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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 824-772-0 | CAS number: 2060540-82-9
- 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 sensitisation
Administrative data
- Endpoint:
- skin sensitisation: in vitro
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 10 February 2017 - 10 March 2017
- 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 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method)
- Version / remarks:
- February 2015
- Deviations:
- yes
- Remarks:
- Except that no solubility test was performed since the test item was already formulated in water. This deviation to the guideline was considered not to have any impact on the integrity of the study once the test item was soluble in a recommended vehicle.
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- activation of keratinocytes
Test material
Reference
- Name:
- Unnamed
- Type:
- Constituent
- Test material form:
- liquid
In vitro test system
- Details on the study design:
- Vehicle: water for injections
Negative control: DMSO; this negative control was applied to cells at a concentration of 1% in culture medium.
Positive control item: Cinnamic Aldehyde (CA). For each run, the positive control item was dissolved in DMSO to a final concentration of 200 mM. This solution was then further diluted to a final concentration of 6.4 mM. It was diluted in DMSO by serial dilutions in the Master plate 100x, using a dilution factor of two, to obtain a total of five concentrations. Subsequently, each formulation of the Master plate 100x was diluted 25-fold in treatment medium in another 96-well plate called "Master plate 4x". The final tested concentrations ranged from 4 to 64 µM. All these formulations were prepared within 4 hours before use, then kept at room temperature and protected from light until use.
Criteria: interpretation of results
Acceptance criteria
Each run was considered valid if the following criteria were met:
- the positive control results should be positive, thus the gene induction should be statistically significant above the threshold of 1.5 in at least one of the tested concentrations,
- the average EC1.5 value for the positive control should be within two standard deviations of the historical mean. In addition, the average induction (Imax) in the three replicate plates for the positive control at 64 µM should be between two and eight. If the latter criterion was not fulfilled, the dose response of Cinnamic Aldehyde was carefully checked, and the run was accepted if there was a clear dose-response with increasing luciferase activity at increasing concentrations for the positive control,
- the average Coefficient of Variation of the luminescence reading in the negative control wells of the triplicate plates should be < 20%.
Results and discussion
In vitro / in chemico
Resultsopen allclose all
- Run / experiment:
- other: mean (first run)
- Parameter:
- other: EC1.5
- Remarks:
- expressed in µM
- Value:
- 4.27
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- positive indication of skin sensitisation
- Run / experiment:
- other: mean (second run)
- Parameter:
- other: EC1.5
- Remarks:
- expressed in µM
- Value:
- 13.72
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- positive indication of skin sensitisation
- Run / experiment:
- other: first run
- Parameter:
- other: Imax
- Value:
- 2.16
- Run / experiment:
- other: Second run
- Parameter:
- other: Imax
- Value:
- 3.97
- Other effects / acceptance of results:
- First and second runs
All acceptance criteria were fulfilled for the positive and negative controls. The runs were therefore considered to be valid.
The results met the criteria of a positive response in both runs.
Applicant's summary and conclusion
- Interpretation of results:
- study cannot be used for classification
- Remarks:
- The two major constituents of the test item (dodecylbenzenesulfonic acid/CAS 27176-87-0 and cyclohexanamine/CAS 108-91-8) are classified H314 “causes severe skin burns and eye damage” according to the classification provided by companies to ECHA in REACH registrations
- Conclusions:
- Under the experimental conditions of this study, the test item was positive in the KeratinoSens assay and therefore was considered to activate the Nrf2 transcription factor, though with caution due to the H314 classification ("causes severe skin burns and eye damage") of the two major constituents of the test item.
- Executive summary:
The objective of this study was to evaluate the potential of the test item to activate the Nrf2 transcription factor. This test is a part of a tiered strategy for the evaluation of skin sensitisation potential. Thus, data generated with the present Test Guideline should be used to support the discrimination between skin sensitizers and non-sensitizers in the context of an integrated approach to testing and assessment.
The study was performed according to the international OECD guideline No.442D (except that no solubility test was performed since the test item was already formulated in water) and in compliance with the principles of Good Laboratory Practice.
