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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 812-549-0 | CAS number: 881539-89-5
- 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
Skin irritation / corrosion
Administrative data
- Endpoint:
- skin irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 19 - 22 Nov 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 020
- Report date:
- 2020
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
- Version / remarks:
- Adopted in 2019
- Deviations:
- yes
- Remarks:
- The demonstration of appropriate morphology by vendor is missing.
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Slovak National Accreditation Service, Bratislava, Slovak Republic
Test material
- Reference substance name:
- disodium 4-[(1-hydroxyprop-2-en-1-yl)amino]-4'-(prop-2-enamido)-[1,1'-biphenyl]-2,2'-disulfonate
- EC Number:
- 812-549-0
- Cas Number:
- 881539-89-5
- Molecular formula:
- C18H14N2Na2O8S2
- IUPAC Name:
- disodium 4-[(1-hydroxyprop-2-en-1-yl)amino]-4'-(prop-2-enamido)-[1,1'-biphenyl]-2,2'-disulfonate
Constituent 1
In vitro test system
- Test system:
- human skin model
- Source species:
- human
- Cell type:
- non-transformed keratinocytes
- Cell source:
- other: reconstructed human epidermal (RhE) model EpiDerm™ SIT (EPI-200), MatTek In Vitro Life Science Laboratories, Slovak Republic
- Justification for test system used:
- The test item is applied topically to a three-dimensional human reconstructed epidermis (RhE) model, comprised of non-transformed human-derived epidermal keratinocytes, which have been cultured to form a multilayered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers and a multilayered stratum corneum containing intracellular lamellar lipid layers representing main lipid classes analogous to those found in vivo. Irritant chemicals are identified by their ability to decrease cell viability (as determined by using the MTT reduction assay) below defined threshold levels. The EpiDerm™ SIT allows discrimination between substances requiring classification and labeling (EU GHS Cat. 1 or 2) and non-irritants according to the EU and GHS classification system.
- Vehicle:
- unchanged (no vehicle)
- Remarks:
- Shortly before application of the test item, the tissue surface was moistened with 25 µL sterile DPBS to improve contact of the tissue surface with the test item.
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™ SIT (EPI-200), MatTek In Vitro Life Science Laboratories, Slovak Republic
- Tissue batch number(s): Lot no. 30836, Keratinocyte strain: 00267
- Production date: 20 Nov 2019 (Certificate of Analysis from MatTek)
- Delivery date: 18 Nov 2019
- Date of initiation of testing: 19 Nov 2019
Comment on the production date and experimental starting date: The tissue supplier states that the production date for the tissues produced by MatTek IVLSL is Monday. The lab performs preQC and if the tissues do not fulfil the quality control criteria the tissues are not shipped to the customers. To obtain the real life data for the tissues shipped to the customers, MatTek mimics the shipment conditions and therefore performs the quality controls on Tuesdays with quality control measurements on Wednesdays. Therefore the certificates of analysis for the tissues are issued on Wednesdays. This is the standard procedure for RhE tissues currently produced at MatTek IVLSL.
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment/ exposure: 35 min at 37 °C + 25 min at room temperature
- Temperature of post-treatment incubation: 37 °C
REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: washed 15 times, then submerged 3 times in 150 mL Dulbecco's Phosphate-Buffered Saline (DPBS). Finally, the tissues were rinsed once from inside and once from outside with sterile DPBS and each insert was blotted on sterile blotting paper and transferred to new 6-well plates prefilled with 0.9 mL of fresh assay medium.
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT/ EXPOSURE
- MTT concentration: 1 mg/mL; 0.3 mL/well of MTT solution
- Incubation time: 3 h
- Spectrophotometer: Dynex MRX II
- Wavelength: 540 nm
- Filter: without a reference filter
FUNCTIONAL MODEL CONDITIONS
- Viability: The quality of the RhE tissue was assessed by an MTT cell viability test (4 h, n=3). The determined OD (540 - 570 nm) was 2.002 ± 0.169 (acceptance criteria: 1.0 - 3.0).
- Barrier function: The barrier function was assessed by determination of the exposure time required to reduce tissue viability by 50% (ET-50) following application of 100 µL 1% Triton X-100. Four timepoints (4, 6, 8 10 h exposure) were evaluated. The ET-50 value was determined to be 4.85 h (acceptance criteria: 4.77 - 8.72 h).
