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EC number: 248-460-3 | CAS number: 27441-70-9
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
The test item resulted to be non-mutagenic in the AMES test performed.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- From November 12, 1985 to February 06, 1986
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- test procedure in accordance with generally accepted scientific standards and described in sufficient detail
- Justification for type of information:
- The read across approach is detailed into the document attached to the IUCLID section 13.
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium, other: TA 100, TA 1535, TA 1537, TA 1538, TA 98
- Details on mammalian cell type (if applicable):
- Bacteria are grown overnight in nutrient broth (25 g Oxoid Nutrient Broth No 2/liter) at 37 °C. The suitable amount of bacteria in the cell suspension is thecked by nephelometry. For inoculation, stock cultures which are stored at -80 °C, are used. The comound is tested with the strains Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538 and E. coli WP2uvrA. Identification of the different bacterial strains is performed periodically.
- Species / strain / cell type:
- E. coli WP2 uvr A
- Details on mammalian cell type (if applicable):
- Bacteria are grown overnight in nutrient broth (25 g Oxoid Nutrient Broth No 2/liter) at 37 °C. The suitable amount of bacteria in the cell suspension is thecked by nephelometry. For inoculation, stock cultures which are stored at -80 °C, are used. The comound is tested with the strains Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538 and E. coli WP2uvrA. Identification of the different bacterial strains is performed periodically.
- Metabolic activation:
- with and without
- Metabolic activation system:
- rat liver homogenate
- Test concentrations with justification for top dose:
- EXPERIMENT 1: 0, 0.8, 4, 20, 100 and 500 µg/plate
EXPERIMENT 2: 0, 4, 20, 60, 100, 200 and 500 µg/plate
EXPERIMENT 3: 0, 0.8, 4, 20, 100 and 500 µg/plate - Vehicle / solvent:
- - Solvent(s) used: DMSO
- Untreated negative controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- 2-nitrofluorene
- sodium azide
- benzo(a)pyrene
- other: M-methyl-N'-nitro-N-nitrosoguanidine // 2-Aminoanthracene
- Details on test system and experimental conditions:
- Top agar is prepared for the Salmonella strains by mixing 100 ml agar (0.6 % agar, 0.5 % NaCl) with 10 ml of a 0.5 mM histidine-biotin solution. With E. coli histidine is replaced by trypbophan (2.5 ml, 0.5 mM). The following ingredients are added (in order) to 2 ml of motten top agar at 45 °C: 0.1 ml of an overnight nutrient broth culture of the bacterial tester strain, 0.1 ml test compound solution, 0.5 ml S9 Mix (if required) or buffer
After mixing, the liquid is poured into a petridish with minimal-agar (1.5 % agar, Vogel-Bonner E medium with 2 % glucose). After incubation for 48 to 72 hours at 37 °C in the dark, colonies (his+ revertants) are counted.
CITOTOXICITY - DOSE RANGE FINDING
The first test was performed with all tester strains using three plates per dose to get information on mutagenicity and toxicity for calculation of an appropriate dose range. A reduced rate of spontaneously occuring colonies as well as visible thinning of the bacterial lawn were used as indicator for toxicity. Thinning of the bacterial lawn was controlled microscopically.
In combination with the first and third experiment, toxicity testing was performed as follows: 0.1 ml of the different dilutions of the test compound were thoroughly mixed with 0.1 ml of 10^-6 dilution of the overnight culture of TA 100 and plate with histidine and biotin rich top agar (3 plates per dose). In addition, toxicity testing was also performed in TA 98 and TA 1538 in the second experiment. The solvent control is compared with the number of colonies per ptate in the presence of the test compound.
PREPARATION OF LIVER HOMOGENATE FRACTION (S9)
Male Sprague Dawley rats (200 - 300 g) receive a single intraperitoneal injection of Aroclor t254 (500 mg/kg bodyweight) 5 days before sacrifice. Preparation is performed at 0 to 4 °C using-cold steiile solution and glassware. The livers from at least 5 - 6 animals are removed and pooled, washed in 150 mM KCI (ca 1 ml/g wet livers).
Sufficient S9 fraction is thawed immediately before each test at room temperature. One volume of S9 fraction is mixed with 9 volumes of the S9 cofactor solution and kept on ice until used. The concentrations of the different compounds in the S9 Mix are: 8 mM MgCl2, 33 mM KCl, 5 mM glucose-6-phosphate, 4mM NADP+, 100 mM phosphate buffer pH 7.4. - Species / strain:
- S. typhimurium, other: TA 100, TA 1535, TA 1537, TA 1538, TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- The test compound did not cause a significant increase in the number of revertant colonies with any of the tester strains neither in the absence nor presence of S9 Mix in two independent experiments. A very small increase in the number of revertant colonies just below the level of toxicity was obtained with TA 1538 and TA 98 in the presence of exogenous metabolic activation. Therefore the test was repeated with TA 1538 and TA 98 in the presence of S9 Mix.
