Registration Dossier

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2016-06-17 - 2016-08-12
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Qualifier:
according to
Guideline:
other: Skinethic skin irritation test -42bis Standard operating procedure (SOP) 2009
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Specific details on test material used for the study:
Designation: Art. 201386
Synonym: B-2O-O5
Batch: EF13004186
Purity (HPLC): 100.0%
Appearance: White, solid
Expiry date: April 30, 2018
Storage: Tightly closed, dark at room temperature (15 to 25°C)

The test item was applied neat to the tissues.

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
foreskin from a single donor
Justification for test system used:
standard model
Vehicle:
unchanged (no vehicle)
Remarks:
No vehicle used; Test item was applied neat to the tissues
Details on test system:
CELL CULTURE
- Supplier: Episkin/SkinEthic Laboratories, Lyon, France
- Source: human keratinocytes cultured on a polycarbonate filter in conditions which permit their terminal differentiation
- Format: 24 well plate
- Batch: 16-RHE-073
- Expiration date: 2016-07-18
Control samples:
yes, concurrent negative control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: 16 mg per tissue

NEGATIVE CONTROL
- Amount applied: 16 µL per tissue (Dulbecco`s Phosphate-Buffered Saline)

POSITIVE CONTROL
- Amount applied: 16 µL per tissue
- Concentration: 5% aqueous solution of sodium dodecyl sulfate in deionised water
Duration of treatment / exposure:
42 min (± 1 minute)
Duration of post-treatment incubation (if applicable):
42 hours (± 1 hour)
Number of replicates:
3

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Experiment 1 / Run 1
Value:
97.48
Vehicle controls valid:
not applicable
Remarks:
The Test item was applied neat to the tissues
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
Acceptability of the Quality Control Data of the Skin Model with Reference to Historical Batch Data:
The negative control OD values were 1.802, 1.909 and 1.903 and, thus, in the range of ≥0.8 and ≤3.0.

Acceptability of the Positive and Negative Control Data:
After treatment with the negative control (DPBS-buffer) the mean OD was 1.871 (standard deviation: 3.91%) and, thus, higher than the historically established threshold of 1.423.
After treatment with the positive control (5% aqueous solution of sodium dodecyl sulfate) the mean viability value was 1.33% (standard deviation: 6.71%) and, thus, lower than the historically established threshold of 3.21%.

Variability of the Data:
The standard deviation between the three tissues replicates treated with the test item was 6.18% and, thus, ≤18%. The standard deviations between the three tissue replicates of the negative control and the positive control were 3.19% and 6.71%, respectively, and, thus, ≤18%.

The study met all acceptance criteria.

Any other information on results incl. tables

 Group Time / [min]  Mean OD  Mean Relative viability [%]
 Negative Control 42  1.871 100.00 
 Positive Control 42

0.025

1.33

 Test Material

42

1.824

97.48

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
UN GHS: No Category (according to OECD TG 439)
Conclusions:
This study was performed according to GLP and the methods applied are fully compliant with OECD TG 439. Under the conditions of the present study, the test item Art. 201386 (B-2O-O5) is not considered to possess an irritant potential to skin (UN GHS: No Category).
Executive summary:

This study was performed according to GLP and the methods applied are fully compliant with OECD TG 439. Under the conditions of the present study, the test item Art. 201386 (B-2O-O5) is not considered to possess an irritant potential to skin (UN GHS: No Category).