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EC number: 475-290-9 | CAS number: 39537-23-0
- 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
Gene mutation in bacteria (OECD 471, Ames test): negative in S. typhimurium strains TA98, TA100, TA1535 and TA1537 with and without metabolic activation
Gene mutation in bacteria (OECD 471, Ames test): negative in S. typhimurium strains TA98, TA100, TA1535 and TA1537 and in E. coli strain WP2uvrA with and without metabolic activation
Cytogenicity in mammalian cells (similar to OECD 473, Chromosome aberration test): negative in Chinese hamster lung cells with and without metabolic activation
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 1994-02-10 to 1994-02-18
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- 26 May 1983
- Deviations:
- yes
- Remarks:
- no S. typhimurium T102 or E. coli strain tested; only 2-aminoanthracene used as positive control with S9 mix
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his operon
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Metabolic activation system:
- Cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254.
- Test concentrations with justification for top dose:
- 0, 62, 185, 556, 1667, 5000 µg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: water
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- 2-nitrofluorene
- sodium azide
- other: 2-Aminoanthracene
- Details on test system and experimental conditions:
- NUMBER OF REPLICATIONS:
- Number of cultures per concentration: triplicate
- Number of independent experiments: 2
METHOD OF TREATMENT/ EXPOSURE:
- Test substance added in agar (plate incorporation, both experiments)
TREATMENT AND HARVEST SCHEDULE:
- Exposure duration/duration of treatment: 72 h at 37 °C
METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method: Inspection of the bacterial background lawn - Evaluation criteria:
- A positive response is a two-fold or greater increase in the mean number of revertant colonies appearing in the test plates over and above the background spontaneous reversion rate observed with the vehicle, together with evidence of a dose-response.
- Statistics:
- Mean values and standard deviation were calculated.
- Species / strain:
- S. typhimurium, other: TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: at 5000 µg/plate slightly less dense background lawn of bacterial growth than in concomitant control plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation and time of the determination: No precipitation of the test substance was observed.
RANGE-FINDING/SCREENING STUDIES:
A preliminary test to assess the toxicity of the test substance to the bacteria was not performed as no toxicity was expected. The cytogenicity test was incorporated into the first mutagenicity assay. - Conclusions:
- The test substance shows no mutagenic activity in Salmonella typhimurium TA1535, TA 1537, TA98 or TA 100, either in the absence or in the presence of the S9-mix.
- 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:
- comparable to guideline study with acceptable restrictions
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- Not applicable
- Species / strain / cell type:
- Chinese hamster lung (CHL/IU)
- Details on mammalian cell type (if applicable):
- For cell lines:
- Cell cycle length: 17 h
MEDIA USED
- Type and composition of media, CO2 concentration, humidity level, temperature: cells were grown in Eagle’s Minimal Essential Media with HEPES buffer and Earle’s Salts, supplemented with 10% foetal bovine serum and antibiotics and incubated at 37 °C with 5% CO2 in air - Metabolic activation:
- with and without
- Metabolic activation system:
- Type and composition of metabolic activation system:
- source of S9: male Sprague–Dawley rats (approx. 250 g) induced with 3 consecutive daily doses of 80 mg phenobarbitone/kg bw and 100 mg beta-naphthoflavone/kg bw
- quality controls of S9: metabolic capability tested - Test concentrations with justification for top dose:
- Preliminary toxicity test: 34 to 2180 µg/mL
First and second experiment: 0 to 2180 µg/mL - Vehicle / solvent:
- - Vehicle/solvent used: sterile distilled water
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- mitomycin C
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION first experiment
- Exposure duration: 6 h with and without S9-mix
- Recovery phase: 18 h
- Fixation time (start of exposure up to fixation or harvest of cells): 24 h
DURATION second experiment
- Exposure duration: 24 h or 48 h without S9-mix
- Fixation time (start of exposure up to fixation or harvest of cells): 24 h or 48 h
SPINDLE INHIBITOR (cytogenetic assays): 0.1 µg/ml Colcemid two hours prior to the end of the incubation
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS:
- each concentration in duplicate
NUMBER OF CELLS EVALUATED:
- the first 100 consecutive well-spread metaphases from each culture to a total of 200 cells per concentration
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index calculated from 1000 cells counted & metaphases recorded
OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreplication: yes - Key result
- Species / strain:
- other: Chinese Hamster Lung (CHL) 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
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Data on pH: no effect on pH when dosed in cell culture
- Data on osmolality: no effect on osmolality when dosed in cell culture
RANGE-FINDING/SCREENING STUDIES:
The test item at concentrations ranging from 34 to 2180 µg/mL was incubated with cell cultures for 24 and 48 h without S9-mix, and for 6 h both with and without S9-mix. The highest concentration analysed was selected based on the recommendation limit dose. - Conclusions:
- Under the test conditions applied, the test substance does not induce chromosome aberrations in cultured mammalian somatic cells, in the presence and absence of a metabolic activation system.
