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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12-06-2013 to 17-06-2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: study performed according to the guideline and under GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Principles of method if other than guideline:
exposure period 15 minutes with post-exposure incubation period of 42 hours
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: gel
Details on test material:
- Substance type: UVCB
- Physical state: paste
- Purity: 100% (UVCB)
- Storage condition of test material: room temperature in dark

In vitro test system

Test system:
human skin model
Control samples:
yes, concurrent negative control

Test system

Details on study design:
The EpiSKIN assay uses reconstructed human epidermis to assess skin irritation. For the viability test enzymatic conversion of MTT to formazan is used.

In a pre-test direct interaction of the test substance with the detection chemical MTT is assessed.
In the main study triplicate tissues were treated with 30 uL of the test substance during 15 minutes. In addition a negative control (DPBS) and a positive control (5% SDS) were included. After treatment tissues were rinsed, incubated in maintenance fluid for 42 hours at 37 °C and treated with MTT during 3 hours (at 37 °C). After MTT loading a total biopsy of each epidermis (epidermis and collagen) was made, that was suspended in acidified iso-propanol for 3 days and kept in the refrigerator at 1-10°C. Thereafter the optical density of the extracted MTT solutions was measured at 562 nm. Viability was expressed as percentage MTT conversion versus negative control.
Tissue viability > 50% means non-irritant
Tissue viability ≤ 50% means irritant

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Value:
81.8
Negative controls validity:
valid
Positive controls validity:
valid
Remarks:
rel viability 10.6%

In vivo

Other effects:
The test substance did not interfere with the reduction of MTT to blue formazan salt.

Any other information on results incl. tables

 

Mean OD562 ± SD

Relative mean viability ± SD

Negative control:

0.946 ± 0.119

100 ± 12.6%

Test substance:

0.774 ± 0.021

81.8 ± 2.2%

SDS:

0.100 ± 0.014

10.6 ± 1.5%

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The test substance is non-irritant in the EPISKIN reconstructed human epidermal model.
Criteria used for interpretation of results: other: tissue viability > 50%: non-irritant
Executive summary:

In this assay triplicate reconstructed human skin tissues were treated with 30 uL of the test substance during 15 minutes. After treatment tissues were rinsed, kept for 42 hours and treated with MTT. Optical density of extracted MTT solutions was measured at 562 nm. Viability was expressed as percentage MTT conversion versus negative control. The viability was 81.8%, which is indicative for non-irritant substances.

The test substance is non-irritant in the EPISKIN reconstructed human epidermal model.