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EC number: 208-719-3 | CAS number: 539-48-0
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Ames:
One study in 1998 shows negative to S. typhimurium (TA98, 100, 1535, 1537) and E. coli (WP2) strains.
Another study in 1997 shows positive to S. typhimurium TA104, and negative to other S. typhimurium (TA98, 100, 102, 1535, 1537) and E. coli (WP2uvrA, pKM101) strains.
In vitro chromosome aberration:
One study in 2000 shows negative result to CHL cells.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- From 1998-04-14 to 1998-04-27
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- GLP compliance:
- no
- Type of assay:
- bacterial reverse mutation assay
- Specific details on test material used for the study:
- Purity: 99.67%
- Target gene:
- Salmonella typhimurium: histidine
Escherichia coli: tryptophan - Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- Dose-finding study: 10, 50, 100, 500, 1000 and 5000 μg/plate
Main study: 78.1, 156, 313, 625, 1250, 2500 and 5000 μg/plate - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- no
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- For all strains with S9 mix
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- no
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 2-nitrofluorene
- Remarks:
- For strain TA98 without S9 mix
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- no
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide
- Remarks:
- For strain TA100 and WP2 without S9 mix
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- no
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- For strain TA1535 without S9 mix
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- no
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- For strain TA1537 without S9 mix
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
- Cell density at seeding (if applicable): 1E+09 cells/mL
DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 hours
NUMBER OF REPLICATIONS: 2 - Evaluation criteria:
- The test result are reported positive if the (mean) revertant colony count on the plate treated with the test substance is twice or more the (mean) spontaneous revertant colony count determined in the negative control group and apparent dose-dependence is found between the dose of test substance and the revertant colony count. The test result is reported negative in all other cases.
- Key result
- Species / strain:
- bacteria, other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Results obtained in the present study indicated that the test substance shows no mutagenic potential since the revertant colony counts determined for each tester strain in both the absence and presence of S9 mix were less than twice the spontaneous revertant colony count determined in the negative control group.
The test substance showed antibacterial activity against TA100, TA98, TA1535 and TA1537 at 2500 μg/plate and higher concentrations in both the presence and absence of S9 mix and against WP2 at 5000 μg/plate in the absence of S9 mix and 1250 μg/plate and higher concentrations in the presence of S9 mix. - Conclusions:
- The test substance shows no mutagenic potential to S. typhimurium and E. coli strains.
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- test procedure in accordance with national standard methods
- Qualifier:
- according to guideline
- Guideline:
- other: Guideline of the Ministry of Health, Labour and Welfare, Japan
- Deviations:
- no
- GLP compliance:
- no
- Type of assay:
- bacterial reverse mutation assay
- Specific details on test material used for the study:
- Lot No.: FBX01
Purity: 99.5 % - Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Details on mammalian cell type (if applicable):
- CELLS USED
- Source of cells: Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 were obtained from Dr. B.N. Ames (University of California), and Escherichia coli strains WP2/pKM101 and WP2uvrA/pKM101 were from Dr. M. Ishizawa (Kyushu University)
- Culturation: inoculated to nutrient broth and incubated for 10 h1's at 37°C in a shaking water bath befo1'e starting the test.
MEDIA USED
- Type and identity of media including CO2 concentration if applicable:
- Properly maintained: [yes/no]
- Periodically checked for Mycoplasma contamination: [yes/no]
- Periodically checked for karyotype stability: [yes/no)
- Periodically 'cleansed' against high spontaneous background: [yes/no] - Species / strain / cell type:
- other: S. typhimurium TA 102, TA 104 and E. coli WP2uvr
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- Experiment 1: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2uvrA: 0.0763, 0.305, 1.22, 4.88, 19.5, 78.1, 313, 1250, 5000 µg/plate
Experiment 2: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2uvrA: 19.5, 39.1, 78.1, 156, 313, 625, 1250, 2500 µg/plate
Experiment 3: S. typhimurium TA 102, TA 104 and E. coli WP2uvrA/pKM101: 0.0763, 0.305, 1.22, 4.88, 19.5, 78.1, 313, 1250, 5000 µg/plate
Experiment 4: S. typhimurium TA 102, TA 104 and E. coli WP2uvrA/pKM101: 19.5, 39.1, 78.1, 156, 313, 625, 1250, 2500 µg/plate - Vehicle / solvent:
- Vehicle used: Distilled H2O
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-(2-furyl)-3-(5-nitro-2-furyl)acrγlamide (AF2)
- Remarks:
- TA100, TA98 and WP2urA/pKM101 without S9 mix
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- TA1535 without S9 mix
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- TA1537 without S9 mix
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- WP2/pKM101 without S9 mix
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- TA98, TA100, TA102, TA104, TA1535, TA1537, WP2/pKM101 and WP2uvrA/pKM101 with S9 mix
- Details on test system and experimental conditions:
- - Preincubation Method:
The test compound dissolved in 0.05 or 0.1 mL of solvent was supplemented with 0.5 mL of S9 mix (metabolic activation method) or 0.1 M phosphate buffer pH 7.4 (direct method) and 0.1 mL of tester strains which had been cultured in nutrient broth. The mixture was incubated for 20 min at 37°C, and then mixed with 2 mL of molten top algar containing 0.05μmol/ mL of L-histidine and biotin for the Salmonella test (for the E. coli test 0.05μmol / mL of- L-tryptophan was used instead of L-histidine and biotin). Then the top agar mixture was immediately poured onto a 30 mL of Vogel-Bonner minimal glucose agar medium plate. A plate was kept on the flat table and top agar mixture was solidified. All plates were incubated for 48 hrs at 37 °C and the numbers of revertant colonies were scored. Tests on all test chemicals were done in duplicate plates, and those on positive control and solvent control were done in duplicate and quadruplicate plates, respectively.
