Registration Dossier

Administrative data

Description of key information

Not skin irritant

Irreversible effects on the eye

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Justification for type of information:
Details in section 13
Reason / purpose:
read-across source
Reason / purpose:
read-across: supporting information
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
1 males and 2 females
- Body weight at start of acclimatization: 2.9 kg (male) and 2.6-2.8 kg (females)
- Body weight at start of treatment: 3.2 kg (male) and 2.8-3.1 kg (females)
- Age at treatment: 15 weeks
- Acclimatization: 4 days under test conditions after health examination
- Accomodation: individually in stainless steel cages with an automatic cleaning system, equipped with feed hoppers, drinking water bowls and wood for gnawing- Diet: pelleted standard Kliba 341 rabbit maintenance diet (batch 94/96) ad libitum from Kliba Mühlen AG, CH-4303 Kaiseraugst
- Water: community tap water from Itingen, ad libitum
- Standard laboratory conditions: Air-conditioned with 10-15 air changes per hour and continuously monitored environment with target ranges for temperature of 20 ± 3 °C and for relative humidity between 40-70 % (values above 70 % during cleaning process possible), 12 hours artificial fluorescent light/12 hours dark, music during the light period.
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
water
Remarks:
bi-distilled
Amount / concentration applied:
0.5 g of test substance
Duration of treatment / exposure:
4 hours of application
Observation period:
72 hours
Number of animals:
3 (1 male and 2 females)
Details on study design:
TREATMENT
Approximately 24 hours before treatment, the dorsal fur was clipped with an electric clipper, exposing an area of approximately 100 square centimeters (10 cm x 10 cm). The skin of the animals was examined once, 1 day prior to test substance administration.Only those animals with no signs of skin injury or irritation were used in the test.On the day of treatment, 0.5 g of the test article was applied to approx. 6 cm2 of the intact skin of the clipped area. It was covered with a 2.5 cm x 2.5 cm patch of surgical gauze. The gauze was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with an elastic bandage. The test substance was applied in such a position as to preclude oral ingestion after removal of the dressing.The duration of treatment was 4 hours. Then the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site so that any reactions (erythema) were clearly visible at that time.

SCORING SYSTEM

Erythema and Eschar Formation
No erythema:0
Very slight erythema (barely perceptible): 1
Well defined erythema: 2
Moderate to severe erythema: 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth): 4

Edema Formation
No edema: 0
Very slight edema (barely perceptible): 1
Slight edema (edges of area well defined by definite raising): 2
Moderate edema (raised approximately 1 mm): 3
Severe edema (raised more than 1 mm and extending beyond area of exposure): 4
Irritation parameter:
erythema score
Basis:
animal: all animals (3/3)
Time point:
72 h
Score:
0
Remarks on result:
not determinable
Remarks:
after 24 and 48 h the evaluation of some parameters was precluded due to staining.
Irritation parameter:
erythema score
Basis:
mean
Time point:
24/48/72 h
Remarks on result:
not determinable
Irritation parameter:
edema score
Basis:
animal: all animals (3/3)
Time point:
24/48/72 h
Score:
0
Irritant / corrosive response data:
The substance showed a primary irritation score of 0 when applied to healthy intact rabbit skin.
Local signs (mean values from 24 to 72 hours) consisted of grade 0 erythema and grade 0 edema.
Blue or dark blue staining by the test article of the treated skin was observed.
No irreversible alterations of the treated skin were observed nor were corrosive effects evident on the skin.
Other effects:
No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred.The body weight gain of all rabbits was within the normal range of variability.
Interpretation of results:
other: not classified according to the CLP Regulation
Conclusions:
The test substance was found to be non irritant when applied to intact rabbit skin.
Executive summary:

The skin irritation test was performed to rabbit skin according to the OECD guideline 404.

