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Description of key information

Not irritating (OECD 431, In Vitro Skin Corrosion: Human Skin Model Test), not irritating (OECD Draft Guideline, In Vitro Skin Irritation), not irritating (OECD 437, BCOP test method) and not irritating (OECD 405, Acute Eye Irritation / Corrosion)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
02-Mar-2010 to 05-Mar-2010
Reliability:
1 (reliable without restriction)
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.
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test system:
human skin model
Source species:
human
Cell type:
other: The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis.
Cell source:
other: not specified
Source strain:
other: not applicable
Justification for test system used:
According to OECD 431
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm Skin Model (EPI-200,no.: 12935 kit H).
The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those foundin vivo. The EpiDerm tissues (surface 0.6 cm²) were cultured on polycarbonate membranes of 10 mm cell culture inserts.

TEST SITE
- % coverage: 0.6 cm2

REMOVAL OF TEST MATERIAL AND CONTROLS
- Washing: physiological saline
- Time after start of exposure: 3 minutes and 1 hour

SCORING SYSTEM:
- Percentage viability

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be corrosive to skin if [complete, e.g. the viability after 3 minutes exposure is less than 50%, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15%.]
- The test substance is considered to be non-corrosive to skin if [complete, e.g. the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is greater than or equal to 15%.]
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 µl of the undiluted test substance

NEGATIVE CONTROL
- Amount(s) applied (volume or weight with unit): 50 µl water

POSITIVE CONTROL
- Amount(s) applied (volume or weight with unit): 50 µl KOH
- Concentration (if solution): 8N
Duration of treatment / exposure:
- 3 minutes and 1 hour exposure times
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3 minutes
Value:
97
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1 hour
Value:
97
Negative controls validity:
valid
Positive controls validity:
valid
Interpretation of results:
other:
Remarks:
not corrosive according to CLP criteria
Conclusions:
The positive and negative controls were within the historical control data. Skin corrosion is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 3-minute and 1-hour treatments with Blown rapeseed oil compared to the negative control tissues was 97%. Because the mean relative tissue viability for Blown rapeseed oil was not below 50% after the 3-minute treatment and not below 15% after the 1-hour treatment Blown rapeseed oil is considered to be not corrosive.

Finally, it is concluded that this test is valid and that Blown rapeseed oil is not corrosive in the in vitro skin corrosion test under the experimental conditions described in the report.
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09-Mar-2010 to 11-Mar-2010
Reliability:
1 (reliable without restriction)
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.
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 system:
human skin model
Source species:
human
Cell type:
other: adult human-derived epidermal keratinocytes
Cell source:
other: not specified
Source strain:
other: not applicable
Justification for test system used:
According to OECD 439
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
EPISKIN Standard ModelTM(EPISKIN-SMTM, 0.38 cm2, Lot no.: 10-EKIN-008).
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.

TEST SITE
- % coverage: 0.38 cm2

REMOVAL OF TEST MATERIAL AND CONTROLS
- Washing: 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.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied: 10 µl

NEGATIVE CONTOL:
- Amount(s) applied: 10 µl Phosphate buffered saline

POSITIVE CONTROL
- Amount(s) applied: 10 µl
- Concentration: 5% (aq) Sodium dodecyl sulphate
Duration of treatment / exposure:
Exposure:15 minutes
Duration of post-treatment incubation (if applicable):
Post incubation period: 42 hours
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
15 minutes
Value:
106
Negative controls validity:
valid
Positive controls validity:
valid
Interpretation of results:
other:
Remarks:
not irritating, CLP criteria not met.
Conclusions:
The positive and negative controls were within the historical control data. 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 Blown rapeseed oil compared to the negative control tissues was 106%. Since the mean relative tissue viability for Blown rapeseed oil was above 50% after 15 minutes treatment Blown rapeseed oil is considered to be non-irritant.

