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EC number: 948-047-4 | CAS number: -
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
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- Density
- Particle size distribution (Granulometry)
- Vapour pressure
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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
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- Toxicological Summary
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- Acute Toxicity
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- Carcinogenicity
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- Specific investigations
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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 reliable in vitro genotoxicity study (Ames test) a structurally similar substance was tested for mutagenicity in Salmonella typhimurium strains TA1535, TA1537, TA 1538, TA98 and TA100. Concentrations of up to 5000 μg/plate were tested. No evidence of mutagenic activity was seen at any concentration of the substance. It was concluded that the substance showed no evidence of mutagenic activity in this bacterial system under the test conditions employed, with and without a metabolic activation system.
In a reliable in vitro gene mutation study a structurally similar substance was tested for mutagenicity in mouse lymphoma L5178Y cells in the presence and absence of a metabolic activation system. Concentrations of up to333.0 µg/mL were tested. No evidence of mutagenic activity was seen at any concentration of the substance. It was concluded that the substance showed no evidence of mutagenic activity in mouse lymphoma L5178Y cells in the presence and absence of a metabolic activation system.
In a reliable in vitro chromosome aberration study, a structurally similar substance was tested for its ability to induce chromosomal aberrations in an in vitro mammalian cell system (V79 Chinese hamster lung cells), both in the presence and absence of exogenous metabolic activation. The substance was negative in the chromosome aberration test employing cultured human lymphocytes, either in the presence or absence of a metabolic activation system (rat liver S9).
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Remarks:
- Read-across to a K1 study therefore K2 is the maximum Klimisch score that can be assigned
- Justification for type of information:
- A read-across justification is provided in section 13 of the IUCLID file.
- Reason / purpose for cross-reference:
- read-across source
- Target gene:
- his operon
- Species / strain / cell type:
- S. typhimurium, other: TA98, TA100, TA1535, TA1537 and TA1538
- 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:
- First and second experiment: 8, 40, 200, 1000 and 5000 µg/plate with and without metabolic activation
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Tween 80; 1:1 (w/w); dilution to desired test concentrations with water
- Untreated negative controls:
- yes
- Remarks:
- untreated suspension
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Tween 80/bidist. water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: without S9: sodium azide (2 µg/plate) for TA100 and TA1535; 9-aminoacridine (80 µg/plate) for TA1537; 4-nitro-o-phenylendiamine (40 µg/plate) for TA98 and TA1538
- Untreated negative controls:
- yes
- Remarks:
- untreatred suspension
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Tween 80/bidist. water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: with S9: 2-aminoanthracene (2.5 µg/plate) for TA1535 and TA1537; (5 µg/plate) for TA98, TA100 and TA1538
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 h
NUMBER OF REPLICATIONS: 3 replications each in 2 independent experiments
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth - Evaluation criteria:
- A combination of the following criteria was considered as a positive result: the plate background of non-reverted bacteria did not show any growth reduction versus the respective negative controls; the spontaneous mutation rates of each tester strain per plate were within the characteristic spontaneous mutation range; as a rule, the positive controls showed mutation rates exceeding the control values of TA 100 at least by the factor 2.0 and those of the other tester strains at least by the factor 3.03; at more than one dose tested, the test substance caused at least a 2.0-fold increase in comparison with the negative controls in the tester strain TA 100. For the other tester strains, an increase in the mutation rate of more than 3.0 above the corresponding negative controls was considered positive.
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: noted at concentrations of 5000 µg/plate - Conclusions:
- The substance showed no evidence of mutagenic activity in this bacterial system under the test conditions employed, with and without a metabolic activation system.
- Executive summary:
In the in vitro genotoxicity study (Ames test) the substance was tested for mutagenicity in Salmonella typhimurium strains TA1535, TA1537, TA 1538, TA98 and TA100. Concentrations of up to 5000 μg/plate were tested. No evidence of mutagenic activity was seen at any concentration of the substance. It was concluded that the substance showed no evidence of mutagenic activity in this bacterial system under the test conditions employed, with and without a metabolic activation system..
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- Read-across to a K1 study therefore K2 is the maximum Klimisch score that can be assigned
- Justification for type of information:
- A read-across justification is provided in section 13 of the IUCLID file.
