tested up to limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- In two separate assays, the compound induced significant, reproducible increases in the observed number of revertant colonies in strains TA1535, TA1537, TA98, TA100 and WP2P both in the presence or absence of an auxiliary metabolising system (S9). Limited activity was observed in strain WP2PuvrA both in the presence and absence of S9.
The positive controls for each experiment induced the expected responses, indicating the strains were responding satisfactorily in each case. - Conclusions:
- The test substance gave a positive, ie mutagenic response in S.typhimurium strains TA1535, TA1537, TA98 and TA100 and in E.coli strain WP2P in both the presence and absence of S9. Limited activity was observed in E.coli strain WP2PuvrA in both the presence and absence of S9.
- Executive summary:
A study was performed to determine the mutagenecity of Substance S70767 according to OECD Guideline 471 (Bacterial Reverse Mutation Assay) and OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay). Substance S70767 has been evaluated in a bacterial mutagenicity assay (Gatehouse et al, 1990: based on Maron and Ames (1983)) using four strains of Salmonella typhimurium (TA1535, TA1537, TA98 and TA100) and two strains of Escherichia coli (WP2P and WP2PuvrA), following protocols complying with OECD Guideline Numbers 471 and 472 (OECD, 1983a and 1983b) and with the United Kingdom Department of Health Guidelines (DoH, 1989).
In two separate experiments, the compound induced significant, reproducible increases in the observed numbers of revertant colonies in all of the tester strains used, both in the presence or absence of an auxiliary metabolising system (S9). In each experiment, the positive controls responded as expected indicating that the assay was performing satisfactorily. Under the conditions of this assay, Substance S70767 therefore gave a positive, ie mutagenic response in S.typhimurium strains TA1535, TA1537, TA98 and TA100 and in E.coli strain WP2P in both the presence and absence of S9. Limited activity was observed in E.coli strain WP2PuvrA in both the presence and absence of S9.
- 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:
- 2005
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Principles of method if other than guideline:
- None
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- HPRT locus
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-mix
- Test concentrations with justification for top dose:
- 2.5, 5, 10, 20, 40 μg/ml Experiment I without S9-mix
39, 78, 156, 234, 312 μg/ml with S9-mix
19.5, 39, 78, 117, 156 μg/ml experiment II without S9-mix - Vehicle / solvent:
- deion.water
- Untreated negative controls:
- yes
- Remarks:
- culture medium
- Negative solvent / vehicle controls:
- yes
- Remarks:
- solvent
- True negative controls:
- yes
- Remarks:
- untreated cells
- Positive controls:
- yes
- Remarks:
- with and without S9
- Positive control substance:
- 7,12-dimethylbenzanthracene
- ethylmethanesulphonate
- Details on test system and experimental conditions:
- None
- Evaluation criteria:
- None
- Statistics:
- None
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 10 μg/ml and more without S9, 234 and above with S9
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks on result:
- other: strain/cell type: V79
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- FAT40821/A is considered to be non-mutagenic in this HPRT assay
- Executive summary:
In a GLP-compliant mammalian cell gene mutation test, performed according to OECD guideline 476, V79 cells of the Chinese hamster were exposed to the test substance FAT 40821 with and without metabolic activation to investigate the potential of the test substance to induce gene mutations at the HPRT locus. The assay was performed in two independent experiments. The cells were exposed to the test item for 4 hours in the first experiment with and without metabolic activation. The second experiment was solely performed in the absence of metabolic activation with a treatment period of 24 hours. No substantial and reproducible dose dependent increase in mutant colony numbers was observed in both main experiments. In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells. Therefore, FAT 40821 is considered to be non-mutagenic in this HPRT assay.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- an in vitro cytogenicity study in mammalian cells or in vitro micronucleus study does not need to be conducted because adequate data from an in vivo cytogenicity test are available
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (positive)
Genetic toxicity in vivo
Description of key information
FAT 40210 was tested for clastogenic potential in two micronucleus assays.
An GLP compliant in vivo study was performed to investigate the potential of FAT 40210/E to induce micronuclei in polychromatic erythrocytes (PCE) in the bone marrow of the mouse according to OECD guideline 474. The test article FAT 40210/E was investigated at 24 h preparation interval at 200, 670 and 2000 mg/kg b.w.(20 ml/kg b.w.); and 48 h preparation interval at 2000 mg/kg b.w.(20 ml/kg b.w.). FAT40210/E had no cytotoxic effects in the bone marrow and did not induced micronuclei by the micronucleus test with bone marrow cells of the mouse. FAT 40210 was considered to be non-mutagenic in the micronucleus assay according to CLP Regulation (Regulation EC No. 1272/2008).
In another in vivo study, test substance S70767 was tested in CD-1 mice at the maximum tolerated doses of 500 mg/kg in males and 320 mg/kg in females, based on the patterns of clinical observations and lethalities recorded over a four day observation period. Bone marrow samples were taken 24 and 48 hours after dosing. No statistically or biologically significant increases in the incidence of micronucleated polychromatic erythrocytes, over the vehicle control values, were seen in either sex at either of the sampling times investigated. Statistically significant reductions in the percentage of polychromatic erythrocytes, compared to the vehicle control values, were observed in females treated with Substance S70767 at the 24-hour sampling time and in males treated with Substance S70767 at the 48-hour sampling time, indicating that Substance S70767 or a metabolite may have had a cytotoxic effect on the bone marrow. The test system positive control, cyclophosphamide, induced statistically significant and biologically meaningful increases in micronucleated polychromatic erythrocytes, compared to the vehicle control values, thus demonstrating the sensitivity of the test system to a known clastogen. It is therefore concluded that Substance S70767, under the conditions of test, is not clastogenic in the mouse bone marrow micronucleus test.
