Registration Dossier

Administrative data

Description of key information

Not skin irritating
Not eye irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation / corrosion, other
Remarks:
An in vitro skin irritation study does not need to be conducted because adequate data from an in vivo study are available
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
January from 10th to 24th, 1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study conducted according to internationally accepted testing guidelines. Justification for Read Across is detailed in the endpoint summary.
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
Adopted 17.7.1992
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Interfauna U.K. Ltd., Wyton, Huntingdon, England. The health of the animals was routinely examined for the main specific Pathogens by the breeder.
- Age at study initiation: healthy adult albino rabbits.
- Weight at study initiation: 3.8 - 4.1 kg
- Housing: individually housed in stainless steel cages with flat rod bases or plastic cages with perforated bases, under standardized conventional conditions. Excrement trays beneath the cages contained low-dust (wood) bedding (type S 8/15, Ssniff Spazialdiaten GmbH, Soest). Bedding was regularly spot-checked for contaminants as required by the Department of Laboratory Animal Services, and changed at least twice weekly.
- Diet: standard diet "Ssniff K4" (ssniff Spezialdiaten GmbH, Soest), approx. 100 - 120 g per animal/day; once per day in the morning.
- Water: tap water (of drinking water quality), ad libitum.
- Acclimation period: prior to use the animals were acclimatized for approximately one week in the animal room.
- Health control: no vaccinations or treatment with antibiotics were performed prior to receipt of the animals, or during the acclimatization phase or study period. If females were used they were nulliparous and not pregnant. The animals were examined one day prior to the start of the
study to establish that they were biologically normal; only animals not exhibiting any alterations to skin or eyes were used.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 1.5 °C (higher when ambient temperature over 24 °C.
- Humidity: 50 - 70 %
- Air changes: 12 - 15 times per hour.
- Photoperiod: 12 hours light/dark cycle, artificial illumination from 6 a.m. to 6 p.m.; approx. 500 Lux.
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
water
Controls:
yes, concurrent vehicle
Amount / concentration applied:
TEST MATERIAL
500 ng of the pulverized test substance were moistened with deionised water (to ensure good contact with the skin) and subsequently applied to a hypoallergenic Hansamed - patch (Beiersdorf No. 2342 PV3).

CONTROL
A further patch was moistened with water. The patch prepared in this way was placed on the opposite dorso-lateral area of the trunk of each animal respect to the test item one.
Duration of treatment / exposure:
Four hours
Observation period:
14 days
Number of animals:
Three
Details on study design:
TEST SITE
- Area of exposure: approximately 24 hours before the test, fur was shorn from the dorso-lateral area of the trunk (6 x 6 cm) of each of the rabbits (equipment: electric hair clipper). Care was taken to avoid abrading the skin. Only animals with healthy and intact skin were used.

REMOVAL OF TEST SUBSTANCE
- Washing: the exposed skin areas were carefully washed with water without altering the existing response, or the integrity of the epideinis. The contralateral skin area not treated with test substance be served as control.
- Time after start of exposure: 4 hours

SCORING SYSTEM
For each animal the Draize scores recorded approx. 24, 48, and 72 hours after application were recorded.

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

Oedema Formation
No oedema: 0
Very slight oedema (barely perceptible): 1
Slight oedema (edges of area well defined by definite raising): 2
Moderate oedema (raised approximately 1 mm): 3
Severe oedema (raised more than 1 mm and extending beyond area of exposure): 4
Irritation parameter:
edema score
Basis:
animal: 3/3
Time point:
other: mean at 24, 48 and 72 hrs
Score:
< 2.3
Reversibility:
fully reversible
Irritant / corrosive response data:
Evaluation of skin erythema was not possible in all three animals due to the intense colouration by the test substance.
Nevertheless, no other inflammatory signs (eschar or oedema formation) became apparent within the observation period of 7 days.

Signs of toxicity were not observed. This evidence indicates no hazard potential to the skin and the test substance may therefore be regarded as ‘not irritating to the skin'.

Individual findings

Animals N. Body weight Draize grade after
1 h 24 h 48 h 72 h 7 d 14 d
E O E O E O E O E O E O
E2 4.1 kg a 0 a 0 a 0 a 0 a 0 - -
F16 3.8 kg a 0 a 0 a 0 a 0 a 0 - -
G9 3.8 kg a 0 a 0 a 0 a 0 a 0 - -

E: Erythema

O: Oedema

a: exposed skin areas stained in colour of the test substance, evaluation of erythema not possible

Interpretation of results:
not irritating
Remarks:
Migrated information
Conclusions:
Not irritating
Executive summary:

Tests for the determination of irritant/corrosive effects of the substance on skin of albino rabbits were conducted in accordance with the OECD Guidelines Nos. 404. The albino rabbit is recommended as the preferred species in the guideline.

