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Skin irritation / corrosion

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skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
30th June 2020 to 10th July 2020
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
GLP compliance:

Test material

Reference substance name:
Cas Number:
Test material form:
solid: particulate/powder

In vitro test system

Test system:
human skin model
Source species:
Cell type:
non-transformed keratinocytes
Cell source:
other: Not specified
Justification for test system used:
The protocol meets the requirements of the OECD guideline, “In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method” (TG 439)
unchanged (no vehicle)
Details on test system:
EpiDerm Skin Model
The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
- Amount applied: approximately 25 mg

Duration of treatment / exposure:
The test articles, the positive control (5% Sodium Dodecyl Sulfate (SDS)), and the negative control (Calcium & Magnesium Free-Dulbecco’s Phosphate Buffered Saline (CMF-DPBS)), were treated in triplicate EpiDermTM tissues for a 60±1 minute exposure period.
Duration of post-treatment incubation (if applicable):
42-hour post-exposure incubation
Number of replicates:

Results and discussion

In vitro

Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean of three replicates - 1hour
Negative controls validity:
Positive controls validity:
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
The assay was accepted when the following criteria were met: 1) the positive control (5% SDS) resulted in a mean tissue viability ≤ 20%, 2) the mean OD570 value of the negative control tissues was ≥ 0.8 and < 2.8, and 3) the standard deviations of the positive and negative control calculated from individual percent tissue viabilities of the three identically treated replicates were < 18%.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
According to the prediction model presented in OECD TG 439, the test article was predicted to be a GHS No Category.
Executive summary:

The EpiDerm™ Skin Model was used to assess the potential dermal irritation of the test article. The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) conversion assay, which measures the NAD(P)H-dependent microsomal enzyme reduction of MTT (and to a lesser extent, the succinate dehydrogenase reduction of MTT) to a blue formazan precipitate, was used to assess cellular metabolism after exposure to a test article 1. The protocol was based upon the OECD guideline, “In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method” (TG 439).

The test article was administered to the test system without dilution (neat). Since the test article was a powder, 25 µL of sterile Calcium & Magnesium Free Dulbecco’s Phosphate Buffered Saline (CMF-DPBS) were applied to the tissues prior to addition of approximately 25 mg of the test article, which was administered using a dosing spoon. A sterile rod was used to spread the test article over the surface of the tissues. Following a 60 ± 1 minute exposure
period, followed by a 42 ± 2-hour post-exposure incubation period, the mean percentage tissue viability was determined to be 105.8 ± 3.37. According to the prediction model presented in OECD TG 439, the test article was predicted to be a GHS No Category.