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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
The study has been performed according to OECD and/or EC guidelines and according to GLP principles. A reliability of 2 is assigned in accordance with the ECHA Practical guide #6 on the reporting of read-across in IUCLID, due to the read-across purpose.
Cross-reference
Reason / purpose:
reference to same study
Reference
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
See attached justification
Reason / purpose:
read-across source
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
15 minutes
Value:
123
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other:
Remarks:
viability: percentage of control. Time point: 15 minutes.
Other effects / acceptance of results:
The positive control had a mean cell viability after 15 minutes exposure of 22%. The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 18%, indicating that the test system functioned properly.

Interpretation of results:
other: Not irritating
Remarks:
Based on CLP criteria
Conclusions:
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 Standolized linseed oil compared to the negative control tissues was 123%. Since the mean relative tissue viability for Standolized linseed oil was above 50% after 15 minutes treatment Standolized linseed oil is considered to be non-irritant.

The positive control had a mean cell viability after 15 minutes exposure of 22%. The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 18%, indicating that the test system functioned properly.

Finally, it is concluded that this test is valid and that Standolized linseed oil is non-irritant in the in vitro skin irritation test under the experimental conditions described in this report.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Version / remarks:
OECD Draft Proposal for a New Guideline: In Vitro Skin Irritation: Reconstructed Human Epidermis (RhE) Test Method
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Standolized linseed oil
- Substance type: Hazy yellow/brown viscous liquid
- Physical state: liquid
- Stability under test conditions: Stable
- Storage condition of test material: At room temperature in the dark

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
other: human-derived epidermal keratinocytes. (highly differentiated and stratified epidermis model comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum)
Cell source:
other: not specified
Source strain:
not specified
Details on animal used as source of test system:
SOURCE ANIMAL
- Source: adult human-derived epidermal keratinocytes
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN Standard ModelTM (EPISKIN-SMTM, 0.38 cm2) This model is a three-dimensional human epidermis model, which consists of adult human-derived epidermal keratinocytes which have been seeded on a dermal substitute consisting of a collagen type I matrix coated with type IV collagen. The keratinocytes were cultured for 13 days, which results in a highly differentiated and stratified epidermis model comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum.
- Tissue batch number: Lot no.: 10-EKIN-006

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: Washing with phosphate buffered saline 15 minutes after exposure.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
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.

PREDICTION MODEL / DECISION CRITERIA
If the mean relative tissue viability was above 50% after 15 minutes treatment the substance is considered to be non-irritant.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 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
Duration of post-treatment incubation (if applicable):
Post incubation period: 42 hours

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
15 minutes
Value:
123
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other:
Remarks:
viability: percentage of control. Time point: 15 minutes.
Other effects / acceptance of results:
The positive control had a mean cell viability after 15 minutes exposure of 22%. The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 18%, indicating that the test system functioned properly.

Applicant's summary and conclusion

Interpretation of results:
other: Not irritating
Remarks:
Based on CLP criteria
Conclusions:
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 Standolized linseed oil compared to the negative control tissues was 123%. Since the mean relative tissue viability for Standolized linseed oil was above 50% after 15 minutes treatment Standolized linseed oil is considered to be non-irritant.

The positive control had a mean cell viability after 15 minutes exposure of 22%. The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 18%, indicating that the test system functioned properly.

Finally, it is concluded that this test is valid and that Standolized linseed oil is non-irritant in the in vitro skin irritation test under the experimental conditions described in this report.