The small increases in the number of revertant colonies never exeeded 2 times the control values and were of no toxicological significance.
TOXICITY TEST
The test compound was tested at doses of 4 to 10000 µg/plate and proved to be toxic to the bacteria at doses of 20 or 100 µg/plate. Thinning of the bacterial lawn and a reduction in the number of colonies has been observed at these doses. Therefore the test could be peformed only with reduced sensitivity. Visible precipitation of the test compound on the plates has been observed at 20 µg/plate.
For mutagenicity testing 500 µg/plate was chosen as the highest dose.
STERILE CHECKS AND CONTROL PLATES
Sterility of S9 Mix and the test compound was indicated by the absence of contamination on the test material and S9 Mix sterility check plates. Control plates (background control and positive controls) gave the expected number of colonies. - Conclusions:
- The test item resulted to be non-mutagenic in the AMES test performed.
- Executive summary:
The substance was tested for mutagenicity with the strains TA 100, TA 1535, 1538, TA 98 of Salmonella typhimurium and Escherichia coli WP2uvrA.
The rnutagenicity studies were conducted in the absence and in presence of a metabolizing system derived from rat liver homogenate. A dose range of 5 different doses from 0.8 µg/plate to 500 µg/plate was used. Control plates without mutagen showed that the number of colonies was similar to that described in the literature.
All the positive control compounds gave the expected increase in the number of revertant colonies.
The test compound proved to be toxic to most of the bacterial strains at 20 µg/ptate. On the basis of the preliminary test results the top dose level did not exceed 500 µg/plate.
In the absence of the metabolic activation system the test compound did not show a dose dependent increase in the number of revertants in any of the bacterial strains. Also in the presence of a metabolic activation system, treatment of the cells with test item did not result in relevant increases in the number of revertant colonies. Very slight increased nunbers of revertants with the strain TA 1538 and TA 98 were of no toxicological significance.
Summarizing, it can be stated that the test compound is not mutagenic in the bacterial test systems neither with nor without exogenous metabolic activation at the dose levels investigated.
Conclusion
The test item resulted to be non-mutagenic in the AMES test performed.
Reference
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
There are no specific information about the gene mutation in bacteria of Fluorescent Brightener 312, thus the available information on the structural analogous Similar Substance 02 have been taken into consideration; the read across approach can be considered as appropriate and suitable to assess the property under investigation (details about the approach are reported into the IUCLID section 13).
IN VITRO GENE MUTATION ASSAY IN BACTERIA
The substance was tested for mutagenicity with the strains TA 100, TA 1535, 1538, TA 98 of Salmonella typhimurium and Escherichia coli WP2uvrA.
The rnutagenicity studies were conducted in the absence and in presence of a metabolizing system derived from rat liver homogenate. A dose range of 5 different doses from 0.8 µg/plate to 500 µg/plate was used. Control plates without mutagen showed that the number of colonies was similar to that described in the literature.
All the positive control compounds gave the expected increase in the number of revertant colonies.
The test compound proved to be toxic to most of the bacterial strains at 20 µg/ptate. On the basis of the preliminary test results the top dose level did not exceed 500 µg/plate.
In the absence of the metabolic activation system the test compound did not show a dose dependent increase in the number of revertants in any of the bacterial strains. Also in the presence of a metabolic activation system, treatment of the cells with test item did not result in relevant increases in the number of revertant colonies. Very slight increased nunbers of revertants with the strain TA 1538 and TA 98 were of no toxicological significance.
It was concluded that the test compound is not mutagenic in the bacterial test systems neither with nor without exogenous metabolic activation at the dose levels investigated.
Justification for classification or non-classification
According to the CLP Regulation (EC ) No 1272/2008, for the purpose of the classification for germ cell mutagenicity, substances are allocated in one of two categories in consideration of the fact that they are:
- substances known to induce heritable mutations or to be regarded as if they induce heritable mutations in the germ cells of humans or substances known to induce heritable mutations in the germ cells of humans or
- substances which cause concern for humans owing to the possibility that they may induce heritable mutations in the germ cells of humans.
The available information suggest that test substance did not show any reasons of concern from the genotoxicity point of view.
In conclusion, the substance does not meet the criteria to be classified for genetic toxicity according to the CLP Regulation (EC) No 1272/2008.
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