Reactive Black 8 was found to be not clastogenic in both in vivo studies.
Link to relevant study records
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1996-1997
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- Deviations:
- no
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- other: EEC Directive 92/69, L383, Annex V, B12, dated December 29, 1992
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- micronucleus assay
- Species:
- mouse
- Strain:
- NMRI
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- Animals: Strain-NMRI; Source: BRL, CH-4414 Füllinsdorf;
Number of Animals: 36 males/36 females;
Initial Age at start of Acclimatization: 8-12 weeks;
Acclimatization: minimum 5 days;
Initial body weight at start of treatment: males mean value 31.0g (SD±2.5g), female mean value 23.4 g (SD±1.5g).
The animals were in healthy condition and under quarantine in the animal house of CCR for a minimum of five days after their arrival. During this period the animals did not show any signs of illness or altered behavior. The animals were distributed into the test groups at random and identified by cage number.
Environmental conditions
Husbandry: the animals were kept conventionally. The experiment was conducted under standard laboratory conditions;
Housing: single; Cage Type: Makrolon Type I, with wire mesh top (EHRET GmbH, D-79302 Emmendingen);
Bedding: granulated soft wood bedding (ALTRONIM, D-32791 Lage/Lippe);
Feed: pelleted standard diet, ad libitum ((ALTRONIM 1324, D-32791 Lage/Lippe);
Water: tap water, ad libitum, (Gemeindewerke, D-64380 Robdarf); E
nvironment: temperature 21±3 ℃, relative humidity 30-70 %, artificial light 6.00 a.m.-6.00 p.m. - Route of administration:
- oral: unspecified
- Vehicle:
- deionised water
- Details on exposure:
- test material: a single standard volume of 20ml/kg bogy weight orally (200, 670 and 2000 mg/kg b.w. respectively);
Vehicle control: orally, once, 20 ml/kg b.w.;
Positive control: orally, once, 40 mg/kg b.w.(20 mg/kg b.w.) - Duration of treatment / exposure:
- 24 h and 48 h
- Frequency of treatment:
- once
- Dose / conc.:
- 200 mg/kg bw/day (nominal)
- Dose / conc.:
- 670 mg/kg bw/day (nominal)
- Dose / conc.:
- 2 000 mg/kg bw/day (nominal)
- No. of animals per sex per dose:
- 5 per sex per dose
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- Cyclophosphamide
- Tissues and cell types examined:
- polychromatic erythrocytes
- Details of tissue and slide preparation:
- The animals were sacrificed by cervical dislocation. The femora were removed, the epiphyses were cut off and the marrow was flushed out with fetal calf serum, using a syringe. The cell suspension was centrifuged at 1500 rpm (390 X g) for 10 minutes and the supernatant was discarded. A small drop of the resuspended cell pellet was spread on a slide. The smear was air-dried and then stained with May-Grunwald (MERCK, D-64293 Darmstadt)/Giemsa (Gurr, BDH Limited Poole, Great Britain). Cover slips were mounted with EUKITT (KINDLER, D-79110 Freibur). At lease one slide was made from each bone marrow sample.
- Evaluation criteria:
- A test article is classified as mutagenic if it induces either a dose-related increase in the number of micronucleated polychromatic erythrocytes or a statistically significant positive response for at least one of the test points. A test article producing neither a dose-related increase in the number of micronucleated polychromatic erythrocytes nor a statistically significant positive response at any of the test points is considered non-mutagenic in this system. This can be confirmed by means of the nonparametric Mann-Whitney test. However, both biological and statistical significance should be considered together.
- Sex:
- male/female
- Genotoxicity:
- negative
- Remarks:
- did not induce micronuclei as determined by the micronucleus test in the bone marrow cells of the mouse
- Toxicity:
- no effects
- Remarks:
- None of the animals expressed toxic reactions treated with 2000 mg/kg b.w. and FAT 40210/E had no cytotoxic properties in the bone marrow.
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- valid
- Conclusions:
- FAT 40210/E was considered to be non-mutagenic in the micronucleus assay.
- Executive summary:
The study was performed to investigate the potential of FAT 40210/E to induce micronuclei in polychromatic erythrocytes (PCE) in the bone marrow of the mouse.
The following dose levels of the test article were investigated: 24 h preparation interval: 200, 670 and 2000 mg/kg b.w.(20ml/kg b.w.), 48 h preparation interval: 2000 mg/kg b.w.(20 ml/kg b.w.).
FAT40210/E had no cytotoxic effectiveness in the bone marrow and did not induced micronuclei by the micronucleus test with bone marrow cells of the mouse. FAT40210 was considered to be non-mutagenic in the micronucleus assay according to CLP Regulation (Regulation EC No. 1272/2008).
Reference
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Taking in vitro and in vivo study results into account, Reactive Black 8 was found to have mutagenic potential in bacterial cells. However, the read-across chemical (FAT 40821) was not mutagenic in a mammalian cell gene mutation assay in vitro (HPRT). Thus, Reactive Black 8, using the principles of read-across, can also be considered to be not mutagenic in mammalian cells in vitro. Further, Reactive Black 8 was not clastogenic in the micronucleus assays. Hence taking into consideration the available results, Reactive Black 8 can be regarded as non-mutagenic and non-clastogenic.
Justification for classification or non-classification
Based on the above stated assessment of the genotoxic potential, the test item (Ames test positive, in vitro gene mutation (similar substance) negative, two in vivo mammalian micronucleus tests in bone marrow cells negative) is deemed non-genotoxic and accordingly does not need to be classified according to CLP (REGULATION (EC) No 1272/2008 OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL) as implementation of UN-GHS in the EU.
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.
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