The test substance was found to be non irritant when applied to intact rabbit skin: local signs (mean values from 24 to 72 hours) consisted of grade 0 erythema and grade 0 edema. The test article caused blue or pale blue staining of the treated skin, therefore the erythema reaction could not be evaluated after 24 and 48 hours, nevertheless the possible score are not considered to be significant . No corrosive effects were noted on the treated skin of any animal at any measuring interval.

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:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Justification for type of information:
Details in section 13
Reason / purpose:
read-across source
Reason / purpose:
read-across: supporting information
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
1 males and 2 females
- Body weight at start of acclimatization: 2.7 kg (male) and 2.8-3.0 kg (females)
- Body weight at start of treatment: 2.9 kg (male) and 2.9-3.0 kg (females)
- Age at treatment: 15 weeks
- Acclimatization: 4 days under test conditions after health examination
- Accomodation: individually in stainless steel cages with an automatic cleaning system, equipped with feed hoppers, drinking water bowls and wood for gnawing
- Diet: pelleted standard Kliba 341 rabbit maintenance diet (batch 94/96) ad libitum from Kliba Mühlen AG, CH-4303 Kaiseraugst
- Water: community tap water from Itingen, ad libitum
- Standard laboratory conditions: Air-conditioned with 10-15 air changes per hour and continuously monitored environment with target ranges for temperature of 20 ± 3 °C and for relative humidity between 40-70 % (values above 70 % during cleaning process possible), 12 hours artificial fluorescent light/12 hours dark, music during the light period.
Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
0.1 g of test substance (undiluted)
Duration of treatment / exposure:
single apllication to the conjunctival sac of the left eyelid
Observation period (in vivo):
21 days
Number of animals or in vitro replicates:
1 male and 2 females
Details on study design:
TREATMENT:
The eyes of the animals were examined for intactness once, one day prior to test substance administration.Only those animals with no signs of ocular injury or irritation were used on the test.At treatment, 0.1 g of the test article (undiluted) was placed in the conjunctival sac of the left eye of each animal 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 right eye remained untreated and served as the reference control.

SCORING SYSTEM:
CORNEAL IRRITATION
Opacity: degree of density (densest area used for assessment)
No ulceration or opacity........................................................................................................................................... 0
Scattered or diffuse areas of opacity (other than slight dulling of normal luster), details of iris clearly visible ......1
Easily discernible translucent area, details of iris slightly obscured ........................................................................2
Nacrous area, no details of iris visible, size of pupil barely discernible ..................................................................3
Opaque cornea, iris not discernible through the opacity .........................................................................................4

IRIDIC IRRITATION
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperemia, or injection, any of these or combination thereof, iris still reacting to light (sluggish reaction is positive) .................................................................................................................................................................................1
No reaction to light, hemorrhage, gross destruction (any or al 1 of these)............................................................ 2

CONJUNCTIVAL IRRITATION
Redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris).
Blood vessels normal.............................................................................................................................................. 0
Some blood vessels definitely hyperemic (injected)............................................................................................... 1
Diffuse, crimson color, individual vessels not easily discernible............................................................................ 2
Diffuse beefy red ....................................................................................................................................................3

Chemosis:
lids and/or nictitating membranes No swelling ..........................................................................................................0
Any swelling above normal (includes nictitating membranes)...................................................................................1
Obvious swelling with partial eversion of lids ..........................................................................................................2
Swel 1 i ng wi th 1 i ds about hal f cl osed ...............................................................................................................3
Swelling with lids more than half closed ...................................................................................................................4

The maximum scores are:
Cornea : 4 Conjunctival Redness : 3 Iris : 2 Conjunctival Chemosis : 4
Maximum attainable score : 13

When present, corrosion of and/or staining of sclera and cornea by the test
article were recorded and reported

