Castor oil, oxidized

Endpoint summary

Description of key information

Skin irritation: not irritating (in vitro, EU method B.46, based on RA substances Blown Linseed oil and Blown rapeseed oil)
Eye irritation: not irritating (in vivo, OECD 405, based on RA substance Blown Linseed oil)
Respiratory irritation: no information available, but not expected based on absence of skin and eye irritating properties

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation / corrosion

Remarks:

other: in vitro

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

23-feb-2010 to 01-mar-2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

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.

Qualifier:

according to guideline

Guideline:

other: OECD Draft Proposal for a New Guideline (In Vitro Skin Irritation: Reconstructed Human Epidermis (RhE) Test Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EU method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 µl of the undiluted test substance

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.

Irritation parameter:

other: percentage viability

Basis:

other: percentage of control

Time point:

other: 15 minutes

Score:

109

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

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 linseed oil compared to the negative control tissues was 109%. Since the mean relative tissue viability for Blown linseed oil was above 50% after 15 minutes treatment Blown linseed oil is considered to be non-irritant.

Finally, it is concluded that this test is valid and that Blown linseed 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

Reference

Endpoint:

eye irritation: in vivo

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

06 April 2010 - 22 April 2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

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.

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.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Not irritating

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

For both BLO and BRO, two in vitro studies are available that have been included as key studies: a skin corrosion test according to OECD 431(In vitro skin corrosion: Human Skin Model Test) and skin irritation test according to EU method B.46 (In vitro skin Irritation: Reconstructed Human Epidermis Model Test)/OECD Draft Proposal for a New Guideline: In Vitro Skin Irritation: Reconstructed Human Epidermis (RhE) Test Method). The tested BLO had a viscosity of 1 Poise in both studies, while BRO had a viscosity of 18 Poise. In vivo studies are not available, and are not required as the in vitro study has recently been accepted as stand-alone study for this endpoint.

In the skin corrosion tests, 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 BLO compared to the negative control tissues was 97% and 102%, respectively, while for BRO this was 97%. Because the mean relative tissue viability for BLO and BRO were not below 50% after the 3-minute treatment and not below 15% after the 1-hour treatment BLO and BRO are considered to be not corrosive.

In the skin irritation tests, skin irritation is expressed as the remaining cell viability after exposure to the test substance (EPISKIN Standard ModelTM). The relative mean tissue viability obtained after 15 minutes treatment with BLO and BRO compared to the negative control tissues was 109% and 106%, respectively. Since the mean relative tissue viability for both oils was above 50% after 15 minutes treatment BLO and BRO are considered to be non-irritant.

It was concluded that BLO and BRO are not irritating and not corrosive to the skin. The result of are read across to BCO. This is considered justified because 1) of the reasons mentioned above (Human toxicity) and because 2) two oils in this category showed to be not irritating and not corrosive to the skin.

 

Eye irritation

Two in vitro key studies are available and one in vivo study. An in vitro study according to OECD 437 and one in vivo study according to OECD 405 are conducted with BLO, while a second in vitro study according to OECD 437 is available for BRO. BLO had a viscosity of 1 Poise, while BRO had a viscosity of 18 Poise. In the in vitro study with BLO, a Bovine Corneal Opacity and Permeability test (BCOP) according to OECD 437, it was shown that that BLO did not induce ocular irritation, resulting in a mean in vitro irritancy score of 0 after 240 minutes of treatment and an irritancy score of 0.3 after 10 minutes of treatment, respectively. Similar results were obtained in the in vitro study with BRO, which was also a BCOP test in accordance with OECD 437. BRO did not induce ocular irritation, resulting in an irritancy score of 0.3 after 10 minutes of treatment. In the in vivo study with rabbits according to OECD 405, BLO did not induce any effect after 72 h. It was concluded that BLO and BRO are not irritating to the eye.

For BCO no data are available, therefore read across was conducted. The in vitro results of BLO and BRO are read across to BCO. This is considered justified because 1) of the reasons mentioned above (Human toxicity) and because 2) the in vitro results for 2 oils in this category showed that these oils were not irritating to the eye and 3) the in vivo study with 1 oil confirmed that these oils are not irritating to the eye.

Justification for selection of skin irritation / corrosion endpoint:
The selected study is one of the four available in vitro key studies with read across substances Blown linseed oil and Blown rapeseed oil.

Justification for selection of eye irritation endpoint:
The selected study is the only available in vivo study for this endpoint with read across substance Blown linseed oil.

Justification for classification or non-classification

Based on the available information, Blown castor oil does not have to be classified for skin, eye, or respiratory irritation in accordance with the classification criteria outlined in Annex VI of 67/548/EEC and Annex I of 1272/2008.