Registration Dossier

Administrative data

Description of key information

Substance tested in chicken egg membrane (in vitro), rabbit eyes (in vivo) and rabbit skin (in vivo) tests.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP and OECD/EC guidelines
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Qualifier:
according to
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
GLP compliance:
yes (incl. certificate)
Test material information:
Composition 1
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, D-97633 Sulzfeld
- Age at study initiation: 17 weeks
- Weight at study initiation: 3.3 - 3.9 kg
- Housing: ABS plastic rabbit cages, floor 4200 cm2
- Diet (e.g. ad libitum): Free access to autoclaved hay and to Altromin 2123 maintenance diet for rabbits
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 +/-3 degrees C
- Humidity (%): 55 +/- 10%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
water
Controls:
other: animals were treated on one side, the other side was used as untreated control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5g
Duration of treatment / exposure:
4 hours
Observation period:
72 hours
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: 6 cm2
- Type of wrap if used: gauze patch held in place with non-irritating tape and fixed with a semi-occlusive dressing
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 1 hour
Score:
0
Max. score:
4
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 24 hours
Score:
0
Max. score:
4
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 48 hours
Score:
0
Max. score:
4
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 72 hours
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 1 hour
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 24 hours
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 48 hours
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 72 hours
Score:
0
Max. score:
4
Irritant / corrosive response data:
No irritation effects were observed in any of the test animals
Other effects:
No significant clinical signs of toxicity were observed in any of the test animals and there were no significant bodyweight changes.
Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The substance is not irritating to rabbit skin.
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:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to OECD guidelines and GLP.
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF, 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes
Test material information:
Composition 1
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River France, L’Arbresle Cedex, France
- Age at study initiation: Between 12 and 24 weeks old
- Weight at study initiation: At least 1.5 kg
- Housing: Individually housed in labelled cages with performated floors and shelters.
- Diet (e.g. ad libitum): Pelleted diet for rabbits at approximately 100g per day. Hay and wooden sticks were also available.
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: At least 5 days before the start of treatment, under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 25°C
- Humidity (%): 40 to 70%
- Air changes (per hr): At least 10 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hour light, 12 hour dark cycle

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Instillation on average of 82.9 mg (range 82.3 to 83.2 mg) of the test substance (a volume of approximately 0.1 mL).


Duration of treatment / exposure:
No test substance was observed on the eye at the first one hour observation check. This indicates it would have been removed by physiological mechanisms. Remnants of the test substance were noticed on the outside of the eyelid in one animal on Day 1.

If the test substance wasn't removed from the eye by physiological mechanisms at the first observation period of approximately 1 hour, the eye was rinsed with saline or distilled water.
Observation period (in vivo):
14 day observation period in surviving animals.
Number of animals or in vitro replicates:
3 (Study conducted in a stepwise manner and was started by treatment of a single rabbit (sentinel)). The two remaining animals were treated in a similar manner up to 3 weeks later, after considering the degree of eye irritation observed in the first animal.
Details on study design:
Eye irritation was assessed using the standard approach in the OECD 405 guidelines, which looks at corneal irritation (including the area of the cornea involved), Iris irritation, conjunctival irritation, chemosis, and evidence of discharge.

Where standard lighting was considered inadequate for observing minor effects, eye examinations were performed using an ophthalmic examination lamp.

Green staining after fluorescein treatment was used to determine the % of total corneal area effects.
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
of animals 1 and 2.
Time point:
other: 24, 48 and 72 hours
Score:
0.3
Max. score:
0.3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
iris score
Basis:
mean
Remarks:
of animals 1 and 2
Time point:
other: 24, 48 and 72 hours
Score:
0.15
Max. score:
0.3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
of animals 1 and 2
Time point:
other: 24, 48 and 72 hours
Score:
2.15
Max. score:
2.3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Remarks:
chemosis
Basis:
mean
Remarks:
of animals 1 and 2.
Time point:
other: 24, 48 and 72 hours
Score:
1.65
Max. score:
2
Reversibility:
fully reversible within: 14 days
Irritant / corrosive response data:
Two animals showed similar eye irritation effects in the cornea, iris and conjunctivae approximately 1 hour after instillation.
Cornea: The corneal injury consisted of opacity and epithelial damage and resolved within 7 days.
Iris: Iridial irritation was observed and resolved within 24 hours in one of the animals, and within 48 hours in the other.
Conjunctivae: The irritation of the conjunctivae consisted of redness, chemosis, and discharge and completely resolved within 14 days in both animals.

There was no evidence of ocular corrosion.
Other effects:
One animal was found undergoing severe spasms followed by a comatose appearance approximately 45 minutes after instillation of the test substance. After this, the animal was sacrificed for humane reasons. Post mortem examination showed eye effects similar to the surviving rabbits.

As this effect was not seen in the two remaining rabbits (which both showed similar eye irritation effects and full reversibility) and that the sacrificed animal had mostly slightly lesser eye irritation findings, this effect is not considered to be evidence of systemic toxicity and was most likely a stress response of a potentially (prior to treatment) compromised animal.

Mean value eye irritations scores:

Animal

Mean 24, 48 and 72 hours

Corneal Opacity

Iris

Conjunctivae

Redness

Chemosis

1

0.3

0.3

2.0

1.3

2

0.3

0.0

2.3

2.0

3

-

-

-

-

Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test substance showed eye irritation effects in two animals which were fully reversible by Day 14 of the observation period. One animal that was sacrificed for humane reasons had effects not attributed directly to treatment and systemic toxicity of the test substance.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The test substance was found to be not irritating to rabbit skin in an in vivo skin irritation/corrosion test.

In an in vivo eye irritation/corrosion test, the test substance was found to show irritation shortly after test substance instillation. The animals had shown complete recovery by day 14 of observation.


Justification for selection of skin irritation / corrosion endpoint:
In vivo study conducted to OECD guidance and GLP

Justification for selection of eye irritation endpoint:
In vivo study conducted to OECD guidance and GLP

Effects on eye irritation: irritating

Justification for classification or non-classification

As the test substance showed no evidence of skin irritation or corrosion in an in vivo skin irritation/corrosion test, no classification for skin irritation/corrosion effects are warranted for this substance.

With regards to eye irritation, as the mean score following grading at 24, 48 and 72 hours for Conjunctival redness was >/= 2 (2.15) in 2 animals which showed recovery within the observation period. One animal was observed to have spasms shortly after instillation of the test material and was killed for human reasons. This is not thought to be attributed to treatment and most likely a stress reaction in a compromised rabbit and as such it is considered that the test material does not cause irreversible effects on the eye.

Due to the mean score for conjunctival redness, the classification as per the CLP regulation (EC No. 1272/2008, as amended) as Irritating to eyes (category 2) is considered appropriate for this substance