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EC number: 241-443-1 | CAS number: 17418-59-6
- 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
![](https://echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Skin irritation / corrosion
Administrative data
- Endpoint:
- skin irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 03 June 2021 to 05 June 2021
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 021
- Report date:
- 2021
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
- Version / remarks:
- 2008
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
- Version / remarks:
- 2020
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: EURL ECVAM DB-ALM Protocol n° 131 (09 June 2012): EpiSkinTM Skin Irritation Test 15 min – 42 hours
- Version / remarks:
- 2012
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- 1-amino-4-hydroxy-2-(2-phenoxyethoxy)anthraquinone
- EC Number:
- 241-443-1
- EC Name:
- 1-amino-4-hydroxy-2-(2-phenoxyethoxy)anthraquinone
- Cas Number:
- 17418-59-6
- Molecular formula:
- C22H17NO5
- IUPAC Name:
- 1-amino-4-hydroxy-2-(2-phenoxyethoxy)-9,10-dihydroanthracene-9,10-dione
- Test material form:
- solid: particulate/powder
- Details on test material:
- Physical state: Red powder
Storage condition: at room temperature
Constituent 1
In vitro test system
- Test system:
- human skin model
- Remarks:
- EpiSkin™ Small Model (EpiSkin™SM)
- Source species:
- human
- Cell type:
- non-transformed keratinocytes
- Cell source:
- other: See below
- Source strain:
- other: Adult human derived
- Details on animal used as source of test system:
- EpiSkinTM Small Model (EpiSkinTMSM), manufactured by EPISKIN Laboratories Lyon, France, is a three-dimensional human epidermis model. Adult human-derived epidermal keratinocytes are seeded on a dermal substitute consisting of a collagen type I matrix coated with type IV collagen. A highly differentiated and stratified epidermis model is obtained after 13-day culture period comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum (Tinois et al., 1994). Its use for skin irritation testing involves topical application of test materials to the surface of the epidermis, and the subsequent assessment of their effects on cell viability.
Supplier: EPISKIN Laboratories 4, rue Alexander Fleming, 69366 Lyon Cedex 07 - France
Batch No.: 21-EKIN-022
Expiry date: 07 June 2021 - Justification for test system used:
- The EPISKIN model has been validated for irritation testing in an international trial. After a review of scientific reports and peer reviewed publications on the EPISKIN method, it showed evidence of being a reliable and relevant stand-alone test for predicting rabbit skin irritation, when the endpoint is evaluated by MTT reduction and for being used as a replacement for the Draize Skin Irritation test (OECD TG 404 and Method B.4 of Annex V to Directive 67/548/EEC) for the purposes of distinguishing between skin irritating and no- skin irritating test substances (STATEMENT ON THE VALIDITY OF IN-VITRO TESTS FOR SKIN IRRITATION; ECVAM; Institute for Health & Consumer Protection; Joint Research Centre; European Commission; Ispra; 27 April 2007).
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiSkinTM Small Model
- Tissue batch number(s): 21-EKIN-022
- Production date: not specified
- Shipping date: not specified
- Delivery date: not specified
- Date of initiation of testing: 03 June 2021
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature (22.9-23.7 °C)
- Temperature of post-treatment incubation (if applicable): 37±1 °C
NUMBER OF REPLICATE TISSUES: Three replicates were used for the test item and positive and negative controls.
Additionally, 2 replicates of colour controls (NSCliving), 2 replicates of non-specific colour control (NSCkilled), 3 killed test item treated tissues and 3 killed negative control treated tissues were used for the MTT evaluation.
REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: 25 mL 1x PBS (phosphate buffered saline) solution
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 mg/mL
- Incubation time: 3 hours (± 5 min)
- Spectrophotometer: Varioskan™ LUX Type 3020
- Wavelength: 570 nm
CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Fresh tissues / killed tissues
- Procedure used to prepare the killed tissues (if applicable): Place the living epidermis in a 12 well plate with 2 mL of distilled water (replacing the culture medium). Incubate at 37±1 °C in an incubator with 5±1 % CO2, ≥ 95 % humidified atmosphere for 48 h +/- 1 hours. At the end of the incubation, discard the water.
- N. of replicates : 3 killed test item treated tissues and 3 killed negative control treated tissues
- Method of calculation used:
Non-specific MTT reduction calculation (NSMTT):
NSMTT = [(ODKT - ODKNC) / ODNC] × 100
ODKNC: negative control treated killed tissues OD (mean)
ODKT: test item treated killed tissues OD (mean)
ODNC: negative control OD (mean)
NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 5 (Test Item, Positive and negative control, Additional controls for MTT direct interacting chemicals, Additional controls for dyes and chemicals able to colour the tissue and Additional controls for non-specific colour in killed tissues)
PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive or irritating to skin if the viability after 15 minutes exposure is less than 50% of the negative control - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- yes, concurrent MTT non-specific colour control
- other: yes, for the non-specific OD evaluation (NSCliving), and also to detect and correct for test item interference with the viability measurement
- Amount/concentration applied:
- TEST MATERIAL
- Amount(s) applied: 10 mg
NEGATIVE CONTROL
- Amount(s) applied: 10 μL (1x PBS)
POSITIVE CONTROL
- Amount(s) applied: 10 μL (SDS 5%) - Duration of treatment / exposure:
- 15 minutes (± 0.5 min)
- Duration of post-treatment incubation (if applicable):
- 42 hours (± 1 h)
- Number of replicates:
- In this assay 3 replicates of test item, 3 replicates of negative control, 3 replicates of positive control, 2 replicates of colour controls (NSCliving), 2 replicates of non-specific colour control (NSCkilled), 3 killed test item treated tissues and 3 killed negative control treated tissues were used for the MTT evaluation.