- 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:
- comparable to guideline study with acceptable restrictions
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his operon for S. typhimurium strains and trp operon for E. coli strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Details on mammalian cell type (if applicable):
- CELLS USED
- Type and source of cells: University of California, Berkeley, USA - Species / strain / cell type:
- E. coli WP2 uvr A
- Details on mammalian cell type (if applicable):
- CELLS USED
- Type and source of cells: British Industrial Biological Research Association, UK - Metabolic activation:
- with and without
- Metabolic activation system:
- Type and composition of metabolic activation system:
- source of S9 : male Sprague–Dawley rats (approx. 250 g) induced with 3 consecutive daily doses of 80 mg phenobarbitone/kg bw and 100 mg beta-naphthoflavone/kg bw
- quality controls of S9: metabolic capability tested - Test concentrations with justification for top dose:
- Range finding assay: 0, 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500, 5000 µg/plate
Main study and repeat study: 50, 150, 500, 1500, 5000 µg/plate - Vehicle / solvent:
- - Vehicle/solvent used: sterile distilled water
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 9-aminoacridine
- N-ethyl-N-nitro-N-nitrosoguanidine
- benzo(a)pyrene
- other: 2-aminoanthracene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 h at 37 °C
NUMBER OF REPLICATIONS: triplicates each in 2 independent experiments
METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method: background growth inhibition - Species / strain:
- S. typhimurium TA 1535
- 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
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- 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
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 98
- 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
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- 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
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A
- 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
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- RANGE-FINDING/SCREENING STUDIES:
In the range finding test, the substance was tested up to 5000 µg/plate in the absence and presence of S9 mix in S. typhimurium TA100 and E. coli WP2uvrA. No reduction of the bacterial lawn and no biologically relevant decrease in the number of revertants were observed. The highest concentration analysed was selected based on the recommended limit dose in OECD guideline 471. - Conclusions:
- Under the conditions of the test the test substance is considered to be not mutagenic in S. typhimurium TA1537, TA1535, TA100, and TA98 as well as E. coli WP2uvrA, in the presence and absence of a metabolic activation system.
Referenceopen allclose all
Table 1: Detailed results of the first experiment
Dose µg/plate |
|
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
||||
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
||
0 |
Mean SD |
22 25 18
22 4 |
18 14 17
16 2 |
16 13 16
15 2 |
20 15 12
16 4 |
31 30 33
31 2 |
47 45 52
48 4 |
163 197 159
173 21 |
177 167 181
175 7 |
62 |
Mean SD |
19 19 27
22 5 |
13 11 23
16 6 |
14 12 17
14 3 |
17 22 12
17 5 |
44 40 40
41 2 |
52 70 46
56 12 |
211 175 176
187 21 |
174 211 190
192 19 |
185 |
Mean SD |
30 22 25
26 4 |
24 15 19
19 5 |
28 24 19
24 5 |
19 19 13
17 3 |
51 36 37
41 8 |
62 56 47
55 8 |
170 172 195
179 14 |
175 177 170
174 4 |
556 |
Mean SD |
20 23 25
23 3 |
23 20 26
23 3 |
14 12 22
16 5 |
23 20 19
21 2 |
33 38 51
41 9 |
53 53 58
55 3 |
196 174 183
184 11 |
178 187 174
180 7 |
1667 |
Mean SD |
30 26 19
25 6 |
23 15 18
19 4 |
13 15 12
13 2 |
19 13 14
15 3 |
34 38 39
37 3 |
52 58 39
50 10 |
175 192 181
183 9 |
193 178 172
181 11 |
5000 |
Mean SD |
12 18 23
18 6 |
14 12 15
14 2 |
11 14 16
14 3 |
16 22 15
18 4 |
40 37 38
38 2 |
37 59 45
47 11 |
107 131 126
121 13 |
177 208 164
183 23 |
Positive Control |
Mean SD |
540 502 502
515 22 |
473 487 472
477 8 |
1085 904 958
982 93 |
195 148 193
179 27 |
452 462 526
480 40 |
1367 1294 1335
1332 37 |
604 608 619
610 8 |
1663 1782 1689
1711 63 |
SD: Standard deviation
Table 2: Detailed results of the second experiment
Dose µg/plate |
|
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
||||
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
||
0 |
Mean SD |
28 34 25
29 5 |
17 20 16
18 2 |
25 23 20
23 3 |
19 19 30
23 6 |
30 37 55
41 13 |
57 63 53
58 5 |
162 186 157
168 16 |
187 79 186
184 4 |
62 |
Mean SD |
17 17 24
19 4 |
15 22 19
19 4 |
18 16 22
19 3 |
16 17 25
19 5 |
36 38 42
39 3 |
38 58 48
48 10 |
163 163 174
167 6 |
165 140 173
159 17 |
185 |
Mean SD |
29 11 17
19 9 |
16 16 25
19 5 |
25 20 24
23 3 |
16 20 14
17 3 |
30 33 34
32 2 |
60 50 51
54 6 |
165 177 184
175 10 |
150 165 159
158 8 |
556 |
Mean SD |
22 27 29
26 4 |
20 22 24
22 2 |
13 14 12
13 1 |
16 16 17
16 1 |
55 40 42
46 8 |
57 58 61
59 2 |
174 207 167
183 21 |
189 161 164
171 15 |
1667 |
Mean SD |
35 40 24
33 8 |
27 15 22
21 6 |
27 23 25
25 2 |
16 14 22
17 4 |
47 48 44
46 2 |
59 68 40
56 14 |
173 188 175
179 8 |
173 164 189
175 13 |
5000 |
Mean SD |
25 20 20
22 3 |
17 16 19
17 2 |
20 17 27
21 5 |
22 22 15
20 4 |
45 46 42
44 2 |
67 53 68
63 8 |
143 117 127
129 13 |
189 170 192
184 12 |
Positive Control |
Mean SD |
499 445 506
483 33 |
445 412 424
427 17 |
1408 1208 1380
1332 108 |
185 145 188
173 24 |
493 524 495
504 17 |
1528 1446 1539
1504 51 |
633 600 616
616 17 |
