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EC number: 947-912-3 | CAS number: -
Toxicological information
Skin irritation / corrosion
Administrative data
- Endpoint:
- skin irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- 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 guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
- Version / remarks:
- adopted 18 Jun. 2019
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: Comission Regulation (EU) No. 640/2012 amending Regulation (EC) No. 761/2009, Annex III, EU method B.46 'In vitro skin irritation: reconstructed human epidermis model test'
- Version / remarks:
- adopted 06. Jul. 2012
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Landesamt für Umwelt, Mainz, Germany
Test material
- Reference substance name:
- 6-[(9Z)-octadec-9-enoyloxy]hexyl (9Z)-octadec-9-enoate; 6-hydroxyhexyl (9Z)-octadec-9-enoate
- EC Number:
- 947-912-3
- Molecular formula:
- Not applicable for UVCB
- IUPAC Name:
- 6-[(9Z)-octadec-9-enoyloxy]hexyl (9Z)-octadec-9-enoate; 6-hydroxyhexyl (9Z)-octadec-9-enoate
1
In vitro test system
- Test system:
- human skin model
- Source species:
- human
- Cell type:
- non-transformed keratinocytes
- Cell source:
- other: EpiDermTM (EPI-200-SIT)
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™(EPI-200-SIT) (MatTek In Vitro Life Science Laboratories, Bratislava)
- Tissue batch number(s): 30808
- Delivery date: 23. Jul. 2019
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1°C
- Temperature of post-treatment incubation (if applicable): 37 ± 1°C
REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: The inserts were removed from the plates using sterile forceps and rinsed immediately in 1-minute-intervals.
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 h
- Spectrophotometer: M icrotiter plate photometer Anthos Reader 2010 Flexi, Anthos Microsysteme GmbH
- Wavelength: 570 nm
FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: The quality of the EpiDerm™ tissue was assessed by an MTT cell viability test. The determined OD (540 - 570 nm) was 1.841 ± 0.032 (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) upon application of 100 µL of 1% Triton X-100. The ET-50 value was determined to be 6.26 h (acceptance criteria: 4.77-8.72 h).
- Contamination: The cells used to produce EpiDerm™ tissue are screened for potential biological contaminants. Tests for the following potential biological contaminant were performed: HIV-1 virus (Oligonucleotide-direct amplification), Hepatitis B virus (Oligonucleotide-direct amplification), Hepatitis C virus (Oligonucleotide-direct amplification), Bacteria, yeast, and other fungi (long term antibiotic, antimycotic free culture); no contaminations detected.
- Reproducibility: given
CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Direct MTT reduction by the test item had not taken place and no data correction was necessary.
NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1 experiment
PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive / irritant to skin if the tissue viability after 1 h is equal or less than 50%
- The test substance is considered to be non-irritant to skin if the tissue viability after 1 h exposure is greater than 50% - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- TEST MATERIAL
- Amount(s) applied: 30 µL
NEGATIVE CONTROL
- Amount(s) applied: 30 µL
POSITIVE CONTROL
- Amount(s) applied: 30 µL
- Concentration (if solution): 5% - Duration of treatment / exposure:
- 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:
- 1 h exposure
- Value:
- 106.4
- 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: Direct MTT reduction by the test item had not taken place and no data correction was necessary.
- Colour interference with MTT: The solution was colourless, therefore no binding capacity had to be tested.
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 for every exposure time (values between 1.429 and 1.658).
- Acceptance criteria met for positive control: The mean viability of the tissue replicates treated with the positive control for 1 hour was ≤ 20% compared to the negative control (3.1%).
- Acceptance criteria met for variability between replicate measurements: The SD of mean viability of the tissue replicates is ≤ 18% (values between 0.1% and 9.8%).
- Range of historical values if different from the ones specified in the test guideline: The mean OD of the tissue replicates treated with the negative control of all experiments up to 03. Jul. 2019 was 0.476 - 2.471 (mean: 1.787, SD 0.317).
Any other information on results incl. tables
Table 1: MTT assay after 60 min exposure
| Negative control | Positive control | Test item | |||||||
| Tissue sample | 1 | 2 | 3 | 1 | 2 | 3 | 1 | 2 | 3 |
| OD570 | 1.585 | 1.702 | 1.443 | 0.080 | 0.081 | 0.082 | 1.523 | 1.770 | 1.752 |
| 1.586 | 1.682 | 1.482 | 0.080 | 0.081 | 0.084 | 1.519 | 1.760 | 1.749 | |
| OD570(mean-blank) | 1.552 | 1.658 | 1.429 | 0.046 | 0.047 | 0.049 | 1.487 | 1.731 | 1.717 |
| OD570(mean of 3 tissues) | 1.546 | 0.047 | 1.645 | ||||||
| Viability (%) | 100 | 3.1 | 106.4 | ||||||
| SD | - | 0.1 | 8.9 | ||||||
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
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