- Determination of Toxicity (Growth Inhibition):
The growth of the background lawn of tester strains on each plate was examined under a stereo microscope. - Evaluation criteria:
- The chemicals were considered to be mutagenic when a dose-related increase in revertant colony count was observed and the number of revertant colonies per plate with the test substance was more than twice that of the negative control and when a reproducibility of test result was observed.
- Key result
- Species / strain:
- S. typhimurium, other: TA104
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- other: S. typhimurium TA98, TA100, TA102, TA1535, TA1537, E. coli WP2uvrA and WP2uvrA/pKM101
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Five experiments were conducted, the test item showed negative results to S. typhimurium TA98, TA100, TA102, TA1535, TA1537, E. coli WP2uvrA and WP2uvrA/pKM101 strains with and without metabolic activation, cytotoxicity observed at 2500 and 5000 μg/plate levels.
For TA 104, three experiments (3, 4, 5) were conducted with and without S9 mix. It showed positive result in experiment 3 with S9 only at 1250 μg/plate, and in experiment 5 with/without S9 at 800 and 1000 μg/plate levels, but negative at levels above 1000 μg/plate. Cytotoxicity observed at 5000 μg/plate level. The increase of revertant colony count is not dose-related. - Conclusions:
- The test item showed positive to S. typhimurium TA104, and negative to other S. typhimurium (TA98, 100, 102, 1535, 1537) and E. coli (WP2uvrA, pKM101) strains.
- Executive summary:
Bacterial mutagenicity test of test item was performed according to the guideline of the Ministry of Health, Labour and Welfare, Japan.
S. typhimurium TA98, TA100, TA102, TA104, TA1535, TA1537, E. coli WP2uvrA and WP2uvrA/pKM101 were tested with or without S9 mix.
Five experiments were conducted, the test item showed negative results to S. typhimurium TA98, TA100, TA102, TA1535, TA1537, E. coli WP2uvrA and WP2uvrA/pKM101 strains, cytotoxicity observed at 2500 and 5000 μg/plate levels.
For TA 104, three experiments (3, 4, 5) were conducted with and without S9 mix. It showed positive result in experiment 3 with S9 only at 1250 μg/plate, and in experiment 5 with/without S9 at 800 and 1000 μg/plate levels, but negative at levels above 1000 μg/plate. Cytotoxicity observed at 5000 μg/plate level. The increase of revertant colony count is not dose-related.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- test procedure in accordance with national standard methods
- Qualifier:
- according to guideline
- Guideline:
- other: "Standard for Chromosomal Aberration Test in Cultured Mammalian Cells" of the Notification No.652 of the Labour Standards Bureau, Ministry of Labour, Japan
- Version / remarks:
- September 29, 1997
- Deviations:
- no
- GLP compliance:
- no
- Type of assay:
- in vitro mammalian chromosome aberration test
- Specific details on test material used for the study:
- Lot No.: FBX01
Purity: 99.5% - Species / strain / cell type:
- other: Chinese hamster lung cells
- Details on mammalian cell type (if applicable):
- CELLS USED
- Source of cells: supplied by Dr. M. Ishidate, Jr. in 1985
- Modal number of chromosomes: 25
- Doubling time: approximately 15 hours
MEDIA USED
- Type and identity of media including CO2 concentration if applicable: Eagle's MEM supplemented with 10% heat-inactivated (56°C, 30 min) calf serum - Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- 24 and 48 h treatment: 0.015, 0.03, 0.06, 0.09, 0.12, 0.15 mg/mL
6h treatment: 0.05, 0.1, 0.2, 0.3, 0.4 mg/mL - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: the test substance was soluble in DMSO and insoluble in water. - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- Remarks:
- with and without S9
- Details on test system and experimental conditions:
- - Short-period treatment with and without Metabolic Activation (6 hours treatment):
Twenty thousands cells were seeded in culture vessel (with 5 mL of culture medium) and cultured for three days, then culture medium was changed. Cells were simultaneously treated with and without S9 mix and the test substance solution for 6 hours. Then, the test substance mixture was changed to fresh culture medium. After 18 hours further culture, rates of cell growth inhibition were determined.
