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EC number: 931-700-2 | CAS number: 66455-29-6
Skin irritation / corrosion
Administrative data
- Endpoint:
- skin corrosion: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 11 February 2019 - 14 February 2019
- 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 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Betaines, C12-14 (even numbered)-alkyldimethyl
- EC Number:
- 931-700-2
- Cas Number:
- 66455-29-6
- Molecular formula:
- C16H33NO2 C18H37NO2
- IUPAC Name:
- Betaines, C12-14 (even numbered)-alkyldimethyl
- Test material form:
- solid
- Remarks:
- The test item was prepared by drying of the manufactured liquid alkyldimethyl betaine substance.
Constituent 1
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: BLBL016270
- Expiration date of the lot/batch: 10 January 2020
- Purity test date: Not specified
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/vehicle: Soluble in water and ethanol
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not specified
In vitro test system
- Test system:
- human skin model
- Source species:
- human
- Cell type:
- non-transformed keratinocytes
- Cell source:
- other: Normal Human Epidermal Keratinocytes (NHEKs) are isolated from the epidermis of neonatal foreskin (NHEK-CRY-NEO) or adult (breast or abdominal) skin (NHEK-CRY-AD) and cryopreserved.
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDermTM (EPI-200) Reconstructed Human Epidermis
- Tissue batch number(s): 28683
- Delivery date: 12 February 2019
- Date of initiation of testing: 13 February 2019
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37°C
REMOVAL OF TEST MATERIAL AND CONTROLS
- Not specified
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- Incubation time: 3 minutes; 1 hour
- Spectrophotometer: BMG LabTech FluoStar Optima
- Wavelength: 570 nm
- Filter: No filter
- Linear OD range of spectrophotometer:
FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: 2.047 ± 0.057 hrs (PASS)
- Barrier function: 4.79 hrs (PASS)
- Morphology: PASS
- Contamination: No contamination reported (PASS)
NUMBER OF REPLICATE TISSUES: 3 - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- TEST MATERIAL
- Amount(s) applied: 25 mg
NEGATIVE CONTROL
- Amount(s) applied: 50 µL
POSITIVE CONTROL
- Amount(s) applied: 50 µL
- Concentration (if solution): 8N - Duration of treatment / exposure:
- 3 minutes; 1 hour
- Number of replicates:
- 3
Results and discussion
In vitro
Resultsopen allclose all
- Irritation / corrosion parameter:
- % tissue viability
- Remarks:
- Average % tissue viability in triplicate tissues
- Run / experiment:
- 3 minute endpoint
- Value:
- 100.67
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Remarks:
- Average viability 100.00%
- Positive controls validity:
- valid
- Remarks:
- Average viability 5.82%
- Irritation / corrosion parameter:
- % tissue viability
- Remarks:
- Average % tissue viability in triplicate tissues
- Run / experiment:
- 1 hour endpoint
- Value:
- 25.68
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Remarks:
- Average viability 100.00%
- Positive controls validity:
- valid
- Remarks:
- Average viability 5.75%
- Other effects / acceptance of results:
- OVERALL RESULT:
The test item neither reduced the viability to below 50% after 3 mins nor below 15% after 1h and therefore should be considered as Non-Corrosive.
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD570 of the negative control tissues must be ≥0.8. Actual values: 2.41 (3 min); 2.08 (1 hr). PASS.
- Acceptance criteria met for positive control: The mean of the positive control relative percentage viability, after 1hour exposure must be < 15% of the mean of the negative control. Actual value: 5.75. PASS.
- Acceptance criteria met for variability between replicate measurements: In the range between 20% and 100% viability, the coefficient of variation (CV) is an additional acceptance criterion. It should not exceed 0.3 (30%). PASS.
Any other information on results incl. tables
Cell viability measurements after 3 minutes of application.