- Reason / purpose for cross-reference:
- read-across source
- Target gene:
- TK locus
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: RPMI 1640 Hepes buffered medium (Dutch modification) containing penicillin/streptomycin, sodium pyruvate and L-glutamin amd 10% (v/v) heat-inactivated horse serum (24 h exposure); for 3 h exposure only 5% (v/v) heat-inactivated horse serum were included. Selective medium consisted of the basic medium and 20% (v/v) heat-inactivated horse serum and 5 µg/mL trifluorothymidine (TFT).
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically "cleansed" against high spontaneous background: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital/β-naphtoflavone-induced rat liver S9
- Test concentrations with justification for top dose:
- Experiment 1:
With and without S9-mix: 0.1, 0.3, 1.0, 3.0, 10.0, 33.0, 100.0 and 333.0 µg/mL (3 h)
Experiment 2:
In the absence of S9-mix: 3.0, 10.0, 33.0, 100.0, 125.0, 140.0 and 175.0 µg/mL for 24h
In the presence of 12% (v/v) S9-mix: 0, 0.1, 0.3, 1.0, 3.0, 10.0, 33.0, 100.0 and 333.0 µg/mL for 3h - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: cyclophosphamide, 7.5 µg/mL, with S9; methylmethanesulfonate, 15 µg/mL and 5 µg/mL without S9, for 3 h and 24 h treatment, respectively
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 3 and 24 h
- Expression time (cells in growth medium): 48 h
SELECTION AGENT (mutation assays): 5 µg/mL trifluorothymidine
- Selection time (if incubation with a selection agent): 11-12 days
NUMBER OF REPLICATIONS: 2 replications each in 5 independent experiments
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency, relative total growth, relative survival, suspension growth, relative suspension growth, growth rate - Evaluation criteria:
- A mutation assay was considered acceptable if it met the following criteria:
a) The absolute cloning efficiency of the solvent controls is between 65 and 120%. An acceptable number of surviving cells (10E6) could be analysed for expression of the TK mutation.
b) The spontaneous mutation frequency in the solvent control is ≥ 50E-06 and ≤ 170E-06.
c) The growth rate (R) over the 2-day expression period for the negative controls should be between 8 and 32 (3 h treatment) and between 32 and 180 (24 h treatment).
d) The mutation frequency of MMS should not be below 500E-06, and fro CP not below 700E-06.
The global evaluation factor (GEF) has been identified by the IWTG as the mean of the negative/solvent mutation frequeny (MF) distribution plus one standard deviation. For the micro well version of the assay the GEF is 126.
A test substance is considered positive (mutagenic) in the mutation assay if it induces a MF of more than the MF of the controls + 126 in a dose-dependent manner. An observed increase should be biologically relevant and will be compared with the historical control data range.
A test substance is considered equivocal (questionable) in the mutation assay if no clear conclusion for positive or negative result can be made after an additional confirmation study.
A test substance is considered negative (not mutagenic) in the mutation assay if: a) None of the tested concentrations reaches a mutation frequency of the MF of the controls + 126. b) The results are confirmed in an independently repeated test. - Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility and precipitation: The test substance precipitated in the exposure medium at concentrations of 100 µg/mL and above. The test substance was tested beyond the limit of the solubility to obtain adequate cytotoxicity data, the concentration used as the highest test substance concentration for the dose range ending test was 333 µg/mL.
RANGE-FINDING/SCREENING STUDIES:
In the absence of S9-mix, the relative suspension growth was 66% at the test substance concentration of 333 µg/mL compared to the relative suspension growth of the solvent control. In the presence of S9-mix, no toxicity in the relative suspension growth was observed up to and including the highest test substance concentration of 333 µg/mL compared to the suspension growth of the solvent control.
COMPARISON WITH HISTORICAL CONTROL DATA: The spontaneous mutation frequencies in the solvent treated control cultures were within the ranges of the historical controls. - Conclusions:
- It was concluded that the substance showed no evidence of mutagenic activity in mouse lymphoma L5178Y cells in the presence and absence of a metabolic activation system.