- Continuous Treatment without Metabolic Activation (24 and 48 hours treatment):
Twenty thousands cells were seeded in a culture vessel (with 5 mL of culture medium), and cultured for three days. Then culture medium was changed and the test substance solution was added to a culture medium. After 24 or 48 hours treatment, rates of cell growth inhibition were determined.
- Determination of Cell Growth Inhibition Rate
The medium was discarded and the cells were washed with physiological saline. Ethanol was added to fix the cells which were then stained with 0.1% crystal violet. After washing and drying, each dish was placed under a cell densitometer to measure the color absorption value (550nm). The cell growth index was calculated with the negative control being 100% and Petri dish without cells being 0%.
- Slide preparation:
Two hours before the end of culture, the cells were treated with 0.2μg/mL Colcemid. After finishing culture, they were dissociated with trypsin (0.1% trypsin + 0.036% EDTA) and centrifuged (1000 rpm, 5 min) to collect cells. After removal of the supernatant, the cells were incubated in a 75 mM hypotonic KCl solution for 20 minutes at 37 °C. The cells were then fixed three times with acetic acid-ethanol (1:3) and spread onto clean glass slides. Each slide was stained with Giemsa solution (2.5% at pH 6.8) for 12 minutes after air-drying.
- Observation:
The number of cells with structural aberrations in 100 well-spread metaphase cells were counted per each culture vessel. The incidence of polyploid cells were recorded simultaneously. - Evaluation criteria:
- Negative: frequencies of structural aberrations and of polyploidy less than 5%
Equivocal: frequencies of structural aberrations and of polyploidy from 5% to less than 10%
Positive: frequencies of structural aberrations and of polyploidy 10% or more - Key result
- Species / strain:
- other: Chinese hamster lung cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Negative result got for test item in chromosomal aberration test in CHL.
- Executive summary:
In vitro mammalian chromosome aberration test for test item was performed using lung cells of a newborn Chinese hamster according to "Standard for Chromosomal Aberration Test in Cultured Mammalian Cells" of the Notification No.652 of the Labour Standards Bureau, Ministry of Labour, Japan. The dose level were 0.015, 0.03, 0.06, 0.09, 0.12, 0.15 mg/mL for 24 and 48 h treatment without S9 mix activation, 0.05, 0.1, 0.2, 0.3, 0.4 mg/mL for 6h treatment with and without S9 mix activation. The recovery time forshort-period treatment (6h) was 18 h.
Negative result was given under the test condition.
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Two Ames studies are available.
One showed negative to S. typhimurium (TA98, 100, 1535, 1537) and E. coli (WP2) strains both with and without metabolic activation.
In another study, five experiments were conducted, the test item showed negative results to S. typhimurium TA98, TA100, TA102, TA1535, TA1537, E. coli WP2uvrA and WP2uvrA/pKM101 strains with and without metabolic activation, cytotoxicity observed at 2500 and 5000 μg/plate levels. For TA 104, three experiments (3, 4, 5) were conducted with and without S9 mix. It showed positive result in experiment 3 with S9 only at 1250 μg/plate, and in experiment 5 with/without S9 at 800 and 1000 μg/plate levels but negative at levels above 1000 μg/plate. Cytotoxicity observed at 5000 μg/plate level. The increase of revertant colony count is not dose-related.
One in vitro chromosome aberration study showed negative result to CHL cells.
All of these in vitro studies were non-GLP and according to national standard methods which similar to OECD Guidelines.
The results of two Ames studies are consistent as both of them showed negative result to same strains, except for the positive result on strain TA104 in the second study. The positive result on strain TA104 were observed in relatively medium levels but not at high levels, and the increase of revertant colony count is not dose-related, also considering the evidence of cytotoxicity at high doses, we cannot conclude a positive result simply.
Justification for classification or non-classification
In accordance with Regulation (EC) No. 1272/2008 Table 3.5.1, and Guidance on application of CLP criteria section 3.5.2.4, classification Category 2 is based on positive evidence from in vivo/vitro mammalian experiments, in vitro results can only lead to a Category 2 mutagen classification in a case where there is support by chemical structure activity relationship to known germ cell mutagens.
This substance showed positive in Ames study but negative in in vitro mammalian chromosome aberration study, and there is no evidence of analogue chemical which known as mutagen, so this substance shall not be classified for the mutagenicity endpoint.
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