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EC number: 257-854-4 | CAS number: 52333-30-9
- 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 irritation / corrosion
Administrative data
- Endpoint:
- skin irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- Experimental starting date - 02 September 2015; Experimental completion date - 07 September 2015; Study completion date - 19 December 2015.
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 015
- Report date:
- 2015
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
- Deviations:
- yes
- Remarks:
- See below
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
- Deviations:
- yes
- Remarks:
- See below
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Disodium 5,5'-[(1-methylethylidene)bis(4,1-phenyleneoxysulphonyl-2,1-phenyleneazo)]bis[6-amino-4-hydroxynaphthalene-2-sulphonate]
- EC Number:
- 257-854-4
- EC Name:
- Disodium 5,5'-[(1-methylethylidene)bis(4,1-phenyleneoxysulphonyl-2,1-phenyleneazo)]bis[6-amino-4-hydroxynaphthalene-2-sulphonate]
- Cas Number:
- 52333-30-9
- Molecular formula:
- C47H38N6O14S4.2Na
- IUPAC Name:
- disodium 5,5'-[propane-2,2-diylbis(4,1-phenyleneoxysulfonyl-2,1-phenylenediazene-2,1-diyl)]bis(6-amino-4-hydroxynaphthalene-2-sulfonate)
- Test material form:
- other: solid
Constituent 1
- Specific details on test material used for the study:
- Identification: FAT 21021/E TE
Batch: 72
Purity: ≥75 %
Physical state/ Appearance: Red solid
Expiry date:16 February 2020
Storage Conditions: room temperature in the dark.
In vitro test system
- Test system:
- human skin model
- Remarks:
- EPISKIN™ Reconstructed Human Epidermis Model
- Source species:
- human
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used:
EPISKIN Reconstructed Human Epidermis Model Kit
- Tissue batch number: 15 EKIN 035
- Date of initiation of testing:
02 September 2015.
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure:
37 °C
REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps:
One
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.2 mg/ml
- Incubation time: 3 hours
- Spectrophotometer: UV Vis
- Wavelength: 562nm
NUMBER OF REPLICATE TISSUES: Triplicate - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- Approximately 10 mg (26.3 mg/cm²) of the test item was applied
- Duration of treatment / exposure:
- 15 minutes
- Duration of post-treatment incubation (if applicable):
- 42 hours incubation at 37 °C.
- Number of replicates:
- Three
Results and discussion
In vitro
Results
- Irritation / corrosion parameter:
- % tissue viability
- Value:
- 105.1
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- no indication of irritation
- Other effects / acceptance of results:
- See "Any other information on results incl. tables"
Any other information on results incl. tables
Direct MTT Reduction
The MTT solution containing the test item did not turn blue which indicated that the test item did not directly reduce MTT.
Assessment of Color Interference with the MTT endpoint
Colored test items may interfere with the MTT endpoint. Therefore, an additional functional procedure using viable color correction tissues was performed to measure any potential interference. However, the results obtained showed that negligible interference due to the color of the test item occurred. It was therefore considered unnecessary to use the results of the color correction tissues for quantitative correction of results or for reporting purposes.
Quality Criteria
The relative mean tissue viability for the positive control treated tissues was 9.9 % relative to the negative control treated tissues and the standard deviation value of the viability was 1.1 %. The positive control acceptance criterion was therefore satisfied. The mean OD562 for the negative control treated tissues was 0.718 and the standard deviation value of the viability was 1.0 %. The negative control acceptance criterion was therefore satisfied. The standard deviation calculated from individual tissue viabilities of the three identically test item treated tissues was 1.6 %. The test item acceptance criterion was therefore satisfied.
Mean OD562Values and Percentage Viabilities for the Negative Control Item, Positive Control Item and Test Item
Item | OD562of tissues | Mean OD562of triplicate tissues | ±SD of OD562 | Relative individual tissue viability (%) | Relative mean viability (%) | ± SD of Relative mean viability (%) |
Negative Control Item | 0.726 | 0.718 | 0.007 | 101.1 | 100* | 1.0 |
0.715 | 99.6 | |||||
0.712 | 99.2 | |||||
Positive Control Item | 0.064 | 0.071 | 0.008 | 8.9 | 9.9 | 1.1 |
0.080 | 11.1 | |||||
0.069 | 9.6 | |||||
Test Item | 0.766 | 0.755 | 0.012 | 106.7 | 105.1 | 1.6 |
0.755 | 105.2 | |||||
0.743 | 103.5 |
OD = Optical Density
SD = Standard deviation
* = The mean viability of the negative control tissues is set at 100 %
Applicant's summary and conclusion
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- The test item was classified as non-irritant.
- Executive summary:
Introduction
The purpose of this test was to evaluate the skin irritation potential of the test item using the EPISKINTM reconstructed human epidermis model after a treatment period of 15 minutes followed by a post‑exposure incubation period of 42 hours. The principle of the assay was based on the measurement of cytotoxicity in reconstructed human epidermal cultures following topical exposure to the test item by means of the colorimetric MTT reduction assay. Cell viability is measured by enzymatic reduction of the yellow MTT tetrazolium salt (3‑[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to a blue formazan salt (within the mitochondria of viable cells) in the test item treated tissues relative to the negative controls.
Method
Triplicate tissues were treated with the test item for an exposure period of 15 minutes. At the end of the exposure period each tissue was rinsed before incubating for 42 hours. At the end of the post‑exposure incubation period each tissue was taken for MTT-loading. The maintenance medium from beneath each tissue was transferred to pre‑labeled micro tubes and stored in a freezer for possible inflammatory mediator determination. After MTT-loading a total biopsy of each epidermis was made and placed into micro tubes containing acidified isopropanol for extraction of formazan crystals out of the MTT‑loaded tissues. At the end of the formazan extraction period each tube was mixed thoroughly and duplicate 200 µL samples were transferred to the appropriate wells of a pre‑labeled 96‑well plate. The optical density was measured at 562 nm. Data are presented in the form of percentage viability (MTT reduction in the test item treated tissues relative to negative control tissues).
Results
The relative mean viability of the test item treated tissues was 105.1% after the 15‑Minute exposure period and 42‑Hours post‑exposure incubation period.
Quality criteria: The quality criteria required for acceptance of results in the test were satisfied.
Conclusion
The test item was classified as non-irritant. The following classification criteria apply:
EU DSD and CLP Not classified for Irritation.
UN GHS Not classified for Irritation (Category 3 cannot be determined).
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