Finally, it is concluded that this test is valid and that Blown rapeseed oil is non-irritant in the in vitro skin irritation test under the experimental conditions described in the report.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
06 April 2010 - 22 April 2010
Reliability:
1 (reliable without restriction)
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.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Harlan, Belton, Leics, England
- Age at study initiation: Animals used within the study were at least 6 weeks old
- Weight at study initiation: body weights were at least 1.0 kg.
- Housing: labeled cages with perforated floors (Ebeco, Germany, dimensions 67 x 62 x 55 cm) and shelters (Ebeco, Germany, dimensions 40 x 32 x 23 cm).
- Diet (e.g. ad libitum): Pelleted diet for rabbits (Global Diet 2030 from Harlan Teklad®, Mucedola, Milanese, Italy) approximately 100 grams per day. Hay (TecniLab-BMI BV, Someren, The Netherlands) was provided at least three times a week.
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: Acclimatization period was at least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.6 – 19.4ºC
- Humidity (%): 44 - 73%
Cleaning procedures in the room might have caused the temporary fluctuations above the optimal maximum level of 70% for relative humidity. Based on laboratory historical data, these fluctuations were considered not to have affected the study integrity.
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day

IN LIFE DATES: From: 06 April 2010 To: 22 April 2010
Vehicle:
unchanged (no vehicle)
Controls:
other: One eye of each animal remained untreated and served as the reference control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
Duration of treatment / exposure:
Single instillation on Day 1
Observation period (in vivo):
The eyes of each animal were examined approximately 1, 24, 48 and 72 hours after instillation of the test substance.
Number of animals or in vitro replicates:
3 Males
Details on study design:
STUDY DESIGN
The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 13 days later, after considering the degree of eye irritation observed in the first animal.

TREATMENT
Each animal was treated by instillation of 0.1 mL of the test substance in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control.

Immediately after the 24-hour observation, a solution of 2% fluorescein (Merck, Darmstadt, Germany) in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.

After the final observation, the animals were sacrificed by intra-venous injection of Euthasol® 20% (AST Farma BV, Oudewater, The Netherlands).


REMOVAL OF TEST SUBSTANCE
- Washing (if done): No

OBSERVATION
The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

SCORING SYSTEM:
CORNEAL IRRITATION
Opacity: degree of density (area most dense taken for reading)
0: No ulceration or opacity (may include slight dulling of normal luster)
1: Scattered or diffuse areas of opacity, details of iris clearly visible
2: Easily discernible translucent area, details of iris slightly obscured
3: Nacreous area, no details of iris visible, size of pupil barely discernible
4: Opaque cornea, iris not discernible through the opacity

Area of cornea involved:
0: No ulceration or opacity
1: One quarter or less but not zero
2: Greater than one quarter, but less than half
3: Greater than half, but less than three quarters
4: Greater than three quarters, up to whole area

IRIS
0: Normal
1: Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia,
or injection, any of these or combination thereof, iris still reacting to light
(sluggish reaction is positive)
2: No reaction to light, hemorrhage, gross destruction (any or all of these)

CONJUNCTIVAL IRRITATION
Redness (refers to palpebrae and sclera, excluding cornea and iris):
0: Blood vessels normal
1: Some blood vessels definitely hyperaemic (injected)
2: Diffuse, crimson color, individual vessels not easily discernible
3: Diffuse beefy red

Chemosis (refers to lids and/or nictitating membranes):
0: No swelling
1: Any swelling above normal (includes nictitating membranes)
2: Obvious swelling with partial eversion of lids
3: Swelling with lids about half closed
4: Swelling with lids more than half closed

Discharge:
0: No discharge (may include small amounts observed in inner canthus of normal animals)
1: Any amount different from normal and/or lacrimation
2: Discharge with moistening of the lids and hairs just adjacent to lids
3: Discharge with moistening of the lids and hairs (considerable area around the eye)

Where standard lighting was considered inadequate for observing minor effects, eye examinations were performed using an ophthalmic examination lamp.
In cases of equivocal results when comparing the treated and untreated eyes, the illustrated guide from the Consumer Product Safety Commission, Washington, D.C. 20207 was used for additional control purposes.
Irritation parameter:
cornea opacity score
Remarks:
Opacity
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
0
Max. score:
4
Reversibility:
other: No findings noted
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
0
Max. score:
2
Reversibility:
other: No findings.noted.
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
0.11
Max. score:
3
Reversibility:
fully reversible within: for 2 animals in 24 hours and for 1 animal in 48 hours
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 24 hours (=1 animal)
Irritant / corrosive response data:
Instillation of 0.1 mL of Blown linseed oil into one eye of each of three rabbits resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation completely resolved within 24 hours in two animals and within 48 hours in the other animal.

