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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 620-092-1 | CAS number: 54024-17-8
- 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:
- 8 June 2020 - 10 July 2020
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 020
- Report date:
- 2020
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- May 2008
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- July 1997
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- (3aS,3bS,9aR,9bS,11aS)‐11a‐ethyl‐10‐methylidene‐1H,2H,3H,3aH,3bH,4H,5H,7H,8H,9H,9aH,9bH,10H,11H,11aH‐cyclopenta[a]phenanthrene‐1,7‐dione
- Cas Number:
- 54024-17-8
- Molecular formula:
- C20 H26 O2
- IUPAC Name:
- (3aS,3bS,9aR,9bS,11aS)‐11a‐ethyl‐10‐methylidene‐1H,2H,3H,3aH,3bH,4H,5H,7H,8H,9H,9aH,9bH,10H,11H,11aH‐cyclopenta[a]phenanthrene‐1,7‐dione
- Test material form:
- solid: particulate/powder
- Details on test material:
- Storage Conditions: In refrigerator (2-8°C)
Constituent 1
- Specific details on test material used for the study:
- Physical Description: Off-white powder
Storage conditions: In refrigerator (2-8°C)
Test item handling: No specific handling conditions required
Method
- Target gene:
- Salmonella typhimurium strain: histidine locus
Escherichia coli: tryptophan locus
Species / strainopen allclose all
- Species / strain / cell type:
- E. coli WP2 uvr A
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Metabolic activation system:
- Type and composition of metabolic activation system:
- source of S9: Rat liver microsomal enzymes (S9 homogenate) prepared from male Sprague Dawley rats that had been injected intraperitoneally with Aroclor 1254 (500 mg/kg body weight).
- method of preparation of S9 mix: S9-mix was prepared immediately before use and kept refrigerated. S9-mix contained per 10 mL: 30 mg NADP and 15.2 mg glucose-6-phosphate in 5.5 mL Milli-Q water; 2 mL 0.5 M sodium phosphate buffer pH 7.4; 1 mL 0.08 M MgCl2 solution; 1 mL 0.33 M KCl solution. The above solution was filter (0.22 μm)-sterilized.
- concentration S9 in the S9-mix: 5% (v/v) S9-fraction - Test concentrations with justification for top dose:
- Dose-range finding test: 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 μg/plate
First experiment: 52, 164, 512, 1600 and 5000 μg/plate
Second experiment:
- Without S9-mix: 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 μg/plate (TA1535, TA1537, TA100 and WP2uvrA); 1.7, 5.4, 17, 52, 164, 512 and 1600 μg/plate (TA98)
- With S9-mix: 52, 164, 512, 1600 and 5000 μg/plate (TA1535, TA1537, TA100 and WP2uvrA); 5.4, 17, 52, 164, 512 and 1600 μg/plate (TA98) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
Controlsopen allclose all
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- With metabolic activation
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 2-nitrofluorene
- sodium azide
- methylmethanesulfonate
- other: ICR-191
- Remarks:
- Without metabolic activation
- 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:
- Cell density at seeding: 10^9 cells/mL
- Test substance added in agar (plate incorporation); preincubation
TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable:
- Exposure duration: 48 ± 4 h (direct plate assay); 30 ± 2 minutes + 48 ± 4 h after solidification of the top agar (pre-incubation assay)
METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method: the reduction of the bacterial background lawn, the increase in the size of the microcolonies and the reduction of the revertant colonies were observed
METHODS FOR MEASUREMENTS OF GENOTOXICIY
- Colony counting: The revertant colonies were counted automatically with the Sorcerer Colony Counter. Plates with sufficient test item precipitate to interfere with automated colony counting were counted manually. Evidence of test item precipitate on the plates and the condition of the bacterial background lawn were evaluated when considered necessary, macroscopically and/or microscopically by using a dissecting microscope. - Evaluation criteria:
- In addition to the criteria stated below, any increase in the total number of revertants should be evaluated for its biological relevance including a comparison of the results with the historical control data range.
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 times the concurrent control, and the total number of revertants in tester strains TA1535, TA1537 or TA98 is not greater than three 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 times the concurrent control, or the total number of revertants in tester strains TA1535, TA1537 or TA98 is greater than three 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.
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- No cytotoxicity observed in experiment 1. Cytotoxicity observed in experiment 2 (without S9-mix).
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- No cytotoxicity observed in experiment 1. Cytotoxicity observed in experiment 2 (with and without S9-mix).
