Registration Dossier

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch identification ZH 291 A1 Fr.2-4

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature



In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm TM, EPI-200, MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia
- Tissue batch number(s): Lot Number 23362
- Date of initiation of testing: 2016-09-06

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature (25 min, laminar flow hood) followed by 37 °C (35 min, incubator)
- Temperature of post-treatment incubation: 37 °C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps: at least one washing step with sterile PBS
- Observable damage in the tissue due to washing: No

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL
- Incubation time: 3 h
- Spectrophotometer: Sunrise Absorbance Reader
- Wavelength: 570 nm

NUMBER OF REPLICATE TISSUES:
3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Killed tissues
- Procedure used to prepare the killed tissues: freezing
- N. of replicates : 3

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be irritating to skin if the mean relative tissue viability with a test material is less than or equal to 50%.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: 30 µL

NEGATIVE CONTROL
- Amount applied: 30 µL PBS, sterile

POSITIVE CONTROL
- Amount applied: 30 µL sodium dodecyl sulfate in water
- Concentration: 5 % (w/v)
Duration of treatment / exposure:
25 min (laminar flow hood) followed by 35 min (incubator)
Duration of post-treatment incubation (if applicable):
42 ± 4 h
Number of replicates:
3

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Value:
73.2
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation

Any other information on results incl. tables

 

 

 

Tissue 1

Tissue 2

Tissue 3

Mean

SD

CV [%]

Test substance

 

Mean OD570

1.248

1.377

1.375

1.333

 

 

Viability [% of NC]

68.5

75.6

75.5

73.2

4.1

5.6

NC

 

Mean OD570

1.973

1.838

1.654

1.821

 

 

Viability [% of NC]

108.3

100.9

90.8

100.0

8.8

8.8

PC

 

Mean OD570

0.054

0.063

0.065

0.060

 

 

Viability [% of NC]

3.0

3.4

3.5

3.3

0.3

9.2

 

The negative results derived with the Skin Irritation Test on pyranyl acetate pure alone were sufficient for a final assessment. Therefore further testing in the Skin Corrosion Test was waived.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met