Results and discussion
In vitro
Resultsopen allclose all
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- Mean of 3 runs
- Value:
- 75
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- % tissue viability
- Remarks:
- corrected relative viability
- Run / experiment:
- Mean of 3 rubs
- Value:
- 71
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Other effects / acceptance of results:
- - OTHER EFFECTS:
- Visible damage on test system: Not specified
- Direct-MTT reduction: Not significant
- Colour interference with MTT: Not significant
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes. The mean OD value of the three negative control tissues was 1.310 and the % tissue viability was 100%.
- Acceptance criteria met for positive control: Yes. The mean OD value obtained for the positive control was 0.059 and this result corresponds to 4 % viability.
- Acceptance criteria met for variability between replicate measurements: Yes. Each calculated standard deviation value (SD) for the % viability was below 18. All validity criteria were within acceptable limits and therefore the study can be considered as valid.
Any other information on results incl. tables
OD values and viability percentages of the positive and negative controls and test material
Substance | Optical Density (OD) | TODTT | Viability (%) | Relative Viability (%) | |
Negative control: 1x PBS | 1 | 1.408 | - | 108 | - |
2 | 1.311 | - | 100 | - | |
3 | 1.210 | - | 92 | - | |
mean | 1.310 | - | 100 | - | |
standard deviation (SD) | 7.57 | - | |||
Positive Control: SDS (5 % aq.) | 1 | 0.049 | - | 4 | - |
2 | 0.048 | - | 4 | - | |
3 | 0.079 | - | 6 | - | |
mean | 0.059 | - | 4 | - | |
standard deviation (SD) | 1.32 | - | |||
Test Material | 1 | 0.973 | 0.918 | 74 | 70 |
2 | 1.158 | 1.102 | 88 | 84 | |
3 | 0.826 | 0.771 | 63 | 59 | |
mean | 0.986 | 0.930 | 75 | 71 | |
standard deviation (SD) | 12.69 | 12.69 |
INDICATOR FOR POTENTIAL FALSE VIABILITY
Possible direct MTT reduction with test material - As the test material has an intrinsic colour (red), the check-method for possible direct MTT reduction with test material was not completed for the following reason: if the test material has dark colour (red, green, blue ect.), the additional controls were used automatically. This step prevents the false viability which is the possible interaction with MTT, and it occurs if the test material cannot be removed totally from the surface of the tissues. In this situation the direct interaction with MTT is defined based on the OD results of additional controls. The non-specific MTT reduction (NSMTT) was determined to be 4 %. As the NSMTT was below 50 %, the true MTT metabolic conversion and the correction of viability percentages were undertaken.
Colouring potential of test material - The test material has an intrinsic colour (red). If the test material is not white, off white, almost white and/or colourless the additional controls are automatically used and based on the OD results of additional controls the Non Specific Colour % (NSCliving %) is determined. This step prevents the false viability, which can possibly be caused by the stained surface of the tissues by the test material. Two additional test material-treated tissues were used for the non-specific OD evaluation. Mean OD (measured at 570 nm) of these tissues was determined as 0.046. The Non Specific Colour % (NSCliving %) was calculated as 3 % (below 5 %). Therefore, additional data calculation was not necessary. A false estimation of viability can be precluded.
Applicant's summary and conclusion
- Interpretation of results:
- other: Not classified in accordance with EU criteria
- Conclusions:
- In this in vitro skin irritation test using the EPISKIN model, the test material did not show significantly reduced cell viability in comparison to the negative control (mean corrected relative viability value: 71 %). All obtained test material viability results were above 50 % when compared to the viability values obtained from the negative control. Therefore, the test material was considered to be non-irritant to skin under the conditions of this study.
- Executive summary:
The skin irritation potential of the test material was investigated in accordance with the standardised guidelines OECD 439 and EU Method B. 46 in accordance with GLP using the EPISKIN Model.
Using this method, a small amount of the test material was applied to the surface of a three-dimensional human epidermis model (EpiSkin) and the optical density (OD) measured after an incubation period, from which the relative cell viability could be calculated. Additionally, positive and negative controls were performed, as well as controls for possible MTT-interacting items and chemicals able to colour the tissue.
Since the % relative tissue viability was calculated to be >50% for the test item, under the conditions of this study the test material was not irritating.
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.
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