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EC number: 201-126-0 | CAS number: 78-59-1
- 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 majority of in vitro genotoxicity studies revealed clearly negative results.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- Restriction: only 4 Salmonella strains tested
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- 4 strains tested instead of 5
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254-induced rat or hamster liver S9
- Test concentrations with justification for top dose:
- TA 100 and TA 1535 : 0 - 10000 μg/plate
TA 98 and TA 1537: 0 - 3333 μg/plate - Vehicle / solvent:
- water
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- yes
- Remarks:
- potassium chloride
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- Strains TA 1535 and TA 100, without metabolic activation
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- yes
- Remarks:
- potassium chloride
- Positive controls:
- yes
- Positive control substance:
- other: 4-nitro-o-phenylenediamine
- Remarks:
- Strain TA 98, without metabolic activation
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- yes
- Remarks:
- potassium chloride
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- Strain TA 1537, without metabolic activation
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- yes
- Remarks:
- potassium chloride
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- All strains, with metabolic activation
- Details on test system and experimental conditions:
- - Number of replicates: 3 per dose level, repeated
- Solvent: H2O
- 3 investigations were performed:
1. without metabolic activation
2. with metabolic activation (Arochlor-1254 liver rats)
3. with metabolic activation (Arochlor-1254 liver hamster)
- Cells and test compound or solvent (water) were incubated for 20 minutes at 37 ºC in the presence of either S9 or buffer. After the addition of soft agar, the contents of each tube were poured onto minimal medium, and the plates were incubated at 37 °C for 48 hours.
- The analysis was performed twice, each in triplicates. - Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: -S9: TA100, 1535, 98 at concentrations >=1000 µg/plate; +S9 (rat and hamster): only TA1535 at 10000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- The test substance was not mutagenic in strains TA100, TA1535, TA1537, or TA98 of Salmonella Typhimurium in the presence or absence of Aroclor 1254-induced rat (r.) or hamster (h.) liver S9.
Differences were not significant.
Maximum number of revertants in the standard plate test:
TA100:
Vehicle Test substance (µg/plate)
-S9 +S9(r.) +S9(h.) -S9 +S9(r.) +S9(h.)
-----------------------------------------------------------
82±4.7 92±7.5 79±2.2 88±7.4(333) 90±2.2(100) 117±2.9(333)
TA1535:
Vehicle Test substance (µg/plate)
-S9 +S9(r.) +S9(h.) -S9 +S9(r.) +S9(h.)
---------------------------------------------------------
6±2.5 7±0.9 7±1.2 4±0.9(100) 9±2.8(333) 6±1.0(100)
TA1537:
Vehicle Test substance (µg/plate)
-S9 +S9(r.) +S9(h.) -S9 +S9(r.) +S9(h.)
----------------------------------------------------------
2±0.3 3±1.5 4±0.7 4±0.7(1000) 5±1.0(100) 6±2.0(1000)
TA98:
Vehicle Test substance (µg/plate)
-S9 +S9(r.) +S9(h.) -S9 +S9(r.) +S9(h.)
----------------------------------------------------------
10±2.0 11±2.9 17±1.0 12±2.3(100) 17±1.3(100) 15±0.9(333)
The analysis was performed twice, each in triplicate; because the results were similar, data from only one experiment are shown. - Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian cell gene mutation tests using the thymidine kinase gene
- Target gene:
- TK
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Metabolic activation system:
- aroclor 1242 and 1254 (2:1 mixture) induced rat liver microsomes
- Test concentrations with justification for top dose:
- Without activation
0.13; 0.18; 0.24; 0.32; 0.42, 0.56; 0.75; 1.0; 1.3 µL/mL
With activation
0.067; 0.089; 0.12; 0.16; 0.21; 0.28; 0.38; 0.50; 0.67; 0.89 µL/mL
Concentration values are calculated: original data are given in µL/mL; a density factor of 0.92 g/cm3 is assumed. - Vehicle / solvent:
- DMSO
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 7,12-dimethylbenzanthracene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4 h
- Expression time (cells in growth medium): 24 and 48 h
NUMBER OF REPLICATIONS: 2 - Evaluation criteria:
- positive: positive dose response and >= 1 of 3 highest doses with a mutant frequency 2-fold greater than background.
equivocal: no dose response but any dose with a mutant frequency 2-fold greater than background. - Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- True negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Qualifier:
- according to guideline
- Guideline:
- other: Clive and Spector, Mutat. Res. 44, 269-278 (1975) and Clive et al., Mutat. Res. 59, 61-108 (1979)
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian cell gene mutation tests using the thymidine kinase gene
- Target gene:
- TK
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- without
- Test concentrations with justification for top dose:
- 50 - 1600 µg/mL (details see Test Conditions)
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Details on test system and experimental conditions:
- Experiments were performed twice, and all doses were tested in duplicate, except the solvent control (DMSO), which was tested in quintuplicate. Cells (6 x 10E+5/mL) were treated for 4 hours at 37 ºC in medium, washed, resuspended in medium, and incubated for 48 hours at 37 ºC. After expression, 3 x 10E+6 cells were plated in medium supplemented with trifluorothymidine for selection of cells that were mutant at the thymidine kinase (TK) locus, and 600 cells were plated in nonselective medium to determine the percentage of viable cells.
