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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 614-657-1 | CAS number: 68609-08-5
- 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:
- from 2007-10-16 to 2007-12-11
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 007
- Report date:
- 2007
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- 2000/32/EEC
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Version / remarks:
- June 1996/August 1998
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Reaction products of 3-aminomethyl-3,5,5-trimethylcyclohexylamine with 2,2'-[(1-methylethylidene)bis(4,1-phenyleneoxymethylene)]bisoxirane
- EC Number:
- 614-657-1
- Cas Number:
- 68609-08-5
- Molecular formula:
- C41H68N4O4
- IUPAC Name:
- Reaction products of 3-aminomethyl-3,5,5-trimethylcyclohexylamine with 2,2'-[(1-methylethylidene)bis(4,1-phenyleneoxymethylene)]bisoxirane
Constituent 1
Method
- Target gene:
- The Salmonella typhimurium histidine (his) reversion system measures his- -> his+ reversions. The Salmonella typhimurium strains are constructed to differentiate between base pair (TA 1535, TA 100) and frameshift (TA 1537, TA 98) mutations. The Escherichia coli WP2 uvrA (trp) reversion system measures trp– -> trp+ reversions. The Escherichia coli WP2 uvrA detect mutagens that cause other base-pair substitutions (AT to GC).
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- E. coli WP2 uvr A
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 Mix
- Test concentrations with justification for top dose:
- In the above experimental phases the examined concentration levels were different. The test item was dissolved in Dimethyl sulfoxide (DMSO).
In the Plate Incorporation Tests the investigated concentration ranges (summarised): in case of Salmonella typhimurium strains 100-0.003 μg/plate with and without metabolic activation (± S9 Mix); in Escherichia coli WP2 uvrA: 3419-0.158 μg/plate (± S9 Mix).
In the Pre-Incubation Tests: in the Salmonella typhimurium strains 10-0.003 μg/plate (–S9 Mix) and 100-0.003 μg/plate (+S9 Mix); in case of Escherichia coli WP2 uvrA 158.1-0.158 μg/plate (± S9 Mix). - Vehicle / solvent:
- Dimethyl sulfoxide DMSO
Controlsopen allclose all
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: 4-Nitro-1,2-phenylenediamine, NPD
- Remarks:
- TA 98 without S9
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- TA 100, TA 1535 without S9 Migrated to IUCLID6: NaN3
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- TA 1537 without S9 Migrated to IUCLID6: 9AA
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Remarks:
- E. coli WP2uvrA without S9 Migrated to IUCLID6: MMS
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene, 2AA
- Remarks:
- S. typhimurium TA 100, TA 98, TA 1535, TA 1537; E. coli WP2uvrA with S9
- Evaluation criteria:
- The colony numbers for control, positive control and the test plates were determined, the mean values and appropriate standard deviations were calculated.
The mutation factor (MF) was calculated by dividing the mean value of the revertant counts by the mean values of the solvent control (exact, not rounded values were used for this calculation). - Statistics:
- not applicable
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- >= 3.162 µg/plate without S9; <= 31.62 µg/plate with S9
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- >= 3.162 µg/plate without S9; <= 31.62 µg/plate with S9
- Vehicle controls validity:
- valid
- Untreated negative 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:
- >= 3.162 µg/plate without S9; <= 31.62 µg/plate with S9
- Vehicle controls validity:
- valid
- Untreated negative 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:
- >= 3.162 µg/plate without S9; <= 31.62 µg/plate with S9
- Vehicle controls validity:
- valid
- Untreated negative 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:
- >= 3.162 µg/plate without S9; <= 31.62 µg/plate with S9
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Five bacterial strains, Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and Escherichia coli WP2 uvrA were used to investigate the mutagenic potential of the test item in plate incorporation tests (Initial Mutation Test, Complementary Plate Incorporation Test, Second Complementary Plate Incorporation Test) and in pre-incubation tests (Confirmatory Mutation Test and Complementary Pre-Incubation Test).
In general, the pre-incubation method is more sensitive than the plate incorporation method. Each assay was conducted with and without metabolic activation (S9 Mix). The concentrations, including the controls, were tested in triplicate (positive and negative controls were run concurrently). In the Complementary Pre-Incubation Test only the activated part (+S9 Mix) of assay was examined. The Complementary Pre-Incubation Test was performed with Salmonella typhimurium strains.