Evaluation of skin erythema was not possible in all three animals due to the intense colouration by the test substance. Nevertheless, no other inflammatory signs (eschar or oedema formation) became apparent within the observation period of 7 days.

Signs of toxicity were not observed. This evidence indicates no hazard potential to the skin and the test substance may therefore be regarded as ‘not irritating to the skin'.

Conclusion

Not irritating

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation, other
Remarks:
An in vitro eye irritation study does not need to be conducted because adequate data from an in vivo study are available
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
January from 10th to 24th, 1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study conducted according to internationally accepted testing guidelines. Justification for Read Across is detailed in the endpoint summary.
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
Adopted 24.2.1987
Deviations:
yes
Remarks:
more accurate description and scoring of corneal defects, additional examination of aqueous humour
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Interfauna U.K. Ltd., Wyton, Huntingdon, England. The health of the animals was routinely examined for the main specific Pathogens by the breeder.
- Age at study initiation: healthy adult albino rabbits.
- Weight at study initiation: 3.5 - 4.0 kg
- Housing: individually housed in stainless steel cages with flat rod bases or plastic cages with perforated bases, under standardized conventional conditions. Excrement trays beneath the cages contained low-dust (wood) bedding (type S 8/15, Ssniff Spazialdiaten GmbH, Soest). Bedding was regularly spot-checked for contaminants as required by the Department of Laboratory Animal Services, and changed at least twice weekly.
- Diet: standard diet "Ssniff K4" (ssniff Spezialdiaten GmbH, Soest), approx. 100 - 120 g per animal/day; once per day in the morning.
- Water: tap water (of drinking water quality), ad libitum.
- Acclimation period: prior to use the animals were acclimatized for approximately one week in the animal room.
- Health control: no vaccinations or treatment with antibiotics were performed prior to receipt of the animals, or during the acclimatization phase or study period. If females were used they were nulliparous and not pregnant. The animals were examined one day prior to the start of the
study to establish that they were biologically normal; only animals not exhibiting any alterations to skin or eyes were used.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 1.5 °C (higher when ambient temperature over 24 °C.
- Humidity: 50 - 70 %
- Air changes: 12 - 15 times per hour.
- Photoperiod: 12 hours light/dark cycle, artificial illumination from 6 a.m. to 6 p.m.; approx. 500 Lux.
Vehicle:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount applied: after gently pulling the lower lid away from the eyeball a volume of 100 µl of the pulverized test substance - equivalent
to approx. 67 mg — was placed int the conjunctival sac of one eye of each of three rabbits. The lids were then gently held together for about one second in order to limit loss of the material.
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
Three rabbits
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: the treated eye was rinsed with normal saline.
- Time after start of exposure: 24 hours after instillation.

TOOL USED TO ASSESS SCORE: hand-slit lamp. To define epithelial damage, one drop of a 1 % fluorescein solution was applied to the corneal surface 24 hours after administration of the test substance. The eye was then rinsed with normal saline to remove excess and nonabsorbed fluroscein. Evaluation was performed by means of ultraviolet illumination (area), in darkened room and diffuse white illumination (intensity), according to McDonald and Shadduck.

SCORING SYSTEM
The signs of cornea (opacity and area affected), iris (hyperaemia, reaction to light), conjunctivae, i.e. conjunctiva of bulbus, lids, and nictitating membrane (erythema, chemosis), and discharge were recorded as described by Draize and the aqueous humor (opacity) as described by McDonald and Shadduck.

A) Cornea
Opacity and degree of density (most dense area scored) .
No opacity: 0
Scattered or diffuse area, details of iris clearly visible: 1
Easily discernible translucent areas, details of iris slightly obscured: 2
Opalescent areas, no details of iris visible, size of pupil barely discernible: 3
Opaque, iris invisible: 4

Area of cornea involved
One quarter (or less) but not zero: 1
Greater than one quarter, but less than half: 2
Greater than half, but less than three quarters: 3
Greater than three quarters, up to whole area: 4

B) Fluorescein Staininq (Epithelial Defects)
Intensity
- absence of staining 0
- slight staining, underlying structures easily visible 1
- moderate staining, underlying structures easily visible, although there is some loss of detail 2
- marked staining, underlying structures barely visible but not completely obliterated 3
- extreme staining, underlying structures cannot he observed 4

To assess the area the criteria of A) were used. In addition, differentiation was made between scattered punctate and confluent diffuse areas.

C) Iritis
A Values
Normal: 0
Folds above normal, congestion, swelling, circumcornea injection (any or all of these or combination of any thereof) iris still reacting to light (sluggish reaction is positive: 1
No reaction to light, hemorrhage, gross destruction (any or all of these): 2

D) Aqueous Flare
- light beam in the anterior chamber not visible (no Tyndall effect) 1
- Tyndall effect barely discernible, intensity of light beam in the anterior chamber less than intensity of light passing through lens 2
- Tyndall effect in the anterior chamber easily discernible, intensity of light beam equal to slit beam passing through lens 2
- Tyndall effect easily discernible, intensity of light beam greater than slit beam passing through lens 3
Tyndall phenomenon: Scattefing illumination arising when a collected light beam passes through a colloidal solution.

