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EC number: 203-756-1 | CAS number: 110-31-6
- 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
In vitro:
Gene mutation in bacteria (Reverse Mutation Test, similar to OECD 471):
negative, based on read-across from Amides, C16-C18 (even),
N,N’-ethylenebis
Cytogenicity in mammalian cells (Chromosomal Aberration, similar to OECD
473): negative, based on read-across from Amides, C16-C18 (even),
N,N’-ethylenebis
Gene mutation in mammalian cells (Mouse Lymphoma Assay, OECD 476):
negative, based on read-across from Amides, C16-C18 (even),
N,N’-ethylenebis
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- migrated 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:
- other: Comparable to guideline study with acceptable restrictions. Strain to detect cross-linking mutagens missing.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- strain to detect cross-linking mutagens missing
- GLP compliance:
- no
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his operon
- Species / strain / cell type:
- other: S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100
- Species / strain / cell type:
- other: Saccharomyces cerevisiae D4
- Metabolic activation:
- with and without
- Metabolic activation system:
- Liver S9 fraction of Aroclor 1254-pretreated rats
- Test concentrations with justification for top dose:
- 0.1, 1.0, 10.0, 100.0, 500.0 and 1000.0 µg/plate without metabolic activation
0.1, 1.0, 10.0, 100.0 and 500.0 µg/plate with metabolic activation - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: water, dimethylsulfoxid (DMSO)
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Dimethylsulfoxide (DMSO)
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- without metabolic activation
- Positive control substance:
- 2-nitrofluorene
- ethylmethanesulphonate
- other: quinacrine mustard
- Remarks:
- ethyl methanesulphonate (-S9; 10 µL/plate, TA1535, TA100, D4); quinacrine mustard (-S9; 10 µg/plate, TA1537); 2-nitrofluorene (-S9; 10 µg/plate, TA1538, TA98); Migrated to IUCLID6: 10 µL/plate
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- N-dimethylnitrosamine
- other: 2-anthramine
- Remarks:
- 2-anthramine (+S9; 2.5 µg/plate, TA1535, TA1537, TA1538, TA98, TA 100); N-dimethylnitrosamine (+S9; 100 µmol/plate, D4)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 h for TA 1535, TA 1537, TA 1538, TA 98, TA 100; 3-5 days for D4
DETERMINATION OF CYTOTOXICITY
- Method: other: direct revertant colony counts
- Evaluation criteria:
- Strains TA 1535, TA 1537, and TA 1538: if the solvent control value is within the normal range, a chemical that produces a positive dose-response over three concentrations with the lowest increase equal to twice the solvent control value is considered to be mutagenic.
Strains TA 98, TA 100, and D4: if the solvent control value is within the normal range, a chemical that produces a positive dose response over three concentrations with the highest increase equal to twice the solvent control value for TA 100 and 2-3 times the solvent control value for strains TA 98 and D4 is considered to be mutagenic. For these strains, the dose-response increase should start at approximately the solvent control value.
If a chemical produces a response in a single test that cannot be reproduced in one or more additional runs, the initial positive test data lose significance. - Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True 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:
- cytotoxicity
- Remarks:
- slightly toxic at 500 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True 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
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True 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
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True 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
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- Saccharomyces cerevisiae
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- ADDITIONAL INFORMATION ON CYTOTOXICITY: the compound was slightly toxic to the strain T-1537 at 500 µg per plate. TA-98 was repeated at 100, 500 and 1000 µg because of the increased number of revertants at 500 µg over the background level. The repeat test was negative.
