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EC number: 932-165-8 | 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
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Eye irritation
Administrative data
- Endpoint:
- eye irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 25 Jul - 06 Sep 2018
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 019
- Report date:
- 2019
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)
- Version / remarks:
- adopted 25 Jun 2018
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Schwabach, Germany
Test material
- Reference substance name:
- Vinasses, residue of fermentation containing biomass of bakers yeast, salt-enriched
- EC Number:
- 932-165-8
- Molecular formula:
- Not applicable (a generic molecular formula cannot be provided for this specific UVCB substance).
- IUPAC Name:
- Vinasses, residue of fermentation containing biomass of bakers yeast, salt-enriched
Constituent 1
- Specific details on test material used for the study:
- STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: sterilization at 121 °C for 20 min to avoid contaminations of the cell culture.
Test animals / tissue source
- Species:
- human
- Strain:
- other: EpiOcularTM, reconstructed three-dimensional human corneal epithelium
Test system
- Vehicle:
- unchanged (no vehicle)
- Controls:
- yes, concurrent positive control
- yes, concurrent negative control
- Amount / concentration applied:
- TEST MATERIAL
- Amount applied: 50 µL (83.3 µL / cm²)
NEGATIVE CONTROL:
- Amount applied: 50 µL
- Lot No.: RNBG3520
POSITIVE CONTROL:
- Amount applied: 50 µL
- Lot No.: S6943111 - Duration of treatment / exposure:
- 30 ± 2 min at 37 ± 2 °C
- Duration of post- treatment incubation (in vitro):
- 120 ± 15 min at 37 ± 2 °C
- Number of animals or in vitro replicates:
- 2 tissues for each treatment and control group
- Details on study design:
- DETAILS OF THE TEST PROCEDURE USED
The EpiOcular™ Eye Irritation Test (EIT) consists of a topical exposure of the neat test item to a human reconstructed cornea model followed by a cell viability test. Cell viability is measured by dehydrogenase conversion of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide], present in cell mitochondria, into a blue formazan salt that is quantitatively measured after extraction from tissues. The percent reduction of cell viability in comparison of untreated negative controls is used to predict eye irritation potential.
RHCE TISSUE CONSTRUCT
- Model used: OCL-200-EIT (MatTek Corporation, Bratislava, Slovakia)
- Tissue Batch number: 27062 (main experiment), 27066 (viable tissue controls), 27027 (killed tissue controls negative control), 27056 (killed tissue control, test item treated)
FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: The quality of the EpiOcular™ tissues was assessed via MTT cytotoxicity assay. The determined OD (540-570 nm) was in the accepted range of 1.0 - 3.0.
- Barrier function: The barrier function was evaluated by determination of the exposure time required to reduce tissue viability by 50% (ET-50) upon treatment with 100 µL of 0.3% Triton X-100. The ET-50 value fits to the acceptance criteria of 12.2 – 37.5 min.
- Contamination: The cells used to produce the EpiOcularTM tissue were screened for the presence of viruses, bacteria, yeast and other fungi. No contamination was detected. Tissue sterility was confirmed by long-term incubation with antibiotic and antimycotic free culture.
DURATION AND TEMPERATURE
- EXPOSURE: 30 ± 2 min at 37 ± 2 °C
- POST-EXPOSURE IMMERSION: 12 ± 2 min at room temperature
- POST-EXPOSURE INCUBATION: 120 ± 15 min at 37 ± 2 °C
REMOVAL OF TEST MATERIAL AND CONTROLS: The tissues were extensively rinsed with Dulbecco's phosphate buffered saline (DPBS).