Following concentrations of Reaction Product of Bisphenol A (BADGE) with IPDA were tested in different experiments: in the Plate Incorporation Tests the investigated concentration ranges (summarised): in case of Salmonella typhimurium strains 100-0.003 μg/plate without and with addition of metabolic activation (±S9 Mix), in Escherichia coli WP2 uvrA: 3419-0.158 μg/plate (± S9 Mix).
In the Pre-Incubation Tests: in the Salmonella typhimurium strains 10-0.003 μg/plate (–S9 Mix) and 100-0.003 μg/plate (+S9 Mix); in case of Escherichia coli WP2 uvrA 158.1-0.158 μg/plate (± S9 Mix).
In the Initial Mutation Test, in case of Salmonella typhimurium strains inhibition was observed in the examined concentration range (1-0.003 μg/plate) neither in presence, nor in absence of metabolic activation (S9 Mix).
The observed revertant colony numbers were slightly lower than the revertant colony numbers of the solvent control plates at different concentrations in case of Salmonella typhimurium TA 98, TA 1535 and TA 1537; however these variations were of the minor intensity and were considered to reflect the biological variability of the test.
In Escherichia coli WP2 uvrA the examined concentration range was higher (3419–1 μg/plate) than in the Salmonella typhimurium strains. In case of Escherichia coli WP2 uvrA no bacterial growth was observed in the concentration range of 3419-100 μg/plate without and in the range of 3419-341.9 μg/plate with addition of metabolic activation. The revertant colony numbers were reduced compared to the revertant colony numbers of the solvent control plates and the background lawn development was inhibited at the concentration of 34.19 μg/plate without, and at 100 μg/plate with addition of metabolic activation. The slightly lower revertant counts (compared to the solvent control plates) at the concentrations of 10 and 3.419 μg/plate (–S9 Mix), furthermore at 34.19 μg/plate (+S9 Mix) were evaluated as reflecting the biological variability in the test.
In the Complementary Plate Incorporation Test the examined concentration levels were 10, 3.162 and 1 μg/plate in the Salmonella typhimurium strains and 50; 15.81; 5; 1.581; 0.500 and 0.158 μg/plate in case of Escherichia coli WP2 uvrA. Unequivocally inhibitory effect (indicated by reduced revertant colony numbers compared to the solvent control plates and reduced background lawn development) of the test item was observed in all Salmonella typhimurium strains at the concentration of 10 μg/plate, and in the Escherichia coli WP2 uvrA at 50 μg/plate, without metabolic activation. Similarly to the Initial Mutation Test the slightly lower than the revertant colony numbers of the solvent control plates at different concentration levels in the examined strains were considered to reflect the biological variability of the test.
Since the cytotoxicity of the test item altered in presence of metabolic activation system (S9 Mix), a Second Complementary Plate Incorporation Test was carried out using the concentration levels of 100 and 31.62 μg/plate in Salmonella typhimurium strains and 500 and 158.1 μg/plate in Escherichia coli WP2 uvrA.
No bacterial growth was observed in the bacterial strains at the above concentration levels in absence of metabolic activation furthermore at 100 μg/plate (Salmonella typhimurium strains) and 500 and 158.1 μg/plate (Escherichia coli WP2 uvrA) in presence of metabolic activation. Furthermore reduced revertant colony number and background lawn development was observed in Salmonella typhimurium TA 98 and TA 1537 at the concentration of 31.62 μg/plate in presence of metabolic activation.
In the Confirmatory Mutation Test (Pre-Incubation Test) the examined concentration levels were: 10; 3.162; 1; 0.316; 0.1; 0.032; 0.01 and 0.003 μg/plate in the Salmonella typhimurium strains and 158.1; 50; 15.81; 5; 1.581; 0.500 and 0.158 μg/plate in case of Escherichia coli WP2 uvrA.
Strong inhibitory effects of the test item were observed in all examined strains, however in Salmonella typhimurium strains only the non-activated part (–S9 Mix) was inhibited. The signs of the inhibitory effects included reduced revertant colony numbers, compared to the solvent control plates, reduced background lawn development and pinpoint colony appearance. No bacterial growth was observed in the Salmonella typhimurium TA 98, TA 1535, TA 1537 strains at 10 μg/plate (–S9 Mix), in Escherichia coli WP2 uvrA at 158.1 and 50 μg/plate (–S9 Mix). In Escherichia coli WP2 uvrA at 158.1 μg/plate (+S9 Mix) only one revertant colony was counted on the three parallels.