- Vehicle controls validity:
- valid
- 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:
- No cytotoxicity observed in experiment 1. Cytotoxicity observed in experiment 2 (with and without S9-mix).
- Vehicle controls validity:
- valid
- 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:
- No cytotoxicity observed in experiment 1. Cytotoxicity observed in experiment 2 (with and without S9-mix).
- Vehicle controls validity:
- valid
- 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:
- No cytotoxicity observed in experiment 1. Cytotoxicity observed in experiment 2 (with and without S9-mix).
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- First Experiment: Direct Plate Assay (dose-range finding study is reported as part of the first experiment):
- Precipitate: Precipitation of the test item on the plates was not observed in any tester strain.
- Toxicity: In strain TA1537 (presence of S9-mix), a fluctuation in the number of revertant colonies below the laboratory historical control data range was observed. However, since no doserelationship was observed, the reduction was not considered to be caused by toxicity of the
test item. It is more likely the reduction was caused by an incidental fluctuation in the number of revertant colonies.
- Mutagenicity: In the direct plate test, no increase in the number of revertants was observed upon treatment with the test item under all conditions tested.
Second Experiment: Pre-Incubation Assay:
- Precipitate: The test item precipitated on the plates at dose levels of 512 and/or 1600 μg/plate and above.
- Toxicity: Cytotoxicity, as evidenced by a reduction in the number of revertants, a reduction in the
bacterial lawn and/or the presence of microcolonies was observed in all tester strains in the absence and presence of S9-mix, except in tester strain WP2uvrA in the presence of S9-mix.
- Mutagenicity: In the pre-incubation test, no increase in the number of revertants was observed upon treatment with the test item under all conditions tested.
Any other information on results incl. tables
Table 1. Dose-Range Finding Test: Mutagenic Response of EMETAM in the Salmonella typhimurium Reverse Mutation Assay and in the Escherichia coli Reverse Mutation Assay
Direct Plate Assay
(µg/plate) |
| ||
|
|
|
|
Without S9-mix
Positive control | 931 | ± | 91 |
| 1504 | ± | 20 |
|
|
|
|
|
Solvent control | 97 | ± | 14 |
| 21 | ± | 3 |
|
|
|
|
|
1.7 | 100 | ± | 15 |
| 13 | ± | 3 |
|
|
|
|
|
5.4 | 94 | ± | 3 |
| 15 | ± | 2 |
|
|
|
|
|
17 | 99 | ± | 9 |
| 17 | ± | 5 |
|
|
|
|
|
52 | 103 | ± | 9 |
| 15 | ± | 4 |
|
|
|
|
|
164 | 108 | ± | 6 |
| 21 | ± | 4 |
|
|
|
|
|
512 | 91 | ± | 13 |
| 17 | ± | 4 |
|
|
|
|
|
1600 | 92 | ± | 17 |
| 14 | ± | 6 |
|
|
|
|
|
5000 | 86 | ± | 6 | n NP | 18 | ± | 6 | n NP |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
With S9-mix
Positive control | 1906 | ± | 214 |
| 195 | ± | 17 |
|
|
|
|
|
Solvent control | 98 | ± | 5 |
| 25 | ± | 4 |
|
|
|
|
|
1.7 | 81 | ± | 5 |
| 33 | ± | 12 |
|
|
|
|
|
5.4 | 83 | ± | 5 |
| 27 | ± | 3 |
|
|
|
|
|
17 | 96 | ± | 26 |
| 17 | ± | 5 |
|
|
|
|
|
52 | 96 | ± | 5 |
| 23 | ± | 4 |
|
|
|
|
|
164 | 99 | ± | 6 |
| 23 | ± | 4 |
|
|
|
|
|
512 | 97 | ± | 21 |
| 17 | ± | 4 |
|
|
|
|
|
1600 | 97 | ± | 18 |
| 20 | ± | 3 |
|
|
|
|
|
5000 | 85 | ± | 23 | n NP | 12 | ± | 4 | n NP |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
NP | No precipitate |
n | Normal bacterial background lawn |
