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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 948-046-9 | 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
Skin irritation / corrosion
Administrative data
- Endpoint:
- skin irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 07 October 2017 to ****
- Reliability:
- 1 (reliable without restriction)
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Reaction mass of benzenamine, 2-ethyl-N-(2-ethyl-nonylphenyl) nonyl-, branched and benzenamine, 2-ethyl-N-(2-ethyl-nonylphenyl)-, branched
- Molecular formula:
- Not applicable
- IUPAC Name:
- Reaction mass of benzenamine, 2-ethyl-N-(2-ethyl-nonylphenyl) nonyl-, branched and benzenamine, 2-ethyl-N-(2-ethyl-nonylphenyl)-, branched
- Test material form:
- liquid
- Details on test material:
- CAS number: Not yet known
EC Number: Not yet known
Constituent 1
- Specific details on test material used for the study:
- Purity: >93%
Physical state/Appearance: Dark amber viscous liquid
In vitro test system
- Test system:
- human skin model
- Source species:
- human
- Cell type:
- other: reconstructed human epidermis tissues
- Cell source:
- other: not specified
- Details on test system:
- Triplicate tissues were treated with the test item for an exposure period of 15 minutes. The test item was applied topically to the corresponding tissues ensuring uniform covering. 10 µL (26.3 µL/cm^2) of the test item was applied to the epidermis surface. Triplicate tissues treated with 10 µL of DPBS served as the negative controls and triplicate tissues treated with 10 µL of SDS 5% w/v served as the positive controls. The plates were kept in the biological safety cabinet at room temperature for 15 minutes. At the end of the exposure period, each tissue was removed from the well using forceps and rinsed using a wash bottle containing DPBS with Ca++ and Mg++. The rinsed tissues were transferred to the second column of 3 wells containing 2 mL of maintenance medium in each well. The rinsed tissues were incubated at 37 °C, 5% CO2 in air for 42 hours.
MTT Loading/Formazan Extraction (Day 3)
Following the 42 hour post-exposure incubation period each 12-well plate was placed onto a plate shaker at room temperature for 15 minutes to homogenize the released mediators in the maintenance medium. Maintenance medium from beneath each tissue was transferred to pre labeled micro tubes and stored in a freezer at 14 to 30 ºC for possible inflammatory mediator IL-1α determination.
MTT solution, freshly prepared in assay medium, was pipetted into the third column of 3 wells of the 12-well plates. The tissues were transferred to the MTT filled wells, being careful to remove any excess maintenance medium from the bottom of the tissue insert by blotting on absorbent paper. The tissues were incubated for 3 hours at 37 °C, 5% CO2 in air. At the end of the 3 hour incubation period each tissue was placed onto absorbent paper to dry. A total biopsy of the epidermis was made, the epidermis was carefully separated from the collagen matrix using forceps and both parts (epidermis and collagen matrix) placed into labeled micro tubes containing acidified isopropanol, ensuring that both the epidermis and collagen matrix were fully immersed. Each tube was plugged to prevent evaporation and mixed thoroughly on a vortex mixer and then refrigerated at 1 to 10 °C until Day 6 of the experiment, allowing the extraction of formazan crystals out of the MTT-loaded tissues. - Control samples:
- other: Dulbecco’s Phosphate Buffered Saline (DPBS) with Ca++ and Mg++
- Amount/concentration applied:
- 10 µL (26.3 µL/cm^2) of the test item was applied to the epidermis surface.
- Duration of treatment / exposure:
- Triplicate tissues were treated with the test item for an exposure period of 15 minutes.
- Duration of post-treatment incubation (if applicable):
- At the end of the exposure period each tissue was rinsed before incubating for 42 hours.