- Conclusions:
- Interpretation of results:
negative - Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- 10 Mar – 27 Apr 2010
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- GLP - Guideline study, tested with the source substance N,N’-Ethylenebis(stearamide). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- thymidine kinase (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-glutamine + 10% (v/v) heat-inactivated horse serum (24-hour exposure); for 3-hour exposure only 5% (v/v) heat-inactivated horse serum were included. Selective medium consisted of the basic medium + 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 8% (v/v) S9-mix: 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.8 and 1.0 µg/mL for 3h
Experiment 2:
Without S9-mix: 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.8 and 1.0 µg/mL for 24h
With 12% (v/v) S9-mix: 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.8 and 1.0 µg/mL for 3h - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: hexane
- Justification for choice of solvent/vehicle: solubility/ability to suspend - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- hexane
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: cyclophosphamide
- Remarks:
- with S9-mix: 7.5 µg/mL
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- hexane
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: methylmethanesulfonate
- Remarks:
- without S9-mix: 15 µg/mL and 5 µg/mL 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, respectively
- Expression time (cells in growth medium): 2 days
- Selection time (if incubation with a selection agent): 11-12 days
SELECTION AGENT (mutation assays): 5 µg/mL trifluorothymidine (TFT) (Sigma)
NUMBER OF REPLICATIONS: 5 exposure plates
NUMBER OF CELLS EVALUATED: not applicable, number of mutants per well counted; 2000 cells/well inserted, total sum of mutants given
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 (10E+6) could be analysed for expression of the TK mutation.
b) The spontaneous mutation frequency in the solvent control is ≥ 50 x 10E-6 and ≤ 170 x 10E-6.
c) The growth rate (R) over the 2-day expression period for the negative controls should be between 8 and 32 (3 hours treatment) and between 32 and 180 (24 hours treatment).
d) The mutation frequency of MMS should not be below 500 x 10E-6, and fro CP not below 700 x 10E-6.
The global evaluation factor (GEF) has been identified by the IWTG as the mean of the negative/solvent mutation frequency (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: a slight precipitate of the test substance in the exposure medium was observed after 3 hours treatment at a concentration of 0.8 µg/mL and severe precipitate was observed at concentrations of 2.4 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 test was 24 µg/mL
RANGE-FINDING/SCREENING STUDIES:
Both in the absence and presence of S9-mix, no toxicity in the relative suspension growth was observed up to and including the highest test substance concentration of 24 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:
- Interpretation of results:
negative - Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- migrated 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:
- other: see 'Remark'
- Remarks:
- GLP - Guideline study, tested with the source substance N,N’-Ethylenebis(stearamide). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- THE DEPARTMENT OF HEALTH OF THE GOVERNMENT OF THE UNITED KINGDOM
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- Not applicable
- Species / strain / cell type:
- other: CHL/IU (Chinese Hamster Lung cell line)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Eagles Mimimal Essential Media with 10% Foetal Bovine Serum
- Properly maintained: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbitone/β-Naphtoflavone-induced rat liver S9
- Test concentrations with justification for top dose:
- 6(18)-hour without S9: 0, 4.89, 9.77, 19.53, 39.06, 78.13, 156.25 µg/mL
6(18)-hour with S9: 0, 4.89, 9.77, 19.53, 39.06, 78.13, 156.25 µg/mL
24-hour without S9: 0, 2.44, 4.89, 9.77, 19.53, 39.06, 78.13 µg/mL
Concentrations selected for metaphase analysis, highest concentration was lowest precipitating concentration:
6(18)-hour without S9: 0, 19.53, 39.06, 78.13 µg/mL
6(18)-hour with S9: 0, 19.53, 39.06, 78.13 µg/mL
24-hour without S9: 0, 9.77, 19.53, 39.06 µg/mL - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: mitomycin C
- Remarks:
- without S9-mix: 0.1 µg/mL for 6 h and 0.05 µg/mL for 24 h treatment
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- other: cyclophosphamide
- Remarks:
- with S9-mix: 5 µg/mL for 6 h treatment Migrated to IUCLID6: 5 µg/mL for 6h treatment
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 6 and 24h
- Expression time (cells in growth medium): 18 h after 6 h treatment
SPINDLE INHIBITOR (cytogenetic assays): colcemid 0.1 µg/mL
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 200 (100 per plate)
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index
OTHER EXAMINATIONS:
- Determination of polyploidy: yes - Evaluation criteria:
- Cells with 38 or more chromosomes were classified as polyploid cells and the % incidence of polyploid cells reported. Endoreduplicated cells were recorded separately and are included in the polyploid cell total number. If there was a dose-related increase in endoreduplicated cells then they are reported separately. The percentage of cells showing structural chromosome aberrations (breaks and exchanges) was calculated and reported. The number of gap-type aberrations was recorded and reported.
