Registration Dossier

Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
29 July, 2016
Deviations:
no
Remarks:
No deviations occurred that impacted the study integrity or interpretation of the results.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch 20160912
- Expiration date of the lot/batch: 01 August 2018
- Purity test date: 25 July 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Stable until 01 August 2018
- Stability under test conditions: Stable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test article was applied neat.

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
other: Normal, human-derived epidermal keratinocytes that have been cultured to form a multilayered, highly differentiated model of the human epidermis.
Cell source:
other: MatTek Keratinocyte strain: 4F1188
Source strain:
other: Human
Details on animal used as source of test system:
SOURCE ANIMAL
- Source: Human: MatTek Keratinocyte strain: 4F1188
Justification for test system used:
Test system recommended by OECD Guideline 431.
Vehicle:
unchanged (no vehicle)
Details on test system:
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 C
- Temperature of post-treatment incubation (if applicable): 37 C
REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps: No data
- Observable damage in the tissue due to washing: No data
DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Spectrophotometer: TECAN Infinite M200 Pro Plate Reader
- Wavelength: 570 nm
- Filter: No data
- Filter bandwidth: No data
NUMBER OF INDEPENDENT TESTING RUNS / EXPERIMENTS TO DERIVE FINAL PREDICTION: 3
PREDICTION MODEL / DECISION CRITERIA
The in vitro skin corrosion test is considered acceptable if it meets the following criteria:
a) The absolute mean OD570 of the two tissues of the negative control should reasonably be
within the laboratory historical control data range.
b) The mean relative tissue viability following 1-hour exposure to the positive control
should be <15 %.
c) In the range 20 - 100% viability, the Coefficient of Variation (CV) between tissue
replicates should be less than or equal to 30%.
d) The %NSC should be less than or equal to 30% relative to the negative control OD.
e) The non-specific MTT reduction should be less than or equal to 30% relative to the negative control OD.
A test item is considered corrosive in the in vitro skin corrosion test if:
a) The relative mean tissue viability obtained after 3-minute treatment compared to the
negative control tissues is decreased below 50%.
b) In addition, a test item considered non-corrosive (viability 50%) after the 3-minute
treatment is considered corrosive if the relative tissue viability after 1-hour treatment with
the test item is decreased below 15%.
A test item is considered non corrosive in the in vitro skin corrosion test if:
a) The relative mean tissue viability obtained after the 3-minute treatment compared to the
negative control tissues is not decreased below 50%.
b) In addition, the relative tissue viability after the 1-hour treatment is not decreased below
15%.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 uL

VEHICLE: None

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): Milli-Q water: 50 uL

POSITIVE CONTROL
- Amount(s) applied (volume or weight): Potassium hydroxide: 50 uL
- Concentration (if solution): 8N
Duration of treatment / exposure:
3 minutes, 1 hour
Duration of post-treatment incubation (if applicable):
3 hours
Number of replicates:
4

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3 minute exposure
Value:
ca. 90
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1 hour exposure
Value:
ca. 6.3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other:
Remarks:
Tissue viability after 1-hour exposure is less than 15%, indicating that the test article is corrosive.
Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system: No data
- Direct-MTT reduction: The test article does not directly reduce MTT
- Colour interference with MTT: None

DEMONSTRATION OF TECHNICAL PROFICIENCY: Yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Acceptance criteria met for variability between replicate measurements: Yes

Applicant's summary and conclusion

Interpretation of results:
Category 1B (corrosive) based on GHS criteria
Conclusions:
Based on the results of the study, the test article is corrosive to the skin.
Executive summary:

The skin corrosion potential of MTDID 49220 was evaluated through topical application on a reconstructed human epidermal model for 3 minutes and 1 hour exposures. The study was conducted according to OECD 431 (2017) and was conducted in compliance with OECD GLP. Batch 20160912 of MTDID 49220 was a clear yellow liquid. MTDID 49220 was applied undiluted (50 μl) directly on top of the skin tissue. Negative (Milli-Q water) and positive (8N Potassium hydroxide) controls were tested concurrently. The relative mean tissue viability for test article-treated tissues obtained after 3-minute and 1-hour treatments with MTDID 49220 compared to the negative control tissues was 90% and 6.3%, respectively. Based on the results of the study, the test article is corrosive to the skin.