Principle
This in vitro test uses the KeratinoSens cell line, an immortalized and genetically modified Human adherent HaCaT keratinocyte cell line. The KeratinoSens cell line is stably transfected with a plasmid containing a luciferase gene under the transcriptional control of the SV40 origin of replication promoter. This promoter is fused with an ARE sequence. Sensitizers with electrophilic properties provoke the dissociation of Keap-1 from the transcription factor Nrf2. The free Nrf2 binds to the ARE sequence contained in the plasmid and therefore induces transcription of firefly luciferase.
Methods
The KeratinoSens cells were first plated on 96-well plates and grown for 24 hours at 37°C. Then the medium was removed and the cells were exposed to the vehicle control or to different concentrations of test item and of positive controls. The treated plates were then incubated for 48 hours at 37°C. At the end of the treatment, cells were washed and the luciferase production was measured by flash luminescence. In parallel, the cytotoxicity was measured by a MTT reduction test and was taken into consideration in the interpretation of the sensitisation results. Two independent runs were performed.
Results
First run
All acceptance criteria were met for the positive and negative controls, this run was therefore considered as validated.
This run was performed using the following concentrations 0.98, 1.95, 3.91, 7.81, 15.63, 31.25, 62.5, 125, 250, 500, 1000 and 2000 µM in culture medium containing 1% DMSO and 1% water.
At these tested concentrations:
. no precipitate/emulsion was observed in any wells at the end of the 48-hour treatment period at any tested concentrations,
. a decrease in cell viability (i.e. cell viability < 70%) was noted at concentrations = 15.63 µM,
. the corresponding IC30 and IC50 were calculated to be 11.58 and 12.73 µM, respectively,
. a statistically significant gene-fold induction above the threshold of 1.5 was noted in comparison to the negative control at 7.81 µM with an apparent overall dose-response relationship up to the cytotoxic concentrations,
. the Imax was 2.16 and the calculated EC1.5 was 4.27 µM.
Thus, the results met the criteria of a positive response
Second run
All acceptance criteria were met for the positive and negative controls, this run was therefore considered to be valid.
This run was performed using the following concentrations 1.43, 2.01, 2.84, 4.00, 5.64, 7.95, 11.2, 15.8, 22.3, 31.4, 44.3 and 62.5 µM in culture medium containing 1% DMSO and 1% water.
At these tested concentrations:
. no precipitate/emulsion was observed in any wells at the end of the 48-hour treatment period at any tested concentrations,
. a decrease in cell viability (i.e.cell viability < 70%) was noted at concentrations = 31.4 µM,
. the corresponding IC30 and IC50 were calculated to be 26.99 and 28.31 µM, respectively,
. statistically significant gene-fold inductions above the threshold of 1.5 were noted in comparison to the negative control at 15.8 and 22.3 µM with an apparent overall dose-response relationship up to the cytotoxic concentrations,
. the Imax was 3.97 and the calculated EC1.5 was 13.72 µM.
Thus, the results met the criteria of a positive response.
Global analysis from both runs:
The geometric means IC30and IC50of the twovalidated runs were 17.68 and 18.98 µM, respectively.
The evaluation criteria for a positive response were met in both runs, the final outcome is therefore positive. This positive result can be used to support the discrimination between skin sensitizers and non-sensitizers in the context of an integrated approach to testing and assessment.
Discussion
Under the experimental conditions of this study, the test item was positive in the KeratinoSens assay. However, it is worthy to note that the two major constituents of the test item are classified H314 "causes severe skin burns and eye damage" according to the classification provided by companies to ECHA inREACH registrations (information specified by the Sponsor in an email dated 31 March 2017 archived with the study files).
According to the EURL ECVAM Recommendation on the KeratinoSens assay for skin sensitization testing (European Commission - February 2014 (JRC 87551)), reactive chemicals that cause dermal corrosion/irritation without, however, being skin sensitizers, may lead to false positive results in the KeratinoSens test method.
Thus, in the context of an integrated approach to testing and assessment, this information should be considered and the result of this test be taken with caution.
Conclusion
Under the experimental conditions of this study, the test item was positive in the KeratinoSens assay and therefore was considered to activate the Nrf2 transcription factor, though with caution due to the H314 classification (“causes severe skin burns and eye damage”) of the two major constituents of the test item.
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.