No iridial irritation or corneal opacity were observed, and treatment of the eyes with 2% fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage.

See also attached tables.
Other effects:
Coloration / Remnants
Remnants of the test substance were present on the outside of the eyelids on Days 1 and 2.

Toxicity / Mortality
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

For all details on the properties of the source material, the description of the analogue approach justification and the data matrix, see the attached document

Interpretation of results:
other:
Remarks:
not irritating, CLP criteria not met.
Conclusions:
The results obtained with the substance Blown linseed oil are suitable for the regulatory purpose of REACH for the substance Blown rapeseed oil: Not irritating
Endpoint:
eye irritation: in 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 for cross-reference:
read-across source
Irritation parameter:
cornea opacity score
Remarks:
Opacity
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
0
Max. score:
4
Reversibility:
other: No findings noted
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
0
Max. score:
2
Reversibility:
other: No findings.noted.
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
0.11
Max. score:
3
Reversibility:
fully reversible within: for 2 animals in 24 hours and for 1 animal in 48 hours
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 24 hours (=1 animal)
Irritant / corrosive response data:
Instillation of 0.1 mL of Blown linseed oil into one eye of each of three rabbits resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation completely resolved within 24 hours in two animals and within 48 hours in the other animal.

No iridial irritation or corneal opacity were observed, and treatment of the eyes with 2% fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage.

See also attached tables.
Other effects:
Coloration / Remnants
Remnants of the test substance were present on the outside of the eyelids on Days 1 and 2.

Toxicity / Mortality
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

For all details on the properties of the source material, the description of the analogue approach justification and the data matrix, see the attached document

Interpretation of results:
other:
Remarks:
not irritating, CLP criteria not met.
Conclusions:
The results obtained with the substance Blown linseed oil are suitable for the regulatory purpose of REACH for the substance Blown rapeseed oil: Not irritating
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18-Mar-2010
Reliability:
1 (reliable without restriction)
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.
Qualifier:
according to guideline
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Bovine eyes from young cattle were obtained from the slaughterhouse, where the eyes were excised by a slaughterhouse employee as soon as possible after slaughter
Vehicle:
not specified
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
- Amount(s) applied: 750 µl per cornea

NEGATIVE CONTROL
- Amount(s) applied: 750 µl of physiological saline per cornea

POSITIVE CONTROL
Amount(s) applied: 750 µl per cornea
Concentration: 10% (w/v) Benzalkonium Chloride
Duration of treatment / exposure:
- Exposure: 10 minutes


Duration of post- treatment incubation (in vitro):
- Post incubation period: 120 minutes
Details on study design:
TEST SITE
- Isolated bovine cornea

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 10 minutes

SCORING SYSTEM:
- After the 10 minutes exposure and washing step, the corneas were incubated for 120 +/- 10 minutes at 32 °C. After the completion of the incubation period opacity determination was performed.
- The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:

In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).

TOOL USED TO ASSESS SCORE:
- opacitymeter and microplate reader

DATA EVALUATION:
A test substance that induces an IVIS ≥ 55.1 is defined as a corrosive or severe irritant
Irritation parameter:
in vitro irritation score
Run / experiment:
10 minutes
Value:
0.3
Negative controls validity:
valid
Positive controls validity:
valid
Interpretation of results:
other:
Remarks:
not severe irritant or corrosive, CLP criteria not met.
Conclusions:
The positive and negative controls were within the historical control data.Blown rapeseed oil did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of 0.3 after 10 minutes of treatment. Finally, it is concluded that this test is valid and that Blown rapeseed oil is a non irritant in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in the report.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The acute eye irritation (OECD 405) is read-across from a supporting substance (structural analogue or surrogate).

Justification for classification or non-classification

Based on the results obtained in the 4 studies investigating skin and eye irritation (in vitro and in vivo), it is shown that the substance does not need to be classified.