Table 2. Experiment 1: Mutagenic Response of EMETAM in the Salmonella typhimurium Reverse Mutation Assay
Direct Plate Assay
(µg/plate) |
| ||
|
|
|
|
Without S9-mix
Positive control | 876 | ± | 56 |
| 449 | ± | 71 |
| 1252 | ± | 25 |
|
Solvent control | 9 | ± | 3 |
| 2 | ± | 1 |
| 13 | ± | 4 |
|
52 | 12 | ± | 2 |
| 3 | ± | 2 |
| 15 | ± | 4 |
|
164 | 12 | ± | 6 |
| 2 | ± | 2 |
| 16 | ± | 5 |
|
512 | 11 | ± | 4 |
| 5 | ± | 2 |
| 15 | ± | 4 |
|
1600 | 5 | ± | 1 |
| 4 | ± | 4 |
| 14 | ± | 4 |
|
5000 | 5 | ± | 2 | n NP | 2 | ± | 1 | n NP | 10 | ± | 2 | n NP |
|
|
|
|
|
|
|
|
|
|
|
|
|
With S9-mix
Positive control | 270 | ± | 12 |
| 766 | ± | 47 |
| 1291 | ± | 95 |
|
Solvent control | 13 | ± | 5 |
| 3 | ± | 2 |
| 11 | ± | 1 |
|
52 | 12 | ± | 2 |
| 4 | ± | 3 |
| 16 | ± | 9 |
|
164 | 14 | ± | 5 |
| 7 | ± | 5 |
| 17 | ± | 3 |
|
512 | 9 | ± | 5 |
| 6 | ± | 3 |
| 22 | ± | 6 |
|
1600 | 6 | ± | 2 |
| 1 | ± | 1 |
| 12 | ± | 3 |
|
5000 | 6 | ± | 1 | n NP | 2 | ± | 1 | n NP | 12 | ± | 2 | n NP |
|
|
|
|
|
|
|
|
|
|
|
|
|
NP | No precipitate |
n | Normal bacterial background lawn |
Table 3. Experiment 2: Mutagenic Response of EMETAM in the Salmonella typhimurium Reverse Mutation Assay and in the Escherichia coli Reverse Mutation Assay
Pre-incubation Assay
(µg/plate) |
| ||||
|
|
|
|
|
|
Without S9-mix
Positive control | 1045 | ± | 63 |
| 197 | ± | 27 |
| 1724 | ± | 144 |
| 628 | ± | 34 |
| 1143 | ± | 125 |
|
Solvent control | 9 | ± | 2 |
| 3 | ± | 2 |
| 13 | ± | 6 |
| 86 | ± | 19 |
| 18 | ± | 4 |
|
Positive control1 | 999 | ± | 89 |
| 165 | ± | 22 |
|
| - |
|
| 679 | ± | 48 |
|
| - |
|
|
Solvent control1 | 10 | ± | 2 |
| 4 | ± | 4 |
|
| - |
|
| 94 | ± | 9 |
|
| - |
|
|
0.171 | 11 | ± | 0 |
| 8 | ± | 4 |
|
| - |
|
| 101 | ± | 12 |
|
| - |
|
|
5.41 | 12 | ± | 3 |
| 6 | ± | 5 |
| 12 | ± | 4 |
| 112 | ± | 7 | NP |
| - |
|
|
171 | 12 | ± | 4 | n NP | 5 | ± | 2 | n NP | 17 | ± | 5 |
| 110 | ± | 15 | n NP |
| - |
|
|
52 | 9 | ± | 4 |
| 5 | ± | 3 |
| 19 | ± | 3 |
| 89 | ± | 19 | i | 14 | ± | 2 |
|
164 | 10 | ± | 5 | n | 7 | ± | 4 | n | 18 | ± | 5 | n NP | 96 | ± | 17 | n | 17 | ± | 1 |
|
512 | 4 | ± | 2 | m NP |
|
|
| e NP MC | 11 | ± | 6 | m SP |
|
| e NP MC | 12 | ± | 2 | NP | |
1600 |
|
| e SP MC |
|
| e SP MC | 12 | ± | 4 | e MP |
|
| e SP MC | 9 | ± | 2 | MP | |||
5000 |
|
| e HP |
|
| e HP |
| - |
|
|
|
| e HP |
|
| n HP |
With S9-mix
Positive control | 294 | ± | 37 |
| 382 | ± | 19 |
| 991 | ± | 71 |
| 782 | ± | 242 |
| 385 | ± | 13 |
| |||
Solvent control | 8 | ± | 4 |
| 5 | ± | 2 |
| 19 | ± | 10 |
| 96 | ± | 6 |
| 24 | ± | 7 |
| |||
5.4 | - |
|
|
| - |
|
|
| 23 | ± | 1 |
| - |
|
|
| - |
|
|
| |||
17 | - |
|
|
| - |
|
|
| 14 | ± | 2 |
| - |
|
|
| - |
|
|
| |||
52 | 6 | ± | 2 |
| 7 | ± | 4 |
| 15 | ± | 4 |
| 101 | ± | 1 |
| 20 | ± | 3 |
| |||
164 | 9 | ± | 5 |
| 5 | ± | 3 | n | 18 | ± | 6 |
| 99 |
|
| n ii | 21 | ± | 3 |
| |||
512 | 6 | ± | 5 | n NP | 5 | ± | 2 | s NP | 18 | ± | 2 | n NP | 75 | ± | 13 | m NP | 20 | ± | 4 | NP | |||
1600 | 4 | ± | 3 | s SP | 1 | ± | 2 | s SP | 14 | ± | 8 | s SP | 60 | ± | 11 | m SP | 25 | ± | 5 | SP | |||
5000 | 3 | ± | 1 | m SP | 3 | ± | 1 | s MP | - |
|
|
|
|
| m HP | 16 | ± | 3 | n MP | ||||
|
|
|
| ||||||||||||||||||||
| 1 - HP | Data from additional experiment, except for tester strain TA98 Not tested Heavy Precipitate |
| ||||||||||||||||||||
| MC | Microcolonies |
| ||||||||||||||||||||
| MP | Moderate Precipitate |
