Registration Dossier

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
19/04/2016-22/04/2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2016

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Version / remarks:
Version 02A
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
Aspect : liquid
Color : yellow
Storage conditions : room temperature
Test item nature : cosmetic ingredient
Expiry date : 09/02/2017
Physical state at 20°C : Yes
Purity : NA (mixture)
Homogeneity : yes
pH : 3.8

In vitro test system

Test system:
human skin model
Remarks:
SkinEthic(TM) RHE model, 0.5 cm²
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
foreskin from a single donor
Source strain:
other: PK2 PKP.ABC-4
Details on animal used as source of test system:
Batch n° 16-RHE-040
Storage : Prepared/packaged using aseptic techniques. Stored in an incubator at 37°C, 5% CO2 with saturated humidity

Vehicle:
not specified
Details on test system:
Biological safety (blood of the donor) :
- absence of H1V1 and 2 antibodies
- absence of hepatitis C antibodies
- absence of hepatitis B antigen HBs
Biological safety (epidermal of the donor)
- absence of mycoplasma

Quality controls :
- Histological observation : 6.5 cell layers / absence of significant histological abnormalities
- Cell viability : O.D. = 1.1 (CV = 4.2%)
- Barrier function integrity test : 8.1h

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

REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps:
- Observable damage in the tissue due to washing:
- Modifications to validated SOP:
DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Spectrophotometer: yes
- Wavelength: 540 nm

NUMBER OF INDEPENDENT TESTING RUNS / EXPERIMENTS TO DERIVE FINAL PREDICTION:
PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if [complete, e.g. if the mean TER value is less than or equal to 5 kΩ and the skin disk is obviously damaged, or if the mean TER value is less than or equal to 5 kΩ, and the skin disc is showing no obvious damage, but the mean disc dye content is greater than or equal to the mean disc dye content of the 10M HCl positive control obtained concurrently.]
- The test substance is considered to be non-corrosive to skin if [complete, e.g. if the mean TER value obtained for the test substance is greater than 5 kΩ, or if the mean TER value is less than or equal to 5 kΩ, and the skin disc is showing no obvious damage, and the mean disc dye content is well below the mean disc dye content of the 10M HCl positive control obtained concurrently.]
- Justification for the selection of the cut-off point(s) if different than recommended in TG 430:
Control samples:
yes, concurrent negative control
yes, concurrent no treatment
yes, concurrent vehicle
Amount/concentration applied:
TEST MATERIAL
- Amount : 16 µL +/- 0.5 µL (treatment or control)
Duration of treatment / exposure:
42 minutes
Duration of post-treatment incubation (if applicable):
42 hours
Number of replicates:
3 epidermises

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1
Value:
ca. 93.2
Vehicle controls validity:
not specified
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
2
Value:
ca. 106.9
Vehicle controls validity:
not specified
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3
Value:
ca. 101.9
Vehicle controls validity:
not specified
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
Test validation:
The positive control shows a viability percentage of 2,3% ( lower than 40%) and all criteria are fulfilled, this validates the test.
The negative control shows a viability percentage of 100%

Any other information on results incl. tables

The test item shows a viability percentage of 100,7% (mean value of the 3 experiments) and therefore is considered as non irritant.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Executive summary:

Under the retained experimental conditions and according to the CLP regulation, the test item must not be classified. No symbol, risk phrase, no signal word or hazard statement is required.