Registration Dossier

Administrative data

Description of key information

The test substance was found to be non-irritating to skin, while it was slightly irritating to eyes.

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
Study period:
From May 03, 2006 to May 06, 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Ferenc Sandor breeder, 2173 Kartal, Császár út 135, Hungary
- Animal health: Only animals in acceptable health condition were used for the test.
- Age of animals at arrival: 11 weeks old, adult albino rabbit
- Body weight range at treatment: 2639-3118 g (i.e., at the beginning of the study); 2720-3240 g (i.e., at the end of the study)
- Housing: Animals were housed individually in metal cages
- Diet: PURINA Base – Lap gr. diet for rabbit, ad libitum
- Water: Tap water from self-service watering system, ad libitum
- Acclimation period: Male: 8 d
-Animal identification: The individual identification was performed by numbered ear tag. The cages were marked with individual identity cards with information about study code, sex, dose group, cage number and individual animal number

ENVIRONMENTAL CONDITIONS
- Temperature: 20±3°C
- Humidity: 30-70%
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From: To: May 03, 2006 to May 06, 2006
Type of coverage:
occlusive
Preparation of test site:
clipped
Vehicle:
water
Remarks:
moistened with water to form a paste
Controls:
yes, concurrent no treatment
Amount / concentration applied:
500 mg (i.e., 0.5 g)
Duration of treatment / exposure:
4 h
Observation period:
1, 24, 48 and 72 h after the patch removal
Number of animals:
3 males
Details on study design:
PREPARATION OF THE TEST SUBSTANCE
The test substance was used in pure state, in a single dose. 0.5 g of test substance was applied to the test site. The test substance was moistened sufficiently with water to ensure good contact with the skin. Untreated skin of each animal served as control.

TEST PROCEDURE

Patch testing was used to detect primary irritating effects of the test substance. Three male animals in acceptable health condition were selected for this test. Each animal was examined 24 h before starting the test.

Approximately 24 h prior to the test the hair was clipped from the back and flanks of the animals.

The test substance was applied to an approximately 6 cm² area of intact skin. Sterile gauze pads (5x5 cm) were placed on the skin of rabbits. These gauze patches were kept in contact with the skin by a patch with a surrounding adhesive hypoallergenic plaster. The entire trunks of the animals were wrapped with plastic wrap for 4 h.

In the first step an initial test was performed using one animal. 1 h after application of the test substance to the sentinel animal, the application site was examined. Severe irritation or corrosive effect could not be found in the initial test, therefore the bandage was replaced and the exposure continued for a further 3 h (i.e., a total 4 h exposure). Two additional animals were involved for this study.

Duration of exposure: 4 h

REMOVAL OF TEST SUBSTANCE
After the treatment period the rest of the test substance was removed with body temperature water.

Scoring and Assessment of Local Reactions
The dermal irritation scores were evaluated according to the scoring system by Draize (1959). The animals were observed for two weeks, the duration of the study was sufficient to evaluate fully the reversibility or irreversibility of the effects observed.
Irritation parameter:
erythema score
Basis:
animal: 1, 2, and 3
Remarks:
(mean score)
Time point:
24/48/72 h
Score:
0.66
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks:
very slight erythema at 24 and 48 h
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal: 1, 2, and 3
Remarks:
(mean score)
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
1 h after the patch removal in all animals very slight erythema appeared on the treated skin surface.
24 and 48 h after the patch removal the observed signs did not change when compared to the previous observation.
72 h after the patch removal no primary irritation symptoms, like erythema and oedema or other signs occurred on the treated skin surfaces, so the study was terminated at this time.
During the study the control area was symptom-free.
Other effects:
General state and the behaviour of animals were normal throughout the study period.
There were no notable body weight changes during the contact and observation period.
Interpretation of results:
GHS criteria not met
Conclusions:
Under the test conditions, the test substance was considered to be slightly irritating to skin but all signs were fully reversible within 72 h. The substance is not classifiable according to CLP criteria.
Executive summary:

A study was performed to assess the skin irritation potential of the test substance in male New Zealand White rabbits according to OECD Guideline 404, EPA OPPTS 870.2500 and EU Method B.4, in compliance with GLP.