- Executive summary:
In the in vitro gene mutation study the substance was tested for mutagenicity in mouse lymphoma L5178Y cells in the presence and absence of a metabolic activation system. Concentrations of up to333.0 µg/mLwere tested. No evidence of mutagenic activity was seen at any concentration of the substance. It was concluded that the substance showed no evidence of mutagenic activity inmouse lymphoma L5178Y cells in the presence and absence of a metabolic activation system.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study without detailed documentation
- Remarks:
- Read-across to K2 study therefore K2 is the maximum Klimisch value.
- Justification for type of information:
- Read-across approach - see read-across justification in section 13.
- Reason / purpose for cross-reference:
- read-across source
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Specific details on test material used for the study:
- Batch: 704025
Storage: dark, without air humidity, under exhaust hood, at room temperature - Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- Cloned V79 cells with a modal chromosome number of 22 and a cell cycle length of approx. 16.5 h were used.
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 fraction was obtained from the livers of 8 - 10 weeks old male Sprague Dawley rats which received a single i.p. injection of 500 mg/kg b.w. Arochlor 1254 in corn oil 5 days prior to the preparation of the S9 fraction.
- Test concentrations with justification for top dose:
- Ten different concentrations in the range of 1 to 100 µg/ml were tested. The doses were selected based on the solubility of the substance in ethanol.
- Vehicle / solvent:
- Ethanol
- Untreated negative controls:
- yes
- Remarks:
- MEM4 medium containing 1 % ethanol served as the negative control
- Negative solvent / vehicle controls:
- no
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- mitomycin C
- Rationale for test conditions:
- As per OECD Test Guidelines
- Evaluation criteria:
- The test chemical is to be considered clastogenic in this assay if:
1. it induces chromosomal aberrations (excl. gaps) in a statistically significant manner in one or more concentrations.
2. the induced proportion of aberrant cells at such test substance concentrations exceeds the normal range of the test system (i.e. » 5%).
3. positive results can be varified in an independent experiment. - Statistics:
- The proportion of cells that was treated with the test substance and harboured structural aberrations (excl. gaps) was compared with the corresponding proportion of the negative controls in the Chi-square test. Probability values of p < 0.05 were accepted as statistically significant.
- Key result
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- not applicable
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- The frequency of polyploid cells in both parts of the experiment was within the expected range (<10 %).
- Conclusions:
- The substance was negative in the chromosome aberration test employing cultured human lymphocytes, either in the presence or absence of a metabolic activation system (rat liver S9).
- Executive summary:
The substance was tested for its ability to induce chromosomal aberrations in an in vitro mammalian cell system (V79 Chinese hamster lung cells). V79 cells were exposed to the substance both in the presence and absence of exogenous metabolic activation by Arochlor 1254 induced rat liver S9. Cells were arrested in metaphase by 2 h treatment with Colcemid 16 and 26 h after the start of exposure. After hypotonic treatment they were fixed and Giemsa stained. The mitotic indices of representative cultures, as a measure for cytotoxicity, were determined and metaphase cells were analysed for the presence of chromosomal aberrations. For each experimental point, at least duplicate cultures (100 metaphases per culture) were evaluated. To demonstrate the sensitivity of the test system, Mitomycin C (without S9 mix) and Cyclophosphamide (with S9 mix) were used as positive controls.
The experiments with and without S9 mix revealed a systematic influence of the test compound which led to a reduction in the mitotic index, which was used as a measure of cytotoxicity. In the chromosome aberration studies with metabolic activation cells were treated with 3 concentrations ranging from
10 to 100 µg/ml. In the corresponding experiments without metabolic activation 3 concentrations ranging from 10 to 80 µg/ml were applied. In both experiments, at the 18 h as well as the 28 h sampling time, the substance did not induce significant increases in the proportion of cells with chromosomal
aberrations (excluding gaps). The positive controls, Mitomycin C and Cyclophosphamide, did induce chromosomal aberrations, thus demonstrating the sensitivity of the test system against clastogenic agents. The substance was negative in the chromosome aberration test employing cultured human lymphocytes, either in the presence or absence of a metabolic activation system (rat liver S9).
