Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 247-161-5 | CAS number: 25646-71-3
- 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
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 18 Jan 2018 - 07 Feb 2018
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- adopted July 21, 1997
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- N-(2-(4-amino-N-ethyl-m-toluidino)ethyl)methanesulphonamide sesquisulphate
- EC Number:
- 247-161-5
- EC Name:
- N-(2-(4-amino-N-ethyl-m-toluidino)ethyl)methanesulphonamide sesquisulphate
- Cas Number:
- 25646-71-3
- Molecular formula:
- C12H21N3O2S.3/2H2O4S
- IUPAC Name:
- bis(N-{2-[(4-amino-3-methylphenyl)(ethyl)amino]ethyl}methanesulfonamide); tris(sulfuric acid)
Constituent 1
Method
- Target gene:
- his operon for S. typhimurium strains
trp operon for E. coli strains
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Species / strain / cell type:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Metabolic activation system:
- cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Arocolor 1254.
- Test concentrations with justification for top dose:
- Pre-experiment (dose-range finding test, Direct Plate Assay): 1.7, 5.4, 17, 52, 164, 512, 1600, 5000 µg/plate
Main-experiment (Pre-incubation Assay): 17, 52, 164, 512, 1600, 5000 µg/plate (Based on the results of the dose-range finding test) - Vehicle / solvent:
- - Vehicle/solvent used: water (Milli-Q-water, Millipore Corp., Bedford, MA., USA).
- Justification for choice of solvent/vehicle: The test item formed a clear colorless solution in Milli-Q water.
- Vehicle/solvent used for positive control items: Saline = physiological saline (Eurovet Animal Health, Bladel, The Netherlands)
DMSO = dimethyl sulfoxide (Merck, Darmstadt, Germany)
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 2-nitrofluorene
- sodium azide
- methylmethanesulfonate
- other: ICR-191, 2-aminoanthracene (2AA)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar, Direct Plate Assay, Pre-incubation assay
DURATION
- Preincubation period: 30 ± 2 min
- Exposure duration: at least 48 ± 4 h
NUMBER OF REPLICATIONS: 3 replications each in 2 independent experiments
DETERMINATION OF CYTOTOXICITY
- Method: To determine the toxicity of the test item, the reduction of the bacterial background lawn, the increase in the size of the microcolonies and the reduction of the revertant colonies were observed. - Evaluation criteria:
- ACCEPTABILITY CRITERIA
A Salmonella typhimurium reverse mutation assay and/or Escherichia coli reverse mutation assay is considered acceptable if it meets the following criteria:
a) The vehicle control and positive control plates from each tester strain (with or without S9-mix) must exhibit a characteristic number of revertant colonies when compared against relevant historical control data generated at Charles River Den Bosch.
b) The selected dose-range should include a clearly toxic concentration or should exhibit limited solubility as demonstrated by the preliminary toxicity range-finding test or should extend to 5 mg/plate.
c) No more than 5% of the plates are lost through contamination or some other unforeseen event. If the results are considered invalid due to contamination, the experiment will be repeated.
All results presented in the tables of the report are calculated using values as per the raw data rounding procedure and may not be exactly reproduced from the individual data presented.
EVALUATION CRITERIA
A test item is considered negative (not mutagenic) in the test if:
a) The total number of revertants in tester strain TA100 or WP2uvrA is not greater than two (2) times the concurrent control, and the total number of revertants in tester strains TA1535, TA1537 or TA98 is not greater than three (3) times the concurrent control.
b) The negative response should be reproducible in at least one follow up experiment. A test item is considered positive (mutagenic) in the test if:
a) The total number of revertants in tester strain TA100 or WP2uvrA is greater than two (2) times the concurrent control, or the total number of revertants in tester strains TA1535, TA1537 or TA98 is greater than three (3) times the concurrent control.
b) In case a repeat experiment is performed when a positive response is observed in one of the tester strains, the positive response should be reproducible in at least one follow up experiment. - Statistics:
- Mean values and standard deviations were calculated.
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Cytotoxicity was observed at 1600 μg/plate onwards without metabolic activation and at 5000 μg/plate with metabolic activation.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Remarks:
- 2.3-fold dose-related increase in the number of revertant colonies in the absence of S9-mix.
