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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 943-330-9 | CAS number: -
- 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 mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 11 Jan - 08 Feb 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 017
- Report date:
- 2017
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 490 (In Vitro Mammalian Cell Gene Mutation Tests Using the Thymidine Kinase Gene)
- Version / remarks:
- 29 July 2016
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- other: mammalian cell gene mutation assay
Test material
- Reference substance name:
- Reaction mass of bis[2-[2-(2-butoxyethoxy)ethoxy]ethyl]adipate and 2-[2-(2-butoxyethoxy)ethoxy]ethyl(3,6,9,12-tetraoxahexadecyl)adipate
- Molecular formula:
- C26H50O10 C28H54O11
- IUPAC Name:
- Reaction mass of bis[2-[2-(2-butoxyethoxy)ethoxy]ethyl]adipate and 2-[2-(2-butoxyethoxy)ethoxy]ethyl(3,6,9,12-tetraoxahexadecyl)adipate
Constituent 1
Method
- Target gene:
- TK locus
Species / strain
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- CELLS USED
- Source of cells: JHSF (Japan Health Science Foundation)
MEDIA USED
- Type and identity of media: RPMI 1640 medium supplemented with horse serum, sodium pyruvate and penicillin/streptomycin
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically 'cleansed' against high spontaneous background: yes
- 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 for 5 days
- Test concentrations with justification for top dose:
- Cytotoxicity test, short-term treatment (4 h), with and without metabolic activation: 5000, 2500, 1250, 625, 312, 156, 78 and 39 µg/mL
Cytotoxicity test, long-term treatment (24 h), without metabolic activation: 1000, 500, 250, 125, 62.5, 31.25, 15.63 and 7.8 µg/mL
In each assay 4 doses were selected for the gene mutation test:
Short-term treatment (4 h), without metabolic activation: 625, 312, 156 and 78 µg/m:
Short-term treatment (4 h), with metabolic activation: 2500, 1250, 625 and 312 µg/mL
Long-term treatment (24 h), without metabolic activation: 500, 250, 125 and 62.5 µg/mL
Considering that the test item is a multi-constituent substance, the maximum concentration tested was 5000 µg/mL. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: culture medium
Controls
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- culture medium
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- methylmethanesulfonate
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION:
Short-term treatment (4 h), with and without metabolic activation: 5000, 2500, 1250, 625, 312, 156, 78 and 39 µg/mL
Long-term treatment (24 h), without metabolic activation: 1000, 500, 250, 125, 62.5, 31.25, 15.63 and 7.8 µg/mL
EXPERIMENTAL PROCEDURE AND DETAILS:
Short-term treatment (4 h), -/+S9:
- A series of tubes containing 6x10E+6 cells in suspension/tube in medium was used. The different concentrations of the test item were added directly into the tubes (2 cultures per concentration).
- At the end of the incubation period of 4 h, the tubes were centrifuged and the medium was aspirated. 2 washes were performed and 10 ml of medium were deposited in each culture. A count of the cells was performed on each tube, then treated cells were transferred into flasks and incubated 24 h (± 1 h) at 37 °C (± 0.5 °C), 5% CO2.
- After the incubation period, cell counting was performed and cell cultures were adjusted if necessary to 2x10E+5 cells/mL and incubated for 24 h (± 1 h) at 37 °C (± 0.5 °C), 5% CO2.
- After the incubation period, cell counting was performed again, i.e. 48 h after the end of the treatment. These counts allowed assessing the cytotoxicity expressed as a percentage of relative cell growth (RSG). The actual gene mutation test was performed using 4 selected concentrations of the test item as determined by cytotoxicity.
- The cells were seeded in 96-well plates at 2 cells/200µL in non-selective culture medium (2 plates/concentration) to determine viability. Plates were incubated at 37 °C (± 0.5 °C), 5% CO2 for 10 to 12 days. Empty wells were recorded to calculate the survival rate (cloning efficiency, CE) to determine the viability.
- In parallel, 96-well plates were seeded at 2x10E+3 cells/200µL in selective culture medium to determine the mutation frequency (4 plates/concentration). Plates were incubated at 37 °C (± 0.5 °C), 5% CO2 for 10 to 12 days. Empty wells were recorded to calculate the mutation frequency.
Long-term treatment (24 h), -S9:
- A series of flasks containing 1.5x10E+6 cells/flask in culture medium was used. The different concentrations of the test item were added directly to the flasks (2 cultures per concentration) and then incubated for 24 h (± 1 h) at 37 °C (± 0.5 °C), 5% CO2.
- After the incubation period, the cultures were transferred into sterile tubes, centrifuged and the medium was aspirated. 2 washes were performed and 10 ml of medium were deposited in each culture. A count of the cells was performed on each tube. The treated cells were placed in new culture flasks at 2x10E+5 cells/mL and incubated for 24 h (± 1 h) at 37 °C (± 0.5 °C), 5% CO2.
- All further steps are identical to the procedure for the short-term treatment (4 h).
SELECTION AGENT (mutation assays): trifluorothymidine (TFT)
NUMBER OF REPLICATIONS: duplicates
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth (cytotoxicity corresponds to relative survival compared to the respective negative control values)
OTHER EXAMINATIONS:
Cloning efficiency was determined by seeding exposed cells in one microtiter plate with a density of 2 cells/well in medium without TFT.
Small and large colonies were differentiated as small colonies are capable to indicate chromosomal mutations. - Evaluation criteria:
- A test item is considered potentially mutagenic if all of the following conditions are met:
- The mutation rate induced is significantly higher than that of the negative control,
- The observed effect is dose-dependent,
- The observed effect is reproducible,
- The rate of mutation is greater than the value of 126 plus the mutation frequency of the negative control (Moore et al., 2006).
The evaluation results are based on statistical methods, but the statistical significance should not be the only deciding factor.
If the criteria given above are not met, the test item is considered not mutagenic in this test.
Moore et al. (2006). Env. Mol. Mut. 2006, 47: 1-5 - Statistics:
- The calculations and summary of the results were performed using the Excel software. The mutant frequency for each series studied was subjected to a Chi-2 test.
Results and discussion
Test results
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- -S9: starting at 1250 µg/mL (4 h) and 1000 µg/mL (24 h); +S9: starting at 5000 µg/mL (4 h)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- VALIDATION AND COMPARISON WITH HISTORICAL CONTROL DATA:
The negative control shows a cloning efficiency (CE) between 65 and 120%, a mutation frequency (MF) between 5x10E-5 and 17x10E-5) cells and a suspension growth (SG) between 8 and 32 (4 h-treatment) and between 32 and 180 (24 h-treatment).
The positive control shows a mutation frequency (MF) higher than 3x10E-4 with at least 40% in small colonies.
Except for the positive control for the long-term treatment (24 h), without S9, these values are slightly outside the IC (inhibition concentration) 95% of the historical control data. However, they are all consistent with the validity criteria and no deviation to the study plan or incidents have been observed during the study. The control values will, therefore, be added to the historical control data of the laboratory. The results validate the test.
Any other information on results incl. tables
Tables and further details on results are included in a separeate pdf document attached to this robust study summary.
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results: negative
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.