Registration Dossier
Registration Dossier
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: 700-728-5 | CAS number: 74805-60-0
- 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:
- 15 Feb - 01 Jun 2021
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 021
- Report date:
- 2021
Materials and methods
- Principles of method if other than guideline:
- - Principle of test: The Genomic Allergen Rapid Detection™ assay for hazard assessment of skin sensitizers (GARD™skin) provides binary hazard identification of skin sensitizers (i.e., UN GHS Category 1/No category). The method evaluates the transcriptional patterns of an endpoint-specific genomic biomarker signature, referred to as the GARDskin Genomic Prediction Signature (GPS) in the SenzaCell™ cell line exposed to test chemicals.
- Short description of test conditions: The test system (SenzaCell™ cells) is exposed to a non-cytotoxic concentration of the test item and concurrent negative and positive controls for 24 h at 37 °C and 5% CO2. Cells are lysed after exposure, and RNA is isolated from the lysed cells. 100 ng of RNA is used as sample input in a hybridization assay with the endpoint specific GARDskin CodeSet. Each hybridized sample is prepared on cartridge using nCounter MAX Prep Station and individual transcripts of the endpoint specific biomarker signature are quantified using nCounter MAX Digital Analyzer
- Parameters analysed / observed: the quantified transcriptional levels of the endpoint specific biomarker signature are analysed with the GARD Data Analysis Application (GDAA) software in order to determine the Decision Value (DV), which indicates whether a test item has skin sensitising potential or not. - GLP compliance:
- yes (incl. QA statement)
- Type of study:
- other: Genomin Allergen Rapid Detection™ assay for hazard assessment of skin sensitizers (GARD™skin)
- Justification for non-LLNA method:
- Not required for transported isolated intermediates <1000 t/a
Test material
- Reference substance name:
- (5Z)-5-(phenylmethylidene)imidazolidine-2,4-dione
- EC Number:
- 700-728-5
- Cas Number:
- 74805-60-0
- Molecular formula:
- C10H8N2O2
- IUPAC Name:
- (5Z)-5-(phenylmethylidene)imidazolidine-2,4-dione
- Test material form:
- solid: particulate/powder
Constituent 1
In vitro test system
- Details of test system:
- other: human myeloid leukemia-derived cell line SenzaCell™
- Vehicle / solvent control:
- DMSO
- Negative control:
- not applicable
- Positive control:
- other: p-Phenylenediamine (CAS No. 106-50-3)
Results and discussion
- Positive control results:
- The positive control substance p-Phenylenediamine (PDD) was tested at 75 µM, assigned a mean Decision Value (DV) of 8.9, and thus correctly identified as a skin sensitizer.
In vitro / in chemico
Results
- Key result
- Group:
- test chemical
- Run / experiment:
- mean
- Parameter:
- other: mean Decision Value (DV)
- Value:
- 2.13
- Cell viability:
- 98.23% (mean of three replicates)
- Vehicle controls validity:
- valid
- Remarks:
- mean DV: -2.24; mean relative cell viability: 99.70%
- Negative controls validity:
- valid
- Remarks:
- mean DV: n.a.; mean absolute cell viability: 96.90%
- Positive controls validity:
- valid
- Remarks:
- mean DV: 8.9; mean relative cell viability: 91.33%
- Remarks on result:
- positive indication of skin sensitisation
- Outcome of the prediction model:
- positive [in vitro/in chemico]
Any other information on results incl. tables
SOLUBILITY ASSESSMENT
The vehicle chosen for downstream procedures was DMSO at 0.5% in-well resulting in an in-well concentration of 500 µM of the Test Item.
Vehicle (max in-well concentration of vehicle)(I) | Maximum assessed concentration in vehicle (mM) | Solubility conclusion in vehicle(II) | Solubility in medium (µM) | Solubility conclusion in-well(II) |
DMSO (0.5%) | 500 | Soluble | 500 | Soluble with 0.5% DMSO |
(I) Maximum concentration with an accurate classification as non-sensitiser in GARDskin. Based on historical data and previous experience. Data not shown.
(II) As defined by ocular inspection.
PHENOTYPIC QUALITY CONTROL
To ensure that the cells are maintained in an inactivated state and to detect a potential phenotypic drift, a quality control of the cells was performed the day of cell stimulation. The cells passed the specified acceptance criteria in all cell stimulations performed in the Study.
CYTOTOXICITY ASSESSMENT
The Test ltem was assayed in the concentration range 55-625 µM. Since no cytotoxicity was observed, the highest permissible concentration in the GARDskin assay, 500 µM was chosen for Main stimulations.
Relative viability (%) | |||||||
In-well concentration (µM) | 55 | 83 | 123 | 185 | 278 | 415 | 625 |
Test Item | 99.6 | 99.8 | 99.7 | 99.8 | 99.8 | 98.9 | 97.6 |
MAIN STIMULATIONS
The relative viability of the Test and Reference ltems and absolute viability of the unstimulated control are presented below. All samples passed the acceptance criteria.