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Cytotoxicity was observed at 1600 μg/plate onwards without metabolic activation and at 5000 μg/plate with metabolic activation.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Remarks:
- 1.9-fold dose-related increase in the number of revertant colonies in the presence of S9-mix. The increase observed was within the laboratory historical control data ranges and less than two-fold the concurrent solvent control
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Cytotoxicity was observed at 1600 μg/plate onwards without metabolic activation and at 5000 μg/plate with metabolic activation.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Cytotoxicity was observed at 1600 μg/plate onwards without metabolic activation and at 5000 μg/plate with metabolic activation.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- 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:
- Cytotoxicity was observed at 1600 μg/plate onwards without metabolic activation and at 5000 μg/plate with metabolic activation.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- Direct Plate Assay
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation of the test substance on the plates was not observed at the start or at the end of the incubation period in any tester strain.
ADDITIONAL INFORMATION ON CYTOTOXICITY:
- Measurement of cytotoxicity used: Cytotoxicity, as evidenced by a decrease in the number of revertants and/or reduction of the bacterial background lawn, was observed in all tester strains. Except for tester strain WP2uvrA, where no toxicity was observed in the absence or presence of S9-mix.
Pre-Incubation Assay
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation of the test substance on the plates was not observed at the start or at the end of the incubation period in any tester strain.
ADDITIONAL INFORMATION ON CYTOTOXICITY:
- Measurement of cytotoxicity used: Cytotoxicity, as evidenced by a decrease in the number of revertants and/or reduction of the bacterial background lawn, was observed in all tester strains from 1600 μg/plate onwards without metabolic activation and at 5000 μg/plate with metabolic activation. Except for tester strain WP2uvrA in the presence of S9-mix, where no toxicity was observed
Any other information on results incl. tables
Table 3: Dose-Range Finding Test (Direct-Plate assay)
|
EXPERIMENT 1 (Revertant colonies per plate ± SD) |
||
|
S9-Mix |
Without
|
|
|
Test item (µg/plate) |
TA 100
|
WP2uvrA
|
Vehicle Control |
|
113 ± 11 |
33 ± 11 |
Test Substance |
1.7 |
121 ± 15 |
25 ± 5 |
5.4 |
123 ± 19 |
37 ± 2 |
|
17 |
115 ± 12 |
21 ± 1 |
|
52 |
134 ± 12 |
33 ± 6 |
|
164 |
141 ± 13 |
37 ± 6 |
|
512 |
166 ± 16 |
35 ± 0 |
|
1600 |
292 ± 22n |
41 ± 9 |
|
5000 |
0 ± 0aNP |
51 ± 4nNP |
|
Positive Control |
|
744 ± 65 |
1602 ± 77 |
|
S9-Mix |
With
|
|
|
Test item (µg/plate) |
TA 100 |
WP2uvrA |
Vehicle Control |
- |
118 ± 6 |
43 ± 7 |
Test Substance |
1.7 |
100 ± 8 |
50 ± 6 |
5.4 |
106 ± 13 |
40 ± 3 |
|
17 |
116 ± 13 |
35 ± 9 |
|
52 |
122 ± 6 |
38 ± 8 |
|
164 |
135 ± 18 |
43 ± 4 |
|
512 |
169 ± 21 |
58 ± 5 |
|
1600 |
310 ± 19 |
75 ± 4 |
|
5000 |
106 ± 941NP |
52 ± 5nNP |
|
Positive Control |
|
2156 ± 227 |
558 ± 54 |
|
1=Revertants were only observed on a small part of the plate with normal background, no bacterial growth or background was observed on a large part of the plate NP= No precipitate a = Bacterial background lawn absent n = Normal bacterial background lawn s = Bacterial background lawn slightly reduced |