500 mg of test substance moistened with water was applied to an approximately 6 cm² area of intact skin. Sterile gauze pads (5x5 cm) were placed on the skin of rabbits surrounded by an adhesive hypoallergenic plaster. The entire trunks of the animals were wrapped with plastic wrap for 4 h. After the exposure period, all remnants of test substance were carefully removed from the skin with body temperature water. Animals were scored for irritation reactions after 60 minutes, and at 24, 48 and 72 h.

 

Except for very slight erythema at 24 h and 48 h, no other signs of irritation were observed in treated animals during the observation period. Also, the observed erythema was reversible with 72 h. Further, mean scores for dermal irritation for each animal were calculated to be 0.66 for erythema and 0.00 for edema formation. In addition, no clinical signs of systemic toxicity were observed.

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
Study period:
From May 04, 2006 to May 11, 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline 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
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Ferenc Sandor breeder, 2173 Kartal, Császár út 135, Hungary
- Animal health: Only animals in acceptable health condition were used for the test. Both eyes of each animal provisionally selected for testing were examined 24 h prior to the start of study. Animals showing eye irritation, ocular defects or pre-existing corneal injury were not used.
- Age of animals at arrival: 11 weeks old, adult albino rabbit
- Body weight range at treatment: 2614-3017 g (i.e., at the beginning of the study); 2739-3108 g (at the end of the study)
- Housing: Animals were housed individually in metal cages.
- Diet: PURINA Base – Lap gr. diet for rabbit, ad libitum
- Water: Tap water from self-supplying watering system, ad libitum
- Acclimation period: Male: 9 d
-Animal identification: The individual identification was performed by numbered ear tag. The cages were marked with individual identity cards with information about study code, sex, dose group, cage number and individual animal number

ENVIRONMENTAL CONDITIONS
- Temperature: 20±3°C
- Humidity: 30-70%
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From: To: May 04, 2006 to May 11, 2006
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
100 mg (i.e., 0.1 g)
Duration of treatment / exposure:
24 h
Observation period (in vivo):
1, 24, 48, 72 h and one week after the application of test substance
Number of animals or in vitro replicates:
3 males
Details on study design:
TEST PROCEDURE

Dosage
0.1 g of the test substance was used for the study in pure state, in a single dose. The absorption of the test substance was not determined.

Application of the Test Substance
Three male animals in acceptable health condition were selected for the test. Care was taken to select only those animals that had a normal eye condition, any with ocular lesions were rejected.

In the first step an initial test was performed using one animal. The test substance was instilled into the conjunctival sac of the left eye. The eyelids were held closed gently for several seconds to prevent the loss of the test substance. The contralateral eye served as control. Immediately after the administration of the test substance, an assessment of the initial pain reaction was made. After consideration of the ocular responses produced in the first animal, two additional animals were treated. Before the administration the treated eyes of animals were not anaesthetised, because the score of initial pain reaction was 0 in the first animal.

Duration of Exposure
The eyes of the test animals were not washed out after the application of test substance.

OBSERVATIONS AND SCORING

Clinical Observations

The eyes were examined at 1, 24, 48, 72 h then one week after the treatment. The duration of the observation period was sufficient for the statement of reversibility or irreversibility of changes. Any clinical signs of toxicity or signs of ill- health during the study were recorded. At the end of the observation period all animals were sacrificed by intramuscular injections of Ketamin and Xylazin followed by i. v. Euthanyl anaesthesia.

Scoring and Assessment of Local Reaction

The eye irritation scores were evaluated according to the scoring system by Draize (1959) and OECD 405 (24 April 2002).
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal: 1, 2, and 3
Remarks:
(mean score)
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 7 d
Remarks on result:
probability of mild irritation
Irritation parameter:
chemosis score
Basis:
animal: 1, 2, and 3
Remarks:
(mean score)
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Remarks:
(opacity)
Basis:
animal: 1 and 3
Remarks:
(mean score)
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 d
Remarks on result:
positive indication of irritation
Irritation parameter:
cornea opacity score
Remarks:
(opacity)
Basis:
animal #2
Remarks:
(mean score)
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal: 1, 2, and 3
Remarks:
(mean score)
Time point:
24/48/72 h
Score:
0
Max. score:
2
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
1h after treatment some hyperaemic blood vessels with obvious conjunctival swelling with partial eversion of the lids were found in every animal. In these animals the discharge moistened lids and hairs on considerable area around the treated eye. In all animals diffuse areas of opacity were observed on the cornea but the details of iris were clearly visible. Area of cornea involved was greater than three quarters, up to the whole area.