- Statistics:
- The frequency of cells with aberrations excluding gaps and the frequency of polyploid cells was compared, where necessary with the concurrent vehicle control value using Fisher‘s Exact test.
- Key result
- Species / strain:
- other: CHL/IU (Chinese Hamster Lung cell line)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: a precipitate of the test material was seen at and above 78.13 µg/mL in both pulse exposure groups and at and above 39.06 µg/mL in the 24 hours continuous exposure group
RANGE-FINDING/SCREENING STUDIES: In all cases the test material showed no evidence of cell toxicity. The dose selection for the main experiments was based on the lowest precipitating dose level, which was 78.13 µg/mL for both short term exposu.re groups and 39.06 µg/mL for the continuous exposure group. An additional dose above the lowest precipitating dose was included for all exposures. - Conclusions:
- Interpretation of results:
negative
Referenceopen allclose all
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 hours treatment without S9-mix |
||||||
Solvent control (mean) |
-- |
99 |
100 |
114 |
62 |
46 |
Test substance |
0 |
116 |
126 |
110 |
59 |
43 |
0.1 |
123 |
126 |
99 |
53 |
40 |
|
0.2 |
113 |
111 |
110 |
63 |
41 |
|
0.3 |
110 |
124 |
118 |
67 |
44 |
|
0.4 |
113 |
139 |
128 |
60 |
59 |
|
0.5 |
110 |
154 |
118 |
63 |
48 |
|
0.6* |
108 |
125 |
126 |
71 |
48 |
|
0.8* |
118 |
125 |
105 |
52 |
47 |
|
1.0* |
115 |
125 |
119 |
67 |
45 |
|
MMS |
15 |
58 |
55 |
1227 |
675 |
354 |
3 hours treatment with 8% (v/v) S9-mix |
||||||
Solvent control (mean) |
-- |
106 |
100 |
102 |
55 |
42 |
Test substance |
0 |
105 |
102 |
80 |
38 |
38 |
0.1 |
94 |
89 |
119 |
66 |
46 |
|
0.2 |
105 |
55 |
89 |
51 |
34 |
|
0.3 |
93 |
76 |
113 |
64 |
43 |
|
0.4 |
120 |
81 |
93 |
47 |
41 |
|
0.5 |
121 |
127 |
97 |
55 |
37 |
|
0.6* |
101 |
86 |
87 |
53 |
30 |
|
0.8* |
97 |
94 |
72 |
44 |
25 |
|
1.0* |
121 |
112 |
79 |
45 |
30 |
|
CP |
7.5 |
47 |
23 |
1431 |
876 |
351 |
24 hours treatment without S9-mix |
||||||
Solvent control (mean) |
-- |
93 |
100 |
58 |
29 |
27 |
Test substance |
0 |
86 |
138 |
66 |
34 |
31 |
0.1 |
72 |
108 |
77 |
43 |
32 |
|
0.2 |
79 |
121 |
78 |
38 |
38 |
|
0.3 |
98 |
150 |
47 |
26 |
19 |
|
0.4 |
90 |
151 |
55 |
27 |
26 |
|
0.5 |
81 |
133 |
69 |
37 |
30 |
|
0.6* |
80 |
125 |
67 |
35 |
31 |
|
0.8* |
90 |
143 |
61 |
37 |
22 |
|
1.0* |
88 |
131 |
63 |
34 |
27 |
|
MMS |
5 |
66 |
81 |
629 |
357 |
211 |
3 hours treatment with 12% (v/v) S9-mix |
||||||
Solvent control (mean) |
-- |
106 |
100 |
82 |
50 |
29 |
Test substance |
0 |
85 |
294 |
83 |
52 |
28 |
0.1 |
107 |
58 |
109 |
48 |
55 |
|
0.2 |
118 |
60 |
77 |
42 |
32 |
|
0.3 |
91 |
161 |
88 |
50 |
34 |
|
0.4 |
85 |
127 |
86 |
55 |
28 |
|
0.5 |
101 |
146 |
74 |
49 |
23 |
|
0.6* |
86 |
42 |
103 |
66 |
32 |
|
0.8* |
93 |
155 |
67 |
40 |
25 |
|
1.0* |
62 |
66 |
130 |
55 |
70 |
|
CP |
7.5 |
37 |
43 |
1905 |
1113 |
500 |
*precipitation of test substance in the exposure medium
The test substance did not induce a significant increase in mutation frequency in the absence or presence of metabolic activation in the first experiment. This result was confirmed in an independent experiment with a different concentration of the metabolic activation system or a longer exposure time without activation.