- Number of replicates:
- Three
Results and discussion
In vitro
Results
- Irritation / corrosion parameter:
- % tissue viability
- Value:
- 86.1
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- no indication of irritation
Any other information on results incl. tables
Direct MTT Reduction
The MTT solution containing the test item turned purple which indicated that the test item directly reduced MTT. Therefore, an additional procedure using water killed tissues was performed. However, the results obtained showed that negligible direct reduction of MTT occurred. It was therefore considered unnecessary to use the results of the water killed tissues for quantitative correction of results.
Assessment of Color Interference with the MTT Endpoint
The test item did not change the color of the solution. However the test item was an amber viscous liquid which may have adhered to the tissue culture surface during the main test and thus cause color interference. Therefore, as a precaution, color correction tissues were incorporated into the test to correct for this possibility. However, the results obtained showed that negligible color interference occurred (Appendix 3). It was therefore considered unnecessary to use the results of the color correction tissues for quantitative correction of results.
Test Item, Positive Control Item and Negative Control Item
The relative mean viability of the test item treated tissues was 86.1% (>50%) after a 15 Minute exposure period and 42 hour post exposure incubation period. Traces of residual test item remained on the test item treated tissue culture surfaces after rinsing due to viscosity.
Table 1: Mean Viabilities of the EPISKIN™ Tissues
Treatment |
OD570 |
Relative Tissue Viability |
Test Item |
0.701 ± 0.044 |
86.1% ± 5.4% |
DPBS (Negative Control) |
0.814± 0.049 |
100%± 6.0% |
0.5% SDS (Positive Control) |
0.302 ± 0.044 |
37.1% ± 5.4% |
Quality Criteria
The relative mean viability for tissues treated with 5% (w/v) SDS aqueous solution (positive control) was 37.1% relative to the negative control treated tissues and the standard deviation value of the viability was 5.4% (≤18%). The positive control acceptance criteria were therefore satisfied.
The mean OD570 for the negative control (DPBS) treated tissues was 0.814, within the range of 0.6 - 1.5 and the standard deviation value of the viability was 6.0% (≤18%). The negative control acceptance criteria were therefore satisfied.
The standard deviation calculated from individual tissue viabilities of the three identically test item treated tissues was 5.4% (≤18%). The test item acceptance criterion was therefore satisfied.
Applicant's summary and conclusion
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- The test item was not classified as a skin irritant according to Regulation (EC) No. 1272/2008 Classification, Labelling and Packaging of Substances and Mixtures (EU CLP).
- Executive summary:
The skin irritation potential of the test material was evaluated using the EPISKIN™ reconstructed human epidermis model. Reconstructed human epidermal cultures were treated topically in triplicate with the test material for 15 minutes, rinsed, and then incubated post-exposure for 42 hours. Cell viability was measured by means of the colorimetric MTT reduction assay, in which viability is measured by enzymatic reduction of the yellow MTT tetrazolium salt to a blue/purple formazan salt within the mitochondria of viable cells.
The test item was found to directly reduce MTT; therefore additional non-viable tissues were incorporated for correction purposes. The test item was found to have the possibility to cause color interference with the MTT endpoint; therefore, as a precaution, additional tissues were incorporated to correct for this. A third set of controls was included, comprising non-viable tissues, to prevent a double correction for a colored test item that also reduces MTT.
At the end of the 42-hour post exposure incubation period each tissue was taken for MTT-loading. After MTT-loading a total biopsy of each epidermis was made and placed into micro tubes containing acidified isopropanol for extraction of formazan crystals. At the end of the formazan extraction period each tube was mixed thoroughly, duplicate 200 μL samples were transferred to the appropriate wells of a pre-labeled 96-well plate, and the optical density was measured at 570 nm. Data are presented in the form of percentage viability (MTT reduction in the test item or positive control treated tissues relative to negative control tissues).
The relative mean viability of the test item treated tissues was 86.1%, greater than 50%, and therefore, the test item was considered as non-irritant to skin and was not classified according to GHS. The quality criteria required for acceptance of results in the test were satisfied.
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.