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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
23 October 2017 - 07 February 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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
2016
Deviations:
no
Qualifier:
according to
Guideline:
other: EU Methode B.40 bis (In vitro Skin Corrosion: Human Skin Model Test)
Version / remarks:
2008
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: Reconstructed Human Epidermis (RHE)
Justification for test system used:
The use of the EpiDerm Kit is state of the art.
Vehicle:
water
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDermTM (EPI-212-SCT) from MatTek In Vitro Life Science Laboratories, Bratislava
- Tissue batch number(s): 25850
- Delivery date: 24 Oct. 2017
- Date of initiation of testing: 26 Oct. 2017

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 °C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: thoroughly rinsed with DPBS
- Modifications to validated SOP: no

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 h
- Spectrophotometer: Anthos Reader 2010 Flexi (Anthos Microsysteme GmbH)
- Wavelength: 570 nm

NUMBER OF REPLICATE TISSUES: 2

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50%, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15%.
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is greater than or equal to 15%.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount applied to the tissues: ca. 25 mg

NEGATIVE CONTROL
- Amount applied: 50 µL water

POSITIVE CONTROL
- Amount applied: 50 µL potassium hydroxide
- Concentration: 8 M
Duration of treatment / exposure:
3 and 60 minutes
Number of replicates:
2 tissues per exposure period, 3 replicates each

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
exposure period 3 minutes
Value:
72.8
Vehicle controls valid:
not examined
Negative controls valid:
yes
Positive controls valid:
yes
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
exposure period 60 minutes
Value:
19.3
Vehicle controls valid:
not examined
Negative controls valid:
yes
Positive controls valid:
yes
Other effects / acceptance of results:
- OTHER EFFECTS:
- Direct-MTT reduction: No
- Colour interference with MTT: No

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:
other: not Category 1 (corrosive) based on GHS criteria
Conclusions:
The test item is considered to be non-corrosive to skin.
Executive summary:

The test substance was tested for skin corrosion in vitro using the EpiDermTM test kit according to the OECD guideline 431 under GLP conditions. In brief, reconstructed human epidermis was exposed to either water (negative control), 8 N potassium hydroxide (positive control), or the test material (25 mg with 25 µL distilled water) for 3 and for 60 minutes. Three replicates were run per dose and exposure period. Thereafter, the tissues were incubated with MTT medium. MTT is reduced to a blue formazan dye which is extracted and measured at 570 nm as a parameter for cell viability.

Following 3 minutes treatment with the test item, the mean value of relative tissue viability was reduced to 72.8 %.This value is above the threshold for corrosion potential (50%). After 1 hour treatment, mean value of relative tissue viability was reduced to 19.3 %.This value, too, is above the threshold for corrosion potential (15%). Therefore, the test item is considered to be non-corrosive to skin in the Reconstructed Human Epidermis (RhE) Test Method.