Registration Dossier

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21-Jul-2014 to 28-Jul-2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report date:
2014

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics
IUPAC Name:
Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics
- Substance type: Clear colourless liquid
- Physical state: Liquid
- Storage condition of test material: At room temperature in the dark

Test system

Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 µl

NEGATIVE CONTOL:
- Amount(s) applied (volume or weight with unit): 10 µl Phosphate buffered saline

POSITIVE CONTROL
- Amount(s) applied (volume or weight with unit): 10 µl
- Concentration (if solution): 5% (aq) Sodium dodecyl sulphate
Duration of treatment / exposure:
Exposure:15 minutes
Post incubation period: 42 hours
Details on study design:
TEST SITE
- Area of exposure: human epidermis model
- % coverage: 0.38 cm2

REMOVAL OF TEST SUBSTANCE
- Washing (if done): phosphate buffered saline
- Time after start of exposure: 15 minutes

POST INCUBATION PERIOD
- 42 hours

SCORING SYSTEM:
- After a 42 hour incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment. Cell viability was calculated for each tissue as a percentage of the mean of the negative control tissues.

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
15-minute exposure
Value:
94
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
The absolute mean of the negative control was slightly below the historical control data range. It was concluded that the deviation had no effect on the study results.
Positive controls validity:
valid
Remarks:
A clear toxic effect was observed with the positive control. The low absolute mean OD570 value of the positive control, which was in the lower range of the historical control data range, resulted in a good measurement window.
Remarks on result:
no indication of irritation

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
An in vitro skin irritation study was also conducted with Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics (GS270) in accordance with OECD Test Guideline 439 and in compliance with GLP. The in vitro assay did not indicate any skin irritation potential.
Executive summary:

Skin irritation is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 15 minutes treatment with Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics compared to the negative control tissues was 94%. Since the mean relative tissue viability for the test substance was above 50% after 15 minutes treatment the test substance is considered to be non-irritant.

 

The positive control had a mean cell viability of 21% after 15 minutes exposure. The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was slightly below laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 7%, indicating that the test system functioned properly.