Table 4: Direct-Plate assay
|
EXPERIMENT 1 (Revertant colonies per plate ± SD) |
|||
|
S9-Mix |
Without
|
||
|
Test item (µg/plate) |
TA 1535 |
TA 1537 |
TA 98 |
Vehicle Control |
|
12 ± 2 |
3 ± 2 |
11 ± 1 |
Test Substance |
17 |
12 ± 2 |
3 ± 0 |
17 ± 5 |
52 |
13 ± 5 |
4 ± 3 |
14 ± 3 |
|
164 |
14 ± 4 |
2 ± 1 |
11 ± 3 |
|
512 |
13 ± 2 |
2 ± 1 |
10 ± 6 |
|
1600 |
11 ± 3n |
2 ± 1n |
12 ± 6n |
|
5000 |
0 ± 0aNP |
2 ± 2eNP |
0 ± 0aNP |
|
Positive Control |
|
1091 ± 65 |
834 ± 49 |
1141± 30 |
|
S9-Mix |
With
|
||
|
Test item (µg/plate) |
TA 1535 |
TA 1537 |
TA 98 |
Vehicle Control |
- |
11 ± 6 |
3 ± 4 |
16 ± 2 |
Test Substance |
17 |
12 ± 5 |
4 ± 3 |
16 ± 3 |
52 |
10 ± 2 |
7 ± 4 |
22 ± 6 |
|
164 |
13 ± 5 |
4 ± 1 |
16 ± 4 |
|
512 |
17 ± 8 |
3 ± 2 |
17 ± 4 |
|
1600 |
18 ± 3 |
3 ± 2 |
19 ± 7 |
|
5000 |
5 ± 81NP |
1 ± 21NP |
3 ± 21NP |
|
Positive Control |
|
408 ± 32 |
374± 108 |
1298± 195 |
|
1=Revertants were only observed on a small part of the plate with normal background, no bacterial growth or background was observed on a large part of the plate NP= No precipitate a = Bacterial background lawn absent n = Normal bacterial background lawn s = Bacterial background lawn slightly reduced |
Table 5: Plate-Incubation Test
|
EXPERIMENT 2 (Revertant colonies per plate ± SD) |
|||||
|
S9-Mix |
Without
|
||||
|
Test item (µg/plate) |
TA 1535 |
TA 1537 |
TA 98 |
TA 100 (fold) |
WP2uvrA (fold) |
Vehicle Control |
|
10 ± 2 |
3 ± 4 |
18 ± 3 |
106 ± 14 |
19 ± 5 |
Test Substance |
17 |
13 ± 5 |
3 ± 1 |
17 ± 6 |
122 ± 15 |
28 ± 2 |
52 |
12 ± 2 |
3 ± 2 |
13 ± 5 |
130 ± 5 |
29 ± 15 (1.5) |
|
164 |
19 ± 1 |
3 ± 2 |
14 ± 3 |
134 ± 16 (1.3) |
35 ± 7 (1.8) |
|
512 |
13± 3n |
8 ± 2n |
17 ± 3n |
195 ± 5n (1.8) |
44 ± 8n(2.3) |
|
1600 |
0 ± 0a |
0± 0a |
0± 0a |
0± 0a |
1± 01n |
|
5000 |
0 ± 0aNP |
0 ± 0aNP |
0 ± 0aNP |
0 ± 0aNP |
0 ± 0aNP |
|
Positive Control |
|
1036 ± 33 |
138 ± 14 |
1146 ± 68 |
781 ± 11 |
152 ± 85 |
|
S9-Mix |
With
|
||||
|
Test item (µg/plate) |
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
WP2uvrA |
Vehicle Control |
- |
11 ± 2 |
5 ± 3 |
19 ± 6 |
102 ± 2 |
30 ± 7 |
Test Substance |
17 |
13 ± 2 |
4 ± 2 |
15 ± 6 |
166 ± 18 |
36 ± 6 |
52 |
10 ± 5 |
7 ± 2 |
13 ± 2 |
180 ± 20 |
42 ± 6 |
|
164 |
14 ± 3 |
7 ± 2 |
19 ± 4 |
139 ± 7 |
39 ± 8 |
|
512 |
10 ± 0 |
5 ± 3n |
13 ± 2 |
190 ± 26 (1.9) |
42 ± 4 (1.4) |
|
1600 |
14 ± 6 |
2 ± 1s |
15 ± 3n |
304 ± 22n(3.0) |
56 ± 13 (1.9) |
|
5000 |
0 ± 11nNP |
0 ± 0aNP |
0 ± 0aNP |
0 ± 0aNP |
57 ± 11nNP(1.9) |
|
Positive Control |
|
247 ± 22 |
230 ± 32 |
762 ± 14 |
1586 ± 89 |
587 ± 30 |
|
1=Revertants were only observed on a small part of the plate with normal background, no bacterial growth or background was observed on a large part of the plate NP= No precipitate a = Bacterial background lawn absent n = Normal bacterial background lawn s = Bacterial background lawn slightly reduced |
Applicant's summary and conclusion
- Conclusions:
- Under the conditions of the conducted test the substance induced significant dose-related increases in the number of revertant colonies in the tester strain TA100 and WP2uvrA both in the absence and presence of S9-mix. In the latter, only in the absence of S-9 mix in experiment 2, these increases exceeded the two-fold threshold (2.3-fold). Based on these results, the test substance is regarded to be mutagenic in bacteria.
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.