READINGS: 1h, 24h, 48h, 72h, 8d, 15d and 21d after application
Irritation parameter:
cornea opacity score
Basis:
animal: all animals (24, 25 and 26)
Time point:
24/48/72 h
Score:
0
Reversibility:
other: no ocular effects observed
Irritation parameter:
iris score
Basis:
animal: all animals (24, 25 and 26)
Time point:
24/48/72 h
Score:
0
Reversibility:
other: no ocular effects observed
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal: 24
Time point:
24/48/72 h
Score:
ca. 2
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal: 25
Time point:
24/48/72 h
Score:
ca. 1.67
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal: 26
Time point:
24/48/72 h
Score:
ca. 2
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
chemosis score
Basis:
animal: 24
Time point:
24/48/72 h
Score:
ca. 2.3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal: 25
Time point:
24/48/72 h
Score:
ca. 1.33
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal: 26
Time point:
24/48/72 h
Score:
ca. 2
Reversibility:
not fully reversible within: 21 days
Irritant / corrosive response data:
Reddening and swelling of the conjunctivae was evident in all animals, as was watery discharge. Hyperemia of the scleral blood vessels was noted in two animals. Reddening and swelling of the conjunctivae persisted in one animal until the end of observation, whereas reddening of the conjunctivae was noted ina second animal. Blue or light blue staining of the treated eyes by the test article was observed.No corrosion was observed at any of the measuring intervals.
Other effects:
The body weight of the animals was within the normal range of variability.
Interpretation of results:
other: Irreversible effects on the eye (Category 1) according to the CLP Regulation
Conclusions:
The test substance produced irreversible effects on the eye.
Executive summary:

The eye irritation test was performed to rabbit eye according to the OECD Guyideline 405.

The substance was found to cause irritation: in particular, the mean values at 24, 48 and 72 hours for not rinsed eyes were 0 for corneal opacity and iris in all animals, equal or higher than 2 for both conjunctival redness and chemosis, in two of three animals tested. In addition, the irritation symptoms were not fully reversible after 21 days of observation. Blue or light blue staining of the treated eyes by the test article was observed.

Therefore, the test substance can be classified in Category 1 due to irreversible effects on the eye, according to the CLP Regulation (EC n. 1272/2008).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

According to the CLP Regulation (EC 1272/2008) substances that have the potential to induce reversible damage to the skin following the application of a test substance for up to 4 hours are classified in Category 2 (irritating to the skin) if:

- Mean value of ≥ 2.3 ≤ 4.0 for erythema/eschar or for oedema in at least 2 of 3 tested animals from gradings at 24, 48 and 72 hours after patch removal or, if reactions are delayed, from grades on 3 consecutive days after the onset of skin reactions; or

- Inflammation that persists to the end of the observation period normally 14 days in at least 2 animals, particularly taking into account alopecia (limited area), hyperkeratosis, hyperplasia, and scaling; or

- In some cases where there is pronounced variability of response among animals, with very definite positive effects related to chemical exposure in a single animal but less than the criteria above.

In the skin irritation test of the similar substance 01, the mean values at 24, 48 and 72 hours after patch removal were lower than 2.3 (explicitly, the mean values were zero) in all animals for both erythema/eschar and oedema reactions (in both intact and abraded skin). In the skin irritation test of the substance, the mean values at 24, 48 and 72 hours after patch removal were lower than 2.3 (explicitly, the mean values were zero) in all animals for edema reactions (in both intact and abraded skin).

According to the CLP Regulation (EC 1272/2008) substances that have the potential to induce reversible eye irritation are classified in Category 2 (irritating to eyes) if, when applied to the eye of an animal, a substance produces at least in 2 of 3 tested animals, a positive response of:

- corneal opacity ≥ 1 and/or

- iritis ≥ 1, and/or

- conjunctival redness ≥ 2 and/or

- conjunctival oedema (chemosis) ≥ 2,

calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material, and which fully reverses within an observation period of 21 days.

The results show that the substance is not classified for skin irritation.

The substance was found to cause irritation in not rinsed eyes after 21 day of observation, especially in terms of conjunctival redness and chemosis.

Therefore, the test substance can be classified in Category 1 due to irreversible effects on the eye, according to the CLP Regulation (EC n. 1272/2008).