E) Conjunctivae
Erythema (redness)
Blood vessels normal: 0
Some blood vessels definitely injected above normal: 1
More diffuse, deeper crimson red, individual vessels not easily discernible: 2
Diffuse beefy red: 3

Chemosis
No swelling: 0
Any swelling above normal (includes nictitating membrane): 1
Obvious swelling with partial eversion of lids: 2
Swelling with lids about half closed: 3
Swelling with lids about half closed to completely closed: 4

F) Discharge
No discharge: 0
Any amount different from normal (does not include small amounts observed in inner cantus of normal animals): 1
Discharge with moistening of the lids and hairs just adjacent to lids: 2
Discharge with moistening of the lids and hairs, and considerable area around the eye: 3
Irritation parameter:
cornea opacity score
Basis:
animal: 3/3
Time point:
other: mean at 24, 48 and 72 hrs
Score:
< 1
Irritation parameter:
iris score
Basis:
animal: 3/3
Time point:
other: mean at 24, 48 and 72 hrs
Score:
< 1
Irritation parameter:
conjunctivae score
Basis:
animal: 3/3
Time point:
other: mean at 24, 48 and 72 hrs
Score:
< 2
Reversibility:
fully reversible
Irritation parameter:
chemosis score
Basis:
animal: 3/3
Time point:
other: mean at 24, 48 and 72 hrs
Score:
< 2

Individual findings at the various observation time

Animals N. Body weight Tissue Signs Draize grade
1h 24 h 48 h 72 h 7 d 14 d 21 d
F12 3.9 kg cornea o 0 0 0 0 0 - -
a 0 0 0 0 0 - -
fluorescein i - 0 - - - - -
a - 0 - - - - -
iris 0 0 0 0 0 - -
conjunctivae r e e 1f 0f 0f - -
c 1 0 0 0 0 - -
aqueous humor 0 0 0 0 0 - -
discharge 2 0 0 0 0 - -
G5 3.5 kg cornea o 0 0 0 0 0 - -
a 0 0 0 0 0 - -
fluorescein i - 0 - - - - -
a - 0 - - - - -
iris 0 0 0 0 0 - -
conjunctivae r e 1f 1f 0f 0f - -
c 0 0 0 0 0 - -
aqueous humor 0 0 0 0 0 - -
discharge 1 0 0 0 0 - -
E4 4.0 kg cornea o 0 0 0 0 0 - -
a 0 0 0 0 0 - -
fluorescein i - 0 - - - - -
a - 0 - - - - -
iris 0 0 0 0 0 - -
conjunctivae r e 1f 1f 0f 0f - -
c 0 0 0 0 0 - -
aqueous humor 0 0 0 0 0 - -
discharge 1 0 0 0 0 - -

-: not examined; o: opacity; a: area; e: evaluation not possible due to the colouration by the test substance; f: colouration by the test substance, evaluation possible; i: intensity; r: redness; c: chemosis

Interpretation of results:
not classified
Remarks:
Migrated information according to the CLP Regulation (EC 1272/2008) Criteria used for interpretation of results: EU
Conclusions:
The substance does not meet the criteria to be classified as eye irritating, according to the CLP Regulation (EC 1272/2008).
Executive summary:

Tests for the determination of irritant/corrosive effects of the substance on eyes of albino rabbits were conducted in accordance with the OECD Guidelines Nos. 405. The albino rabbit is recommended as the preferred species in these guidelines. Slight deviations from the guideline (e.g. more accurate description and scoring of corneal defects, additional examination of aqueous humour) do not affect the validity of this study.

A single application of 100 µl (67 mg) of the test material was administered to eye of three rabbits. 24 hours after instillation the treated eye was rinsed with normal saline. Ocular reactions were recorde 1, 24, 48 and 72 hours after administration.

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for iritis, lower than 2 for both conjunctival redness and oedema, in all animals.

Conclusion

The substance does not meet the criteria to be classified as eye irritating, according to the CLP Regulation (EC 1272/2008).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

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), 3.2 Skin corrosion/irritation section, skin Irritation means the production of reversible damage to the skin following the application of a test substance for up to 4 hours.

In the study performed on the structural analogous, the mean values from gradings at 24, 48 and 72 hours after patch removal were lower than 2.3 in at least 2 of 3 animals for oedema reactions. Evaluation of skin erythema was not possible in all three animals due to the intense colouration by the test substance.

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.

In the study performed on the structural analogous, the mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for iritis, lower than 2 for both conjunctival redness and oedema, in all tested animals.

In conclusion, the substance under registration is expected to not meet the criteria to be classified for the eye/skin irritation, according to the CLP Regulation (EC 1272/2008).