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Diss Factsheets
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EC number: - | CAS number: -
- 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
Eye irritation
Administrative data
- Endpoint:
- eye irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 23 - 25 Sep 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 guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Hess. Ministerium für Umwelt, Klimaschutz, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany
Test material
- Reference substance name:
- 1,3-dimethylbutyl (E)-but-2-enoate
- Molecular formula:
- C10H18O2
- IUPAC Name:
- 1,3-dimethylbutyl (E)-but-2-enoate
Constituent 1
Test animals / tissue source
- Species:
- human
- Strain:
- other: EpiOcular™
- Details on test animals or tissues and environmental conditions:
- TEST MODEL (EpiOcular™ Kit)
- Source: MatTek Corporation, Ashland, USA
- Lot No.: 21572
TEST METHOD
The EpiOcular™ tissue consists of normal, human-derived epidermal keratinocytes which have been cultured to form a stratified squamous epithelium similar to that found in the human cornea. It consists of basal cells which progressively flatten out as the apical surface of the tissue is approached, analogous to the normal in vivo corneal epithelium. Irritant materials are identified by their ability to damage the underlying cell layers which is determined through a decrease in cell viability as determined by MTT reduction.
ADAPTATION TO CELL CULTURE CONDITIONS
1.0 mL assay medium (37 °C) was aliquoted into 6-well plates. The inserts with EpiOcular™ tissues were transferred aseptically into the plates and pre-incubated at standard culture conditions for 1 h. Afterwards, the medium was replaced by 1 mL fresh assay medium and the EpiOcular™ tissues were incubated at standard culture conditions overnight (18 h). After the overnight incubation, the tissues were pre-wetted with 20 µL of Ca²+ Mg²+ free DPBS. The tissues were incubated at standard culture conditions for 30 min.
INCUBATION CONDITIONS (INCUBATOR)
- Temperature (°C): 37 ± 1.5
- CO2 gas concentration (%): 5 ± 0.5
- Humidity (%): 95
Test system
- Vehicle:
- unchanged (no vehicle)
- Controls:
- yes, concurrent positive control
- yes, concurrent negative control
- Amount / concentration applied:
- TEST MATERIAL
- Applied volume: 50 µL
POSITIVE SUBSTANCE
- Substance: methyl acetate
- Applied volume: 50 µL
NEGATIVE CONTROL
- Substance: deionised water
- Applied volume: 50 µL - Duration of treatment / exposure:
- 30 min
- Duration of post- treatment incubation (in vitro):
- 120 min
- Number of animals or in vitro replicates:
- The test was performed in duplicate for each treatment and control group.
- Details on study design:
- TEST SITE
- Area of exposure: 0.6 cm²
REMOVAL OF TEST SUBSTANCE
- Washing: At the end of the treatment time, the test substance was removed by extensively rinsing the tissues with Ca²+Mg²+ free DPBS in clean beakers.
- Post-treatment incubation period: 2 h
CELL VIABILITY MEASUREMENTS
For determining alterations in cell viability, MTT reduction assays were performed. Therefore, a volume of 300 µL MTT solution was added to each well for 3 h at standard culture conditions. After removal of the MTT solution, wells were rinsed three times with Ca²+Mg²+ free DPBS. Extraction of the formazan product was carried out in 2 mL isopropanol. At the end of the extraction period the optical density (OD) was measured.
EVALUATION OF RESULTS
1) The mean OD value of the blank control wells (ODBlk) for each experiment were calculated.
2) The ODBlkfrom each OD value of the same experiment (blank corrected values) were subtracted.
3) The mean value of the two aliquots for each tissue (= corrected test substance OD) were calculated.
4) The mean value of the two relating tissues for each control and test substance (= corrected mean OD) were calculated. For further calculations only the corrected mean negative control OD value was needed.
5) The corrected OD value of the negative control corresponds to 100% viability.
6) The percent viability of each of the two relating tissues for each control and test substance relative to the negative control (100% control) were calculated.
Viability (%) = [(corrected test substance OD) / (corrected mean negative control OD)] x 100
EVALUATION CRITERIA
A test substance leading to a tissue viabilty > 60% (relative to the negative control tissue viability) is considered as non-irritating.
A test substance leading to a tissue viabilty < 60% (relative to the negative control tissue viability) is considered as irritating.
ACCEPTABILITY OF THE ASSAY
1. The negative control OD is > 0.8 and < 2.5
2. The mean relative viability of the positive control is below 60% of the negative control viability.
3. The difference of viability between the two relating tissues of a single test item is < 20% in the same run (for positive and negative control tissues and tissues of test substance). This applies also to the killed controls (substance and negative killed control) and the colorant controls which are calculated as percent values related to the viability of the relating negative control.
Results and discussion
In vitro
Results
- Irritation parameter:
- other: cell viability (%)
- Remarks:
- mean values of 2 tissues
- Run / experiment:
- 30 min exposure
- Value:
- 100.5
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Other effects / acceptance of results:
- OTHER EFFECTS:
The optical pre-experiment (colour interference pre-experiment) to investigate the test substance’s colour change potential in water or isopropanol did not led to a change in colour. Optical evaluation of the MTT-reducing capacity of the test substance with MTT-reagent did not show blue colour.
ACCEPTANCE OF RESULTS:
All acceptance criteria were met.
Any other information on results incl. tables
Table 1. Results after 30 min incubation time
Test group |
Absorbance* |
Mean absorbance of 2 tissues* |
Rel. absorbance (%)** |
Absolute value of the difference of the rel. absorbance (%) Tissue 1 and 2 |
Rel. absorbance (% of negative control)** |
||
Tissue 1 |
Tissue 2 |
Tissue 1 |
Tissue 2 |
||||
Negative control |
1.568 |
1.653 |
1.611 |
97.4 |
102.6 |
5.3 |
100.0 |
Positive control |
0.107 |
0.114 |
0.110 |
6.6 |
7.1 |
0.5 |
6.9 |
Test substance |
1.675 |
1.563 |
1.619 |
104.0 |
97.0 |
7.0 |
100.5 |
* Mean of two replicate wells after blank correction
** Relative absorbance (rounded values): 100 × (absorbance test substance/positive control) / (absorbance negative control)
Applicant's summary and conclusion
- Interpretation of results:
- other: CLP/EU GHS criteria not met, no calssification according to Regulation (EC) No 1272/2008
- Conclusions:
- Under the conditions of the conducted test, the test substance did not exhibit irritating properties towards human-derived epidermal keratinocytes in the EpiOcular™ model.
- Executive summary:
An in vitro eye irritation test using a human cornea model according to OECD 492 was conducted in compliance with GLP. Since the viability value of the test substance exposed tissues did not decrease below 60% (100.5%), the test substance is not considered to possess an eye irritating potential.
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