- Contamination: The cells used to produce the RhE tissue were screened for the presence of HIV-1 virus, Hepatitis B virus, Hepatitis C virus, bacteria, yeast, and other fungi. No contamination was detected.
CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
Test for interference of test substance with MTT endpoint:
Test substances may interfere with the MTT endpoint, if they are coloured and/or able to directly reduce MTT and at the same time are present in the tissues when the MTT viability test is performed. Some non-coloured test substances may change into coloured material under wet or aqueous conditions and thus stain tissues during the 60 min exposure period. To identify these possible interferences, a functional check was performed.
The test substance in amount of 25 mg was added into 0.3 mL of purified water. The mixture was incubated in glass test tube in the incubator at 37 ± 1 °C in a humidified atmosphere of 5 ± 1% CO2 in air for 60 min. At the end of the exposure time the presence of the staining was evaluated.
The colour of mixture was unchanged.
Then, 25 mg of the test substance was added to 1 mL of the MTT medium and incubated in the incubator at 37 ± 1 °C in a humidified atmosphere of 5 ± 1% CO2 in air for 60 min. Untreated MTT medium was used as control.
The colour of MTT solution remained unchanged and it was concluded that the test substance did not reduce MTT directly.
An additional functional check was not performed.
NUMBER OF INDEPENDENT TEST SEQUENCES/ EXPERIMENTS TO DERIVE FINAL PREDICTION
Single experiment
PREDICTION MODEL/ DECISION CRITERIA
- The test substance is considered to be irritant to skin if the viability after 60 min exposure is ≤ 50%.
- The test substance is considered to be non-irritant to skin if the viability after 60 min exposure is > 50%.
DEMONSTRATION OF TECHNICAL PROFICIENCY TESTING
Demonstration of proficiency is stated in amendment No. 1 to the final report. The laboratory demonstrates technical proficiency by the non-classified substances (UN GHS no Category) and classified substances (UN GHS Category 2) indicated by the OECD 439 (version 25 Jul 2015) in vitro skin irritation test. - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg
VEHICLE
- The white powder was applied unchanged using a spoon. However, in accordance with OECD TG 439, shortly before application of the test item, the tissue surface was moistened with 25 µL sterile DPBS to improve contact of the tissue surface with the test item.
NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL DPBS
POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL
- Concentration (if solution): 5% SDS - Duration of treatment / exposure:
- 60 min (35 min at 37 °C and 25 min at RT)
- Duration of post-treatment incubation (if applicable):
- 42 h
- Number of replicates:
- Triplicates for each treatment and control group.
Results and discussion
In vitro
Results
- Irritation / corrosion parameter:
- % tissue viability
- Remarks:
- mean value of 3 tissues
- Run / experiment:
- 60 min exposure
- Value:
- 87.1
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- no indication of irritation
- Other effects / acceptance of results:
- - OTHER EFFECTS:
- Direct-MTT reduction: Since the test substance did not directly reduce MTT, an additional test with freeze-killed or viable tissues was not performed.
- Colour interference with MTT: The test substance did not change the colour, when mixed with deionised water, and passed the colour interference pre-test.
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes, the tissue viability met the acceptance criterion: mean OD of negative control was 1.334.
- Acceptance criteria met for positive control: Yes, the positive control met the acceptance criterion: mean tissue viability was 4.2% (i.e. < 20%).
- Acceptance criteria met for variability between replicate measurements: Yes, SD calculated from individual % tissue viabilities of three identically treated replicates was < 18%.
See Table 1 under "Any other information on results incl. tables").
Any other information on results incl. tables
Table 1. Skin irritation potential of AF-959 after 60-minute exposure in reconstructed human epidermal model EpiDerm™ SIT
Test item | Mean of OD | SD of OD | Mean of cell viabilities (%) | SD of cell viabilities | CV (%) | Classification |
Negative control (DPBS) |
1.334 | 0.025 | 100 | 1.91 | 1.91 | non-irritant |
Positive control (5% SDS) |
0.056 | 0.002 | 4.2 | 0.15 | 3.57 | irritant |
Test substance (AF-959) |
1.162 | 0.054 | 87.1 | 4.07 | 4.68 | non-irritant |
Applicant's summary and conclusion
- Interpretation of results:
- other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.
- Conclusions:
- Under the conditions of the present study, AF-959 is considered to be non-irritant to skin (GHS No Category).
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.