1754 1743 1687
1728 36 |
SD: Standard deviation
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Gene mutation in bacteria
An Ames test was described in a publication (Oda_et_al_2008_Ames), which was conducted similarly to OECD guideline 471 and compliant with GLP. Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 and Escherichia coli strain WP2uvrA were exposed to the test substance, vehicle and positive controls in the presence and absence of metabolic activation (S9 mix). A range-finding assay with concentrations in the range of 0.15 to 5000 µg/plate was conducted prior to the main mutagenicity test, with and without S9 mix. Based on the results of this pre-test, test substance concentrations in the range of 50 to 5000 µg/plate were chosen for the mutagenicity assay. In two independent experiments with and without metabolic activation, the bacteria were exposed to the test substance or corresponding controls in agar (plate-incorporation test). After 48 h of exposure, the bacterial background lawn was inspected and the number of revertant colonies was determined.
There was no cytotoxicity up to the highest concentration tested in the presence and in the absence of S9 mix. Treatment with the test did not increase the number of revertant colonies at any dose level for any of the bacteria strains tested, with or without S9 mix. The vehicle and positive control showed the expected results, indicating that the test system was sensitive and valid.
Based on the experimental results, the test item is considered not mutagenic in bacteria, with and without metabolic activation.
The test substance was also investigated for its potential to cause gene mutation in bacteria in an Ames test which was performed according to OECD guideline 471 and in compliance with GLP (93-0104-DNT). However, compared to the current OECD guideline 471, there are deviations: only four bacterial strains were tested and only 2-aminoanthracene was used as positive control substance in the presence of a metabolic activation system.
Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 were exposed to the test substance, vehicle (water) or positive controls in the presence and absence of metabolic activation (S9 mix). In two independent experiments, test substance concentrations in the range of 62 to 5000 µg/plate were added to the agar (plate-incorporation method) with and without S9 mix. After 72 h of exposure at 37 °C, the bacterial background lawn was inspected and the number of revertant colonies were counted for each strain.
There was no precipitation observed up to and including the highest concentration tested. At 5000 µg/plate in the absence of metabolic activation, slight indications of cytotoxicity were noted. Treatment with the test item did not increase the number of revertant colonies for any tester strain, neither in the presence, nor in the absence of metabolic activation. The mean number of revertant colonies was comparable to the spontaneous reversion rate of vehicle controls. The positive control induced a strong increase in the number of revertant colonies, demonstrating the functionality of the test. Under the conditions of the experimental study, there is no indication of mutagenicity in bacteria, in the presence and in the absence of metabolic activation.
In vitro cytogenicity in mammalian cells
The Genetic toxicity in vitro was tested in a chromosomal aberration assay that was published (Oda_et_al_2008_CA). The test was performed according to OECD guideline 473 and compliant with GLP. In two independent experiments, duplicate cultures of Chinese hamster lung (CHL) cells were exposed to the test substance in the presence and absence of metabolic activation (S9 mix). The cells were exposed to test substance concentrations of 1.090, 1.635 and 2.180 µg/mL for 6 h in the presence and absence of S9 mix (first experiment, sampling 24 h after start of exposure) and for 24 or 48 h in the absence of S9 mix (second experiment, sampling at the end of exposure). A total of 200 metaphases for scored for each condition and the mitotic index (MI) indicating cytotoxicity was assessed for each condition.
There was no cytotoxicity up the highest concentration in the presence or in the absence of metabolic activation. Treatment with the test substance did not induce the mean incidence of chromosomal aberrations at any dose level, in the presence and in the absence of S9 mix. Aberrant cell frequencies induced by the positive control substances cyclophosphamide (with metabolic activation) and mitomycin C (without metabolic activation) demonstrated clastogenic activity and confirmed the validity of the test system.
Based on the results of the study the test substance does not induce chromosomal aberrations in mammalian cells.
Conclusion
In conclusion, the available experimental data on gene mutation in bacteria and chromosome aberration in mammalian cells in vitro do not indicate that the test substance is mutagenic or clastogenic in vitro, in the presence and in the absence of metabolic activation.
Justification for classification or non-classification
The available data on genetic toxicity in vitro do not meet the criteria for classification according to Regulation (EC) No. 1272/2008 (CLP), and are therefore conclusive but not sufficient for classification.
No classification for genetic toxicity is warranted according to the criteria of the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations.
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