Referenceopen allclose all
Table 1: Maximum revertants of both experimental series
|
Maximum number of revertants (dose level in µg/mL) |
|||||
solvent control |
positive control |
test substance |
||||
Strain |
With S9 |
Without S9 |
With S9 |
Without S9 |
With S9 |
Without S9 |
TA 98 |
45.0 |
25 |
1218 |
643 |
46.7 (1000) |
24.3 (1000) |
TA 100 |
132.7 |
111 |
936.7 |
318.0 |
122.3 (1000) |
115.0 (1000) |
TA 1535 |
19.0 |
12.3 |
274.0 |
384.0 |
15.0 (1000) |
13.0 (200) |
TA 1537 |
11.3 |
9.3 |
124.7 |
955.3 |
13.3 (200) |
11.3 (1000) |
TA 1538 |
18.7 |
9.0 |
1499.7 |
1678.0 |
24.3 (200) |
10.7 (40) |
Table 1: Cytotoxic and mutagenic responses
Treatment |
Concentration [µg/mL] |
Cloning efficiency [%] |
Relative total growth [%] |
Mutation frequency x 10-6
|
||
total |
small colonies |
large colonies |
||||
3 h treatment without S9-mix |
||||||
Solvent control (mean) |
-- |
86 |
100 |
61 |
42 |
19 |
Test substance |
0.1 |
101 |
114 |
56 |
29 |
25 |
0.3 |
101 |
108 |
52 |
28 |
23 |
|
1.0 |
118 |
140 |
66 |
48 |
16 |
|
3.0 |
91 |
109 |
63 |
46 |
15 |
|
10.0 |
95 |
108 |
66 |
32 |
33 |
|
33.0 |
97 |
104 |
76 |
46 |
28 |
|
100.0* |
91 |
97 |
81 |
32 |
46 |
|
333.0* |
102 |
77 |
64 |
48 |
14 |
|
MMS |
15 |
60 |
45 |
685 |
521 |
118 |
3 h treatment with 8% (v/v) S9-mix |
||||||
Solvent control (mean) |
-- |
91 |
100 |
67 |
36 |
29 |
Test substance |
0.1 |
108 |
111 |
74 |
47 |
25 |
0.3 |
89 |
94 |
71 |
45 |
24 |
|
1.0 |
108 |
113 |
64 |
42 |
20 |
|
3.0 |
98 |
107 |
78 |
53 |
23 |
|
10.0 |
95 |
100 |
87 |
54 |
29 |
|
33.0 |
108 |
96 |
55 |
32 |
22 |
|
100.0* |
98 |
89 |
83 |
60 |
21 |
|
333.0* |
94 |
92 |
63 |
23 |
39 |
|
CP |
7.5 |
60 |
32 |
1074 |
829 |
144 |
24 h treatment without S9-mix |
||||||
Solvent control (mean) |
-- |
115 |
100 |
51 |
26 |
23 |
Test substance |
3 |
135 |
92 |
58 |
42 |
14 |
10 |
137 |
109 |
51 |
39 |
11 |
|
33 |
133 |
85 |
71 |
32 |
35 |
|
100* |
110 |
30 |
100 |
35 |
60 |
|
125* |
118 |
31 |
70 |
31 |
36 |
|
140* |
118 |
20 |
103 |
51 |
47 |
|
175* |
102 |
9 |
134 |
53 |
72 |
|
MMS |
5 |
101 |
73 |
865 |
463 |
233 |
3 h treatment with 12% (v/v) S9-mix |
||||||
Solvent control (mean) |
-- |
109 |
100 |
72 |
47 |
23 |
Test substance |
0.1 |
110 |
100 |
73 |
49 |
21 |
0.3 |
123 |
117 |
72 |
47 |
23 |
|
1.0 |
104 |
105 |
82 |
52 |
27 |
|
3.0 |
97 |
104 |
94 |
62 |
29 |
|
10.0 |
115 |
126 |
87 |
57 |
26 |
|
33.0 |
107 |
108 |
85 |
59 |
23 |
|
100.0* |
131 |
123 |
80 |
51 |
26 |
|
333.0* |
97 |
83 |
92 |
64 |
24 |
|
CP |
7.5 |
70 |
59 |
979 |
621 |
221 |
*precipitation of test substance in the exposure medium
MMS = methylmethanesulfonate
CP = cyclophosphamide
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Justification for classification or non-classification
Based on the findings of reliable in vitro genotoxicity studies conducted on structurally similar substances, classification of the substance is not justified.
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.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.