There were no effects observed in the iris alterations during the study period.

24 h after treatment in all animals some hyperaemic blood vessels were observed. Chemosis and increased discharge excretion were not found by this time. In two animals the degree and extension of cornea opacity did not change when compared to the previous observation. In another animal cornea opacity was not found by this time.

48 and 72 h after treatment in the animals the slight redness and degree and extension of cornea opacity did not change when compared to the previous observation.

One week after the treatment every animal was symptom-free.

One week after the treatment the study was terminated, since no primary irritation symptoms remained.
Other effects:
General state and the behaviour of animals were normal throughout the study period. There were no notable body weight changes during the contact and observation period.
Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
Under the test conditions, the test substance was considered to be slightly irritating to eyes. Due to the 24/48/72 h mean cornea opacity score of >=1 in 2/3 animals, a classification according to CLP criteria applies. The substance is considered to cause irritation in eyes.
Executive summary:

A study was conducted to assess the eye irritancy potential of the test substance in male New Zealand White rabbits according to OECD Guideline 405, EPA OPPTS 870.2400 and EU Method B.5, in compliance with GLP.

A single dose of 100 mg test substance was applied to the conjunctival sac of the left eye of each animal. The treated eyes were not washed out after administration. The untreated eyes served in each case as a control.

Slight redness, moderate chemosis and severely increased discharge along with the diffuse corneal opacity were observed in the eyes of the treated animals at 1h after application. Further, slight redness and diffuse areas of opacity persisted until 72 h post-application. However, these irritation response were completely reversible at one week after the treatment. Furthermore, no effects were observed in the iris during the study period.

Under the test conditions, the test substance was considered to be slightly irritating to eyes.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

A study was performed to assess the skin irritation potential of the test substance in male New Zealand White rabbits according to OECD Guideline 404, EPA OPPTS 870.2500 and EU Method B.4, in compliance with GLP. 500 mg of test substance moistened with water was applied to an approximately 6 cm² area of intact skin. Sterile gauze pads (5x5 cm) were placed on the skin of rabbits surrounded by an adhesive hypoallergenic plaster. The entire trunks of the animals were wrapped with plastic wrap for 4 h. After the exposure period, all remnants of test substance were carefully removed from the skin with body temperature water. Animals were scored for irritation reactions after 60 minutes, and at 24, 48 and 72 h. Except for very slight erythema at 24 h and 48 h, no other signs of irritation were observed in treated animals during the observation period. Also, the observed erythema was reversible with 72 h. Further, mean scores for dermal irritation for each animal were calculated to be 0.66 for erythema and 0.00 for edema formation. In addition, no clinical signs of systemic toxicity were observed. Under the test conditions, the test substance was considered to be slightly irritating to skin (Stahl, 2006a).

Eye irritation:

A single dose of 100 mg test substance was applied to the conjunctival sac of the left eye of each animal. The treated eyes were not washed out after administration. The untreated eyes served in each case as a control. Slight redness, moderate chemosis and severely increased discharge along with the diffuse corneal opacity were observed in the eyes of the treated animals at 1h after application. Further, slight redness and diffuse areas of opacity persisted until 72 h post-application. However, these irritation response were completely reversible at one week after the treatment. Furthermore, no effects were observed in the iris during the study period. Under the test conditions, the test substance was considered to be slightly irritating to eyes (Stahl, 2006b).


Justification for selection of skin irritation / corrosion endpoint:
Guideline-compliant study conducted according to GLP.

Justification for selection of eye irritation endpoint:
Guideline-compliant study conducted according to GLP.

Effects on eye irritation: slightly irritating

Justification for classification or non-classification

Skin irritation

Based on the results of a skin irritation study, the test substance does not need to be classified for skin irritation potential according to the EU CLP criteria (EC 1272/2008).

Eye irritation

Based on the results of an eye irritation study, the test substance is classified for eye irritation potential according to the EU CLP criteria (EC 1272/2008) as H319.