Table 1: Chromosome aberrations and cell viability
Test item |
Concentration |
Cell viability |
Mean |
Aberrant cells in % |
|
|
in µg/mL |
in % |
mitotic index |
Numerical |
Structural |
Exposure period 6 hrs without S9 mix |
|||||
DMSO |
-- |
100 |
100 |
0 |
0.5 |
MMC |
0.1 |
78 |
97 |
0 |
52.0 |
EBS |
19.53 |
93 |
149 |
0.5 |
2.0 |
39.06 |
87 |
104 |
0 |
1.0 |
|
78.13 |
90 |
110 |
0 |
0.5 |
|
Exposure period 24 hrs without S9 mix |
|||||
DMSO |
-- |
100 |
100 |
0 |
0 |
MMC |
0.05 |
54 |
274 |
0 |
34.0 |
EBS |
9.77 |
99 |
116 |
0 |
0.5 |
19.53 |
106 |
179 |
0 |
1.5 |
|
39.06 |
110 |
168 |
0 |
1.0 |
|
Exposure period 6 hrs with S9 mix |
|||||
DMSO |
-- |
100 |
100 |
0 |
0 |
CP |
5.0 |
56 |
58 |
0 |
10.5 |
EBS |
19.53 |
87 |
97 |
0 |
0 |
39.06 |
84 |
112 |
0 |
0 |
|
78.13 |
83 |
89 |
0 |
0 |
MMC: Mitomycin C
CP: Cyclophosphamide
EBS: Ethylene Bis(stearamide)
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
In vivo:
Waiving - No in vivo testing required as none of the in vitro tests were
positive for genetic toxicity.
Link to relevant study records
- Endpoint:
- genetic toxicity in vivo
- Data waiving:
- other justification
- Justification for data waiving:
- other:
Reference
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
There is no data available on the genetic toxicity of Amides, C16 and C18-C20 (even numbered, unsaturated), N,N’-ethylenebis. In order to fulfil the standard information requirements set out in Annex VII and VIII, 8.4, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, read-across from a structurally related substance is conducted.
A detailed justification for the grouping of chemicals and read-across is provided in Sections 7.1 and 13.
In vitro
There exist three reliable studies on genetic toxicity in vitro of the structurally related substance Amides, C16-C18 (even), N,N'-ethylenebis. Therefore, read-across based on the analogue approach is performed to cover this endpoint.
- Gene mutation in bacteria
The gene mutation in bacteria was assessed by the Reverse Mutation Assay (Ames test) conducted with 5 Salmonella typhimurium strains (TA 1535, TA 1537, TA 1538, TA 98 and TA 100) and the yeast strain Saccharomyces cerevisiae D4, which was comparable to OECD guideline 471 (Jagannath, 1978). All assays were conducted with the test substance at concentrations ranging from 0.1 to 500 µg/plate in the presence and absence of a metabolic activation system (Aroclor1254-induced rat liver S9). Additionally, 1000 µg/plate was included as test concentration in a repetition test with TA 98. Although there was no strain included to detect cross-linking mutagens according to the actual version of the guideline, the study was acceptable according to the standards of the time of conduction. In this study no mutagenic effects on bacteria were observed with or without metabolic activation at any of the concentrations tested.
