Registration Dossier

Toxicological information

Skin irritation / corrosion

Currently viewing:

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
December 20, 2019 - May 28, 2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2020
Report date:
2020

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
yes
Remarks:
Due to the volatility of the test substance, the incubation period was performed under a safety hood. Additional test substance also had to be sent to the testing laboratory.
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
(Z)-non-6-enal
EC Number:
218-900-9
EC Name:
(Z)-non-6-enal
Cas Number:
2277-19-2
Molecular formula:
C9H16O
IUPAC Name:
(Z)-non-6-enal
Test material form:
liquid

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Source strain:
other: 00267
Justification for test system used:
The test system is designed to mimic human skin, the target organ of interest.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Normal human epidermal keratinocytes
- Tissue batch number(s): 30858
- Date of initiation of testing: Jan. 9, 2020

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 degrees C
- Temperature of post-treatment incubation (if applicable): 37 degrees C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: 15 times with enough DPBS to fill the well, then submerged 3 times with 150 mL of DPBS
- Observable damage in the tissue due to washing: No
- Modifications to validated SOP:

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration:5 mg/mL in PBS
- Incubation time: 3 hour
- Wavelength: 570 nm
- Filter bandwidth: +/- 30 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: 1.598
- Barrier function: 6.45 hr
- Contamination: Sterile

NUMBER OF REPLICATE TISSUES: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- N. of replicates: 3

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 3

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if the viability is less than 50%.
- The test substance is considered to be non-corrosive to skin if the viability is greater than or equal to 50%.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 uL
- Concentration (if solution): 100%

VEHICLE
- Amount(s) applied (volume or weight with unit): None

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 uL
- Concentration (if solution): 100% DPBS

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 uL
- Concentration (if solution): 5% SDS
Duration of treatment / exposure:
25 minutes under a safety hood, then 35 minutes on a warming plate in a safety hood.
Duration of post-treatment incubation (if applicable):
24 hrs, then added to new wells containing 0.9 mL incubation medium and post-incubated for an additional 18 hrs.
Number of replicates:
3

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Average
Value:
3.7
Negative controls validity:
valid
Positive controls validity:
valid

Any other information on results incl. tables

 

Relative Tissue Viability (%)

Average Viability (%)

Standard Deviation (%)

Negative Control 1

99.6

100

7.1

Negative Control 2

93.1

Negative Control 3

107.3

Positive Control 1

3.6

3.7

0.1

Positive Control 2

3.8

Positive Control 3

3.7

Test Item 1

3.6

3.7

0.3

Test Item 2

3.4

Test Item 3

4.0

 

Applicant's summary and conclusion

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
The tissue viability was less than 50% at one hour of treatment. The test substance is therefore considered to be an irritant to skin.
Executive summary:

The potential for skin irritancy was studied in an OECD Guideline 439 in vitro human skin model test.  Model human epidermis was exposed to the test substance for one hour.  Due to volatility of the test substance, the incubation period was done under a safety hood.  Three replicates were performed.  The average percent viability for the test substance was 3.7%.  The test substance is therefore considered to be a category 2 skin irritant.