3 Minute Endpoint |
NC |
PC |
TA2 |
|
Tissue 1 |
Aliquot 1 OD |
2.38 |
0.14 |
2.12 |
Aliquot 2 OD |
2.38 |
0.13 |
2.14 |
|
Tissue 2 |
Aliquot 1 OD |
2.50 |
0.22 |
2.45 |
Aliquot 2 OD |
2.57 |
0.22 |
2.49 |
|
Tissue 3 |
Aliquot 1 OD |
2.31 |
0.06 |
2.65 |
Aliquot 2 OD |
2.30 |
0.07 |
2.69 |
|
All Tissues |
Average OD |
2.41 |
0.14 |
2.42 |
Average Viability |
100.00 |
5.82 |
100.67 |
|
Average % SD |
4.01 |
2.63 |
9.25 |
|
Average % CV |
4.01 |
45.10 |
9.18 |
|
NC: negative control (H2O), PC: Positive control (KOH 8N), TA2: of Betaines, C12-14 (even numbered)-alkyldimethyl.
*CV value of the PC is above 30% (45.10%) however, as the percentage of viability is below 20%
(5.82%) acceptance criterion 3 is met.
Cell viability measurements after 1 hour of application.
1 Hour Endpoint |
NC |
PC |
TA2 |
|
Tissue 1 |
Aliquot 1 OD |
2.18 |
0.12 |
0.34 |
Aliquot 2 OD |
2.20 |
0.11 |
0.33 |
|
Tissue 2 |
Aliquot 1 OD |
2.08 |
0.21 |
0.54 |
Aliquot 2 OD |
2.03 |
0.23 |
0.55 |
|
Tissue 3 |
Aliquot 1 OD |
1.95 |
0.02 |
0.71 |
Aliquot 2 OD |
2.04 |
0.03 |
0.73 |
|
All Tissues |
Average OD |
2.08 |
0.12 |
0.53 |
Average Viability |
100.00 |
5.75 |
25.68 |
|
Average % SD |
4.01 |
3.76 |
7.62 |
|
Average % CV |
4.01 |
65.52 |
29.67 |
|
NC: negative control (H2O), PC: Positive control (KOH 8N), TA2: of Betaines, C12-14 (even numbered)-alkyldimethyl.
*CV value of the PC is above 30% (65.52%) however, as the percentage of viability is below 20%
(5.75%) acceptance criterion 3 is met.
Evaluation of the results- Results were checked against the following acceptance criteria:
|
|
Actual values |
Pass/Fail |
||||||||||||
Acceptance criterion 1 |
The mean OD570 of the negative control tissues must be ≥0.8. |
3 min = 2.41 1 hr = 2.08 |
Pass |
||||||||||||
Acceptance criterion 2 |
The mean of the positive control relative % viability after 1 hr exposure must be <15% of the mean of the negative control.
|
5.75 |
Pass |
||||||||||||
Acceptance criterion 3 |
In the range between 20% and 100% viability, the coefficient of variation (CV) is an additional acceptance criterion.It should not exceed 0.3(30%). |
|
Pass |
*Note that the CV values of the PC after 3 min and 1hr were above 30% (45.10% and 65.52% respectively). However, as the percentage of cell viability was below 20% (5.82% and 5.75% respectively), acceptance criterion 3 was met.
Results Summary
Test item |
Test Item ID |
Viability after 3 minutes application |
Viability ≥ 50% after 3 minutes (Yes/No) |
Viability after 1h application (% to negative control) |
Viability ≥ 15% after 1 hour (Yes/ No) |
Corrosive (C) / Non corrosive (NC) |
Betaines, C12-14 (even numbered)-alkyldimethyl |
TA2 |
100.67 |
Yes |
25.68 |
Yes |
Non-Corrosive |
Applicant's summary and conclusion
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- In an in vitro skin corrosion study according to OECD TG 431 (RHE method), Betaines, C12-14 (even numbered)-alkyldimethyl was found to be non-corrosive to skin.
- Executive summary:
The potential of Betaines, C12-14 (even numbered)-alkyldimethyl to cause corrosion to skin was assessed in an in vitro study by the reconstructed human epidermis (RHE) test method, OECD TG 431. The test material, dried to solid form, was applied neat to unchanged human-derived epidermal keratinocytes for 3 minutes or 1 hour. The test item neither reduced the viability to below 50% after 3 mins nor below 15% after 1 hour and therefore should be considered as Non-Corrosive.
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