Test/Reference Item | Replicate# | Stimulation concentration (I) | Vehicle | Relative viability(II) (%) | Acceptance criteria (Pass/Fail) |
Test Item | 1 | 500 µM | DMSO 0.5% | 97.9 | Pass |
2 | 98.1 | ||||
3 | 98.7 | ||||
Positive control, PPD | 1 | 75 µM | DMSO 0.1% | 91.5 | |
2 | 89.8 | ||||
3 | 92.7 | ||||
Negative control, DMSO | 1 | 0.50% | N/A | 100.3 | |
2 | 99.5 | ||||
3 | 99.3 | ||||
Unstimulated control (absolute viability, %) | 1 | N/A | 96.7 | ||
2 | 96.8 | ||||
3 | 97.2 |
(I) Concentration based on max. solubility and cytotoxicity
(II) Observed viability, calculated based on the equation in section Standard GARD Assay Procedure
RNA QUALITY CONTROL
The result from the RNA Quality Control is presented below. All samples passed the acceptance criteria.
Test/Reference Item | Replicate# | RNA conc (ng/µL) | RIN | RNA QC (Pass/Fail) |
Test Item | 1 | 100 | 9.9 | Pass |
2 | 100 | 10 | ||
3 | 26 | 9.8 | ||
Positive control, PPD | 1 | 100 | 10 | |
2 | 84 | 9.7 | ||
3 | 75 | 9.8 | ||
Negative control, DMSO | 1 | 127 | 10 | |
2 | 120 | 9.9 | ||
3 | 85 | 10 | ||
Unstimulated control | 1 | 151 | 10 | |
2 | 100 | 9.4 | ||
3 | 82 | 9.6 |
ENDPOINT MEASUREMENT QUALITY CONTROL
The result from the Endpoint Measurement Quality Control is below. All samples passed the acceptance criteria.
Test/Reference Item | Replicate# | Imaging Quality | Binding Density | Limit of detection | Linearity | Enpoint measurement QA (Pass/Fail) |
Test Item | 1 | 0.97 | 0.4 | 7.8 | 0.997 | Pass |
2 | 0.97 | 0.34 | 5.5 | 0.997 | ||
3 | 0.98 | 0.34 | 6.4 | 0.997 | ||
Positive control, PPD | 1 | 0.97 | 0.33 | 6.7 | 0.998 | |
2 | 0.96 | 0.33 | 7.5 | 0.995 | ||
3 | 0.97 | 0.31 | 8.8 | 0.997 | ||
Negative control, DMSO | 1 | 0.98 | 0.43 | 6.2 | 0.996 | |
2 | 0.99 | 0.33 | 4.9 | 0.997 | ||
3 | 0.95 | 0.36 | 5.5 | 0.997 | ||
Unstimulated control | 1 | 0.98 | 0.25 | 7.1 | 0.995 | |
2 | 0.97 | 0.42 | 5.6 | 0.997 | ||
3 | 0.92 | 0.41 | 6.5 | 0.998 |
CLASSIFICATION OF TEST AND REFERENCE ITEMS
Decision Values were generated for each replicate sample by the GARDskin prediction model using the GDAA. as described in section Standard GARD Assay Procedure.
Individual Decision Values and corresponding mean Decision Values used for GARDskin classifications of Test and Reference ltems are presented below. All samples passed the acceptance criteria.
Test/Reference Item | Decision Value (DV) | Mean DV | GARDskin classification | Acceptance criteria(I) (Pass/Fail) | ||
Replicate #1 | Replicate #2 | Replicate #3 | ||||
Test Item | 2.57 | 2.63 | 1.19 | 2.13 | Sensitiser | Pass |
Positive control, PPD | 8.98 | 8.84 | 8.88 | 8.9 | Sensitiser | |
Negative control, DMSO | -1.87 | -2.28 | -2.56 | -2.24 | Non-sensitiser |
(I) Reference Item acceptance criteria as described in section Standard GARD Assay Acceptance Criteria
STATISTICAL SIGNIFICANCE OF READOUT
The conclusions presented in this Study report are based on generated output from the GARD prediction model, which is based on a Support Vector Machine. The SVM is trained on a training dataset consisting of 120 samples with measurements of 200 variables, each of which have been shown to be statistically significantly regulated by sensitizers compared to non-sensitizers (p < 5x10-20, q < 2x10-6, multiple hypothesis correction using Benjamini-Hochberg).
Applicant's summary and conclusion
- Interpretation of results:
- Category 1 (skin sensitising) based on GHS criteria
- Conclusions:
- For the GARDskin classification, the mean DV of three replicates samples for each Test and Reference Item were used. The mean DV generated for the Test Item was 2.13, and thus ≥0 (skin sensitiser). The positive and negative controls were correctly classified as skin sensitiser (mean DV: 8.9) and skin non-sensitiser (mean DV: -2.24), respectively, in the GARDskin assay.
All specified acceptance criteria passed in this study and the study was therefore valid.
In conclusion, based on the results of this study, the Test Item was classified as sensitiser in the GARDskin assay tested at an in-well concentration of 500 µM.
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.