Trial 1: 0, 50, 100, 200, 400, 800, 1600 µg/mL
Trial 2: 0, 400, 600, 800, 1000, 1200 µg/mL
Trial 3: 0, 200, 400, 600, 800, 1000 µg/mL - Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 1600 µg/mL
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- GENOTOXIC EFFECTS
Without metabolic activation: mutagenic. The test substance was toxic to the cultures only at moderately to high concentrations.
Significant increases in mutant fraction occurred in all three experiments, accompanied by a reduction of relative total growth (RTG).
The lowest effective concentration was 400 µg/mL in the first experiment, where there was apparently no reduction in RTG from the vehicle control level.
In the second experiment, only at 600 µg/mL there was evidence of toxicity.
The lowest effective concentration in this experiment was 800 µg/mL. However, the cloning efficiency was low in this experiment (results questionable). In the third experiment the lowest effective concentration was 800 µg/mL.
Tennant et al. (1997) judged the test substance as positive at concentrations >= 400 µg/mL without metabolic activation.
CYTOTOXICITY
Relative total growth reduced at 800, 1000 and 1200 µg/mL. - Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Qualifier:
- according to guideline
- Guideline:
- other: Galloway, S.M. et al., Environ Mol. Mutagen. 10 (10), 1-175, with a few modifications
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Metabolic activation system:
- aroclor 1254 induced rat liver
- Test concentrations with justification for top dose:
- without metabolic activation
50-1600 µg/mL
with metabolic activation
250-1500 µg/mL - Vehicle / solvent:
- serum-free culture medium
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- no
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- methylmethanesulfonate
- Details on test system and experimental conditions:
- Analysis: 100 or 200 cells were scored for each dose (cells with chromosome number lower than 19 or higher than 23 were excluded).
In the absence of S9, CHO cells were incubated with test compound or solvent for 8-10 hours at 37 ºC. Cells were then washed, and fresh medium containing colcemid (0.1 µg/mL) was added. After a further 2-3 hours of incubation, cells were harvested by mitotic shake-off, fixed, and stained in 6% Giemsa. In the presence of S9, cells were incubated with test compound or solvent for 2 hours at 37 ºC. Cells were then washed, medium was added, and incubation continued for 8-10 hours. Colcemid (0.1 µg/mL) was added for the last 2-3 hours of incubation; then cells were harvested and fixed as above.
2 independent experiments:
without S9 mix
50, 160, 500, 1600 µg/mL
250, 500, 1000, 1600 µg/mL
with S9 mix
250, 500, 1000 µg/mL
750, 1000, 1250, 1500 µg/mL - Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- ≥ 5000 µg/mL
- Vehicle controls validity:
- not applicable
- Untreated negative controls validity:
- not specified
- True negative controls validity:
- not applicable
- Positive controls validity:
- not specified
- Additional information on results:
- The test substance did not induce a significant increase of structural chromosomal aberrations in the presence or absence of S9 in Chinese hamster ovary cells.
Maximum number of cells with structural chromosomal aberrations (Abs) per 100 cells at given concentration of test substance:
Abs (µg/mL)
-S9 +S9
--------------------
2 (vehicle) 0 (vehicle)
5 (250) 2 (1500)
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
Negative results were seen in in vivo micronucleus tests and chromosomal abberration tests in mice.
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:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- GLP compliance:
- not specified
- Type of assay:
- mammalian erythrocyte micronucleus test
- Species:
- mouse
- Strain:
- CD-1
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Breeding Laboratory, Kingston, New York
- Age at study initiation: 6-8 weeks - Route of administration:
- intraperitoneal
- Vehicle:
- corn oil
- Details on exposure:
- Single intraperitoneal application of 496.8 mg/kg of the test substance as mixture with vehicle in 10 mL/kg bw.
- Duration of treatment / exposure:
- 12, 24, 48 hours
- Frequency of treatment:
- single
- Post exposure period:
- none
- Dose / conc.:
- 496.8 mg/kg bw/day (nominal)
- Remarks:
- = Maximum Tolerated Dose/LD20
- No. of animals per sex per dose:
- 5
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- triethylenemelamine
- Doses / concentrations: 0.25 mg/kg - Tissues and cell types examined:
- erythrocytes from femur bone marrow
- Details of tissue and slide preparation:
- Sampling times: 12, 24, 48 h post dosing.