| ||||||||||||||||||||
| NP | No precipitate |
| ||||||||||||||||||||
| SP | Slight Precipitate |
| ||||||||||||||||||||
| e | Bacterial background lawn extremely reduced |
| ||||||||||||||||||||
| i | One plate infected, mean of two plates |
| ||||||||||||||||||||
| ii | Two plates infected |
| ||||||||||||||||||||
| m | Bacterial background lawn moderately reduced |
| ||||||||||||||||||||
| n | Normal bacterial background lawn |
| ||||||||||||||||||||
| s | Bacterial background lawn slightly reduced |
|
Applicant's summary and conclusion
- Conclusions:
- In an AMES test, performed according to OECD guideline 471 and in accordance with GLP principles., EMETAM is not mutagenic with or without metabolic activation.
- Executive summary:
An AMES test was performed according to OECD guideline 471 and in accordance with GLP principles. The objective of this study was to determine the potential of EMETAM and/or its metabolites to induce reverse mutations at the histidine locus in several strains of Salmonella typhimurium (S. typhimurium; TA98, TA100, TA1535, and TA1537), and at the tryptophan locus of Escherichia coli (E. coli) strain WP2uvrA in the presence or absence of an exogenous mammalian metabolic activation system (S9).
The test was performed in two independent experiments, at first a direct plate assay was performed and secondly a pre-incubation assay.In the dose-range finding study, the test item was initially tested up to concentrations of 5000 μg/plate in the strains TA100 and WP2uvrA in the direct plate assay. The test item did not precipitate on the plates at this dose level. The bacterial background lawn was not reduced at any of the concentrations tested and no biologically relevant decrease in the number of revertants was observed. Results of this dose-range finding test were reported as part of the first mutation assay.
In the first mutation experiment, the test item was tested up to concentrations of 5000 μg/plate in the strains TA1535, TA1537 and TA98. The test item did not precipitate on the plates at this dose level. The bacterial background lawn was not reduced at any of the concentrations tested and no biologically relevant decrease in the number of revertants was observed.
In the second mutation experiment, the test item was tested up to concentrations of 5000 μg/plate in the tester strains TA1535, TA1537, TA100 and WP2uvrA and up to 1600 μg/plate in the tester strain TA98 in the pre-incubation assay. The test item was tested up to or beyond a precipitating dose level. Cytotoxicity, as evidenced by a decrease in the number of revertants, reduction of the bacterial background lawn and/or the presence of microcolonies, was observed in all tester strains in the absence and presence of S9-mix, except in tester strain WP2uvrA in the presence of S9-mix.
The negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.
The test item did not induce a dose-related increase in the number of revertant (His+) colonies in each of the four tester strains (TA1535, TA1537, TA98 and TA100) and in the number of revertant (Trp+) colonies in tester strain WP2uvrA both in the absence and presence of S9-metabolic activation. These results were confirmed in a follow-up experiment.
In conclusion, based on the results of this study it is concluded that EMETAM is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.
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.