INDICATION OF CONTROLS USED FOR DIRECT MTT-REDUCERS AND/OR COLOURING TEST CHEMICALS
The ability of the test substance to directly reduce MTT and to form a blue/purple reaction product was assessed in a pre-experiment. Since the MTT solution colour turned black to green, the test substance is presumed to have reduced the MTT. Therefore, an additional test using freeze-killed tissues was performed. A second pre-test revealed that the test item was also colour interfering upon mixing with water. Therefore coloured tissue controls were performed using two additional viable tissues. Moreover, since the test item showed non-specific colouring of living tissues, a third control for non-specific tissue colour in killed tissues was performed to avoid a possible double-correction for colour interference. The true tissue viability was then calculated as the percent living tissue viability obtained with living tissues minus NSMTT minus NSCliving plus NSCkilled.
True Tissue Viability = [%] mean Tissue viability - NSMTT - NSCliving + NSCkilled
NSMTT: non-specific reduction of MTT
NSCliving: non-specific colour of additional viable tissues
NSCkilled: non-specific colour of additional killed tissues
- No. of replicates: 2 tissues (killed/viable) for each test item and controls for determination of NSMTT, NSCliving and NSCkilled.
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 h ± 15 min at 37 ± 2 °C
- Spectrophotometer: plate spectrophotometer
- Wavelength: 570 nm
- Filter band pass: ± 30nm
EVALUATION CRITERIA
The test substance was considered to be not irritating to eye if the tissue viability after 30 min exposure, 12 min post-exposure immersion and 120 min post-exposure incubation is > 60% relative to the negative control treated tissue.
TEST ACCEPTANCE CRITERIA
- Negative control: The mean absolute OD 570 nm is between 0.8 and 2.5.
- Positive control: The mean relative tissue viability is < 50% of the negative control viability.
- Relative tissue viability difference of replicate tissues is < 20%.
REFERENCE TO HISTORICAL DATA OF THE RHCE TISSUE CONSTRUCT: Please refer to Table 1
Results and discussion
In vitro
Results
- Irritation parameter:
- other: % tissue viability mean value of 2 tissues
- Run / experiment:
- 30 min exposure
- Value:
- 97.1
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other:
- Remarks:
- tissue viability was corrected for non-specific reduction of MTT and non-specific colour interference
- Other effects / acceptance of results:
- ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD of the tissue replicates treated with the negative control was ≥ 0.8 and ≤ 2.8 (1.961).
- Acceptance criteria met for positive control: The mean viability of the tissue replicates treated with the positive control was <50% compared to the negative control (31.1%).
- Relative tissue viability difference of replicate tissues was < 20% (values between 1.5 and 9.7).
Any other information on results incl. tables
Table 2: Results of the test item and controls
Name |
Negative Control |
Positive Control |
Test item |
|||
Tissue |
1 |
2 |
1 |
2 |
1 |
2 |
OD570values |
1.950 |
1.957 |
0.553 |
0.737 |
1.950 |
1.902 |
1.944 |
1.995 |
0.543 |
0.732 |
1.894 |
1.866 |
|
OD570values |
1.905 |
1.911 |
0.508 |
0.692 |
1.905 |
1.857 |
1.899 |
1.950 |
0.498 |
0.687 |
1.848 |
1.820 |
|
mean of the duplicates |
1.902 |
1.931 |
0.503 |
0.689 |
1.877 |
1.839 |
mean OD |
1.916* |
0.596 |
1.858 |
|||
TODTT- NSMTT |
- |
- |
1.860 |
|||
TODTTNSMTT and NSCliving |
- |
- |
1.856 |
|||
tissue viability [%] |
99.2 |
100.8 |
26.3 |
36.0 |
97.9 |
95.9 |
relative tissue viability difference [%] |
1.5 |
9.7 |
2.0 |
|||
mean tissue viability [%] |
100.0 |
31.1 |
96.9 |
|||
mean tissue viability [%] |
- |
- |
97.0 |
|||
mean tissue viability [%] |
- |
- |
96.8 |
|||
True Tissue Viability |
- |
- |
97.1 |
* Corrected mean OD570 of the negative control corresponds to 100% absolute tissue viability
OD: optical density
NSMTT: non-specific reduction of MTT
NSC living: non-specific color of additional viable tissues
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:
- CLP: not irritating
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