Criteria for selection of Maximum Tolerated Dose (M.T.D.): LD20 (determined in a preliminary toxicity study) - Statistics:
- 1-way analysis of variance and Duncan's multiple range test (P <= 0.05)
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- not specified
- Remarks:
- Maximum Tolerated Dose (LD20) was applied
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- valid
Reference
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
In vitro studies
Several tests with S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 with and without S9 were negative (Mortelmans et al., 1986; Huntingdon, 1978; Huels AG, 1988).
The mouse lymphoma tests were primarily negative. In the presence of metabolic activation one positive result was obtained in one of the two testing facilities leading to an inconclusive overall result (Honma et al., 1999). Without metabolic activation some studies gave negative results (Microbiological Associates, 1984; Honma et al., 1999a), others positive results (McGregor et al., 1988). Positive results were found only at reduced RTG (relative total growth) values (McGregor et al., 1988). In one study, the incubation time was increased to 24 h to render the test system especially sensitive for the detection of clastogens and spindle poisons. In this study, the test substance was positive in one of two trials (Honma et al., 1999b).
In a cytogenetic assay with Chinese Hamster Ovary (CHO) cells, no significant increase in chromosomal aberrations was observed (Gulati et al., 1989). A further chromosomal aberration assay performed on Chinese hamster lung (CHL) cells was positive with metabolic activation only after a modified treatment of the cells (cells were treated for 6 h and then cultured in fresh medium for another 18 h) (Matsuoka et al., 1996). Increased chromosome aberrations without metabolic activation were only observed at cytotoxic concentrations (Matsuoka et al., 1996).
Gulati et al. (1989) reported a significant increase in SCE frequency at concentrations of 500 – 1000 mg/L induced only in the absence of S9 mix (no increase in the presence of Aroclor 1254-induced rat liver S9 mix). As these high test substance concentrations were cytostatic, increased SCE frequencies could only be detected after delayed harvest.
The test substance was tested for the induction of unscheduled DNA synthesis (UDS) in rat primary hepatocytes. Concentrations ranged from 0.005 - 0.4 μL/mL, the highest concentration being toxic. No increase in the mean nuclear grain count (as compared to controls) or in the incidence of cells undergoing repair was detected at any dose level (Microbiological Associates, 1984). However, another UDS test showed positive results (Selden et al., 1994).
Furthermore, in a transformation assay the test substance was evaluated as active (Matthews et al., 1993). Two umu assays were negative with and (weakly) positive without metabolic activation (Ono et al., 1991; Yasunaga et al., 2004). Positive results were also seen in a bacillus subtilis recombination assay (Matsui et al., 1989).
In vivo studies
In a mouse micronucleus assay, 496.8 mg/kg (= LD20 = MTD) test substance was administered i.p. to 5 male and 5 female CD-1 mice per group. Sampling time was 12, 24, 48 hrs post dosing. Significant increases in the number of micronucleated polychromatic erythrocytes (PCE) were not observed (Microbiological Associates, 1984). Likewise, in a study with CFLP mice (5 animals/dose/sex; gavage doses of 450, 900, 1800 mg/kg bw given in 2 equal parts (i.e. half of total dose) separated by an interval of 24 hrs) a negative result was obtained 6 hours after the last dosage of the test substance (Huntingdon, 1978). The test substance was also negative a bone marrow chromosomal abberrration test in mice (NTP, 1990).
In a DNA binding study, male and female F344 rats and B6C3F1 mice were administered 500 mg/kg [1,3,5-14C]-isophorone. Livers and kidneys (target organs in the NTP carcinogenicity study) were removed after 24 hrs. There was no binding of the test substance or its metabolites to DNA of these organs and it was concluded, that it is not likely that the tumors found in male F344 rats and B6C3F1 mice following long-term oral administration (Bucher et al., 1986) were caused by genotoxic effects (Thier et al., 1990).
In a Sex Linked Recessive Lethal (SLRL) test with Drosophila melanogaster, fruit flies were fed a diet containing 2000 ppm test substance. After 72 hrs of exposure, surviving males were mated. As feeding exposure was found to be non-mutagenic, 2 - 3 day old males were then injected with a 0.7 % NaCl solution containing 12,500 ppm test substance. 24 hrs post injection, males were mated. Again there was no indication of a mutagenic effect (Foureman et al., 1994).
Justification for classification or non-classification
The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. Some of the in vitro tests revealed inconclusive or positive results. However, all in vivo tests were clearly negative. Therefore, the substance is not considered to be classified for genotoxicity under Regulation (EC) No. 1272/2008, as amended for the 14th time in Regulation (EU) 2020/217.
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