In the case of Salmonella typhimurium TA 1537 at 3.162 g/plate (–S9 Mix) the revertant colony numbers were increased, but reduced background lawn development was observed and the colonies were small, pinpoint colonies. The revertant colony count were lower than the revertant count of the solvent control plates in several cases, however they were in the historical control data range in all cases.
A Complementary Pre-Incubation Test was performed to reach the cytotoxic level in presence of metabolic activation (+S9 Mix) in case of Salmonella typhimurium strains. The examined concentration levels were 100 and 31.62 μg/plate. No bacterial growth was observed in Salmonella typhimurium TA 98, TA 1535, and TA 1537 at the concentration level of 100 μg/plate, and inhibition was observed (reduced revertant colony numbers, compared to the solvent control plates, reduced background lawn development and pinpoint colony appearance) in all strains at 31.62 μg/plate, furthermore in Salmonella typhimurium TA 100 at 100 μg/plate.
In the consecutive mutation tests the inhibitory, cytotoxic effect of the test item independently of the examined strains (Salmonella typhimurium or Escherichia coli) appeared down to and including the concentration level of 3.162 μg/plate in absence and down to and including the concentration level of 31.62 μg/plate in presence of metabolic activation.
Positive and negative controls were run concurrently. The revertant colony numbers of solvent control plates without S9 Mix were within the historical control data range. The reference mutagens showed a distinct increase of induced revertant colonies. - Remarks on result:
- other: all strains/cell types tested
Applicant's summary and conclusion
- Conclusions:
- The reported data of this mutagenicity assay shows, that under the experimental conditions reported, the test item did not induce gene mutations by frameshift or base-pair substitution in the genome of the strains used. Therefore, the test item is considered non-mutagenic in this bacterial reverse mutation assay.
- Executive summary:
Experimental Phases of the Study
The test item was examined in the following phases: in Plate Incorporation Tests (Initial Mutation Test, Complementary Plate Incorporation Test, and Second Complementary Plate Incorporation Test) and Pre-Incubation Tests (Confirmatory Mutation Test and Complementary Pre-Incubation Test).
Exposure Concentrations
In the above experimental phases the examined concentration levels were different. The test item was dissolved in Dimethyl sulfoxide (DMSO).
In the Plate Incorporation Tests the investigated concentration ranges (summarised): in case of Salmonella typhimurium strains 100 - 0.003 μg/plate without and with addition of metabolic activation (± S9 Mix), in Escherichia coli WP2 uvrA: 3419 - 0.158 μg/plate (± S9 Mix). In the Pre-Incubation Tests: in the Salmonella typhimurium strains 10 - 0.003 μg/plate (–S9 Mix) and 100 - 0.003 μg/plate (+S9 Mix); in case of Escherichia coli WP2 uvrA 158.1 - 0.158 μg/plate (± S9 Mix).
Bacteria
Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 strains and Escherichia coli WP2 uvrA were used.
Results and Discussion:
Five bacterial strains, Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and Escherichia coli WP2 uvrA were used to investigate the mutagenic potential of the test item in independent experiments, in plate incorporation tests and in a pre-incubation tests. Each assay (except the Complementary Pre-Incubation Test) was conducted with and without metabolic activation (S9 Mix). The concentrations, including the controls, were tested in triplicate.
Inhibitory, cytotoxic effects of the test item were observed in the experiments in all examined bacterial strains and included reductions of spontaneous mutation rates, when compared to the solvent control values, reduced background lawn development and appearances of small pinpoint colonies. Furthermore no bacterial growth was observed for the strains at several concentration levels, with and without metabolic activation. Inhibition was stronger in the non-activated part (–S9 Mix) of the experiments.
No significant increases in revertant colony numbers of any of the five test strains were observed following treatment with the test item at any concentration level, either in the presence or absence of metabolic activation (S9 mix) in the performed experiments. Further, there was no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance in the performed experiments.
Positive and negative controls were run concurrently. The revertant colony numbers of solvent control plates without S9 Mix were within the historical control data range. The reference mutagens showed a distinct increase of induced revertant colonies.
Conclusion:
The reported data of this mutagenicity assay shows, that under the experimental conditions reported, the test item did not induce gene mutations by frameshift or base-pair substitution in the genome of the strains used. Therefore, the test item is considered non-mutagenic in this bacterial 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.