- Gene mutation in mammalian cells
The mutagenicity of the test substance in mammalian cells in vitro was determined in the L5178Y mouse lymphoma cell line according to OECD guideline 476 (Verspeek-Rip, 2010). The number of mutants derived from 5 exposure plates was determined in 2 independent experiments. In the first experiment, the mouse lymphoma cells were exposed for 3 h to the test substance at concentrations ranging from 0 to 1.0 µg/mL in the absence and presence of a metabolic activation system (8% (v/v) Phenobarbital/β-Naphtoflavone-induced rat liver S9-mix). Precipitation occurred at concentrations ≥ 0.6 µg/mL, no cytotoxicity was observed, and there was no significant increase of mutant frequencies at the TK locus compared to the control groups. These findings were confirmed by a second independent experiment conducted with the same test substance concentrations, but with 3-h exposure in the presence of 12% (v/v) rat liver S9-mix or 24-h exposure without metabolic activation. Therefore, it was concluded that under the conditions used in the study, the test material was not mutagenic at the TK-locus of mouse lymphoma L5178Y cells in the absence and presence of metabolic activation.
- Chromosome aberrations
The clastogenic activity of the test substance was investigated using a Chromosomal Aberration Assay in a Chinese Hamster lung (CHL) cell line according to OECD guideline 473 and in compliance with GLP (Wright, 2006). Duplicate cell cultures were exposed for 6 h to concentrations up to 156.25 µg/mL in the presence or absence of a metabolic activation system (Phenobarbital/β-Naphtoflavone-induced rat liver S9-mix), followed by an 18-h recovery period, or for 24 h to concentrations up to 78.13 µg/mL in the absence of metabolic activation. Three concentrations (up to 78.13 µg/mL) and the vehicle control were chosen for analysis of 200 metaphases. The highest concentration chosen was the lowest one at which precipitation occurred. The chromosomes were analysed for structural aberrations comprising chromosome and chromatid breaks and exchanges, gaps, numerical aberrations and multiple aberrations. Under the conditions of this study the test substance did not induce any statistically significant, dose-related increases in the frequency of cells with structural or numerical chromosome aberrations either in the presence or absence of a metabolic activation after various exposure times. The test material was therefore considered to be non-clastogenic to CHL cells in vitro.
In vivo
No study investigating the genetic toxicity in vivo of Amides, C16 and C18-C20 (even numbered, unsaturated), N,N’-ethylenebis is available. However, there exist reliable data on genetic toxicity in vitro of the structurally related substance Amides, C16-C18 (even), N,N'-ethylenebis, which are used for read-across based on the analogue approach.
According to Regulation (EC) No 1907/2006, Annex IX, column 2, testing for genetic toxicity in vivo is not indicated as the structurally closely related source substance Amides, C16-C18 (even), N,N'-ethylenebis did not demonstrate any genotoxic activity in bacteria or mammalian cells in vitro.
Based on the negative results of the available studies on the structural analogue Amides, C16-C18 (even), N,N'-ethylenebis, it may be concluded that the substance Amides, C16 and C18-C20 (even numbered, unsaturated), N,N’-ethylenebis does not induce genetic toxicity in vitro and in vivo.
Justification for classification or non-classification
The available data on genetic toxicity of a substance structurally related to Amides, C16 and C18-C20 (even numbered, unsaturated), N,N’-ethylenebis according to Regulation (EC) No 1907/2006, Annex XI, 1.5 do not meet the criteria for classification according to Regulation (EC) No 1272/2008; therefore, Amides, C16 and C18-C20 (even numbered, unsaturated), N,N’-ethylenebis is not expected to exert a genotoxic potential, either, and the data are thus conclusive but not sufficient for classification.
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