Registration Dossier

Reference

Reference

Skin irritation in vivo (OECD TG 404): not irritating (reference 7.3.1-1).

Eye irritation in vivo (OECD TG 405): not irritating (reference 7.3.2-1).

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000-01-26 to 2000-02-02

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

July 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

29 Decembre 1992

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Kißlegg, Germany
- Age at study initiation: about 16 weeks
- Weight at study initiation: 3.27 kg (range from 3.20 to 3.36)
- Housing: separately in special rabbit cages (manufacturer: Becker; type K99/30 KU, floor area about 5400 cm², a shelter with an integrated sitting board of about 1820 cm²; overall height: 60 cm;)
- Diet: ad libitum, pellets (Altromin Standard Diet TPF(R) 2113)
- Water: ad libitum, tap water
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 22
- Humidity (%): 37 - 64
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL

Duration of treatment / exposure:

4 h

Observation period:

8 days

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: left side of the animals backs (3 patches of about 6 cm²
- Type of wrap: self-adhesive fabric (Fixomull® stretch, Beiersdorf)

REMOVAL OF TEST SUBSTANCE
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
1, 24, 48, 72 hours and then daily upt o study day 8

SCORING SYSTEM:
- Method of calculation: according to the DRAIZE-, OECD- and EEC recommendations

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No signs of irritation were observed in any of the 3 animals.

Interpretation of results:

GHS criteria not met

Conclusions:

According to the conducted study, the test substance is not regarded as irritant to the skin.

Executive summary:

The test on skin irritation was performed in accordance with OECD TG 404. The test substance (0.5 mL) was spread onto 6 cm² patches (3) and applied to the intact skin of 3 previously shaven New Zealand White rabbits (females) for a 4 hours period under semiocclusive conditions. The first examination of the treated skin sites followed 1 hour after removal of the patches. Thereafter, examinations were performed daily for further 8 days.

Under the conditions of the present study, no signs of toxicity were seen.

Evaluation of each animal:

No signs of irritation were observed in any of the 3 animals. Thus, the test material was not considered as a irritant to the skin.

Endpoint conclusion:

no adverse effect observed (not irritating)

Mean score (24h, 48h, 72h)

Maximum value (24h, 48h, or, 72h)

Animal No.

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000-01-26 to 2000-02-02

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

February 24, 1987

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

29 December 1992

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Kißlegg, Germany
- Age at study initiation: about 16 weeks
- Weight at study initiation: 3.41 kg (range from 3.34 to 3.52)
- Housing: separately in special rabbit cages (manufacturer: Becker; type K99/30 KU, floor area about 5400 cm², a shelter with an integrated sitting board of about 1820 cm²; overall height: 60 cm)
- Diet: ad libitum, pellets (Altromin Standard Diet TPFW 2113)
- Water: ad libitum, tap water
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 22
- Humidity (%): 37 - 64
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

other: adjacent eye served as control

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 mL

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

3

Details on study design:

SCORING SYSTEM:
CORNEA
Opacity: degree of density (area most dense taken for reading)
0 no ulceration or opacity
1 scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible
2 easily discernible translucent area, details of iris slightly obscured
3 nacreous areas, no details of iris visible, size of pupil barely discernible
4 Opaque cornea, iris not discernible through the opacity

IRIS
0 normal
1 markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive)
2 no reaction to light, haemorrhage, gross destruction (any or all of these)

CONJUNCTIVAE
Redness (refers to palpebral and bulbar conjunctivae, cornea and iris)
0 blood vessels normal
1 some blood vessels definitely hyperaemic (injected)
2 diffuse, crimson colour, individual vessels not easily discernible
3 diffuse beefy red

Chemosis: lids and/or nictitating membrane
0 no swelling
1 any swelling above normal (including 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

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No signs of irritation were observed in the cornea and iris. The conjunctivae showed redness (score 1) in two rabbits during hour one of the study. Furthermore no signs of irritation were observed. The untreated eyes were unchanged.

Interpretation of results:

GHS criteria not met

Conclusions:

According to the conducted study, the test substance is not regarded as irritant to the eyes.

Executive summary:

In a primary eye irritation study in accordance with OECD guideline 405 and EU method B.5, 0.1 mL of the undiluted test substance was instilled into the conjunctival sac of the right eye of three female New Zealand White rabbits. The eyes were not washed. Animals were then observed for up to 8 days. Irritation was scored by the method of Draize:

		Mean score (24, 48, and 72 hours)			Maximum value (24, 48, or 72 hours)
Animal No.		7	8	9	7	8	9
Cornea	A	0.00	0.00	0.00	0	0	0
Iris	A	0.00	0.00	0.00	0	0	0
Conjunctivae	A	0.00	0.00	0.00	0	0	0
	B	0.00	0.00	0.00	0	0	0

No signs of irritation were observed in the cornea and iris. The conjunctivae showed redness (score 1) in two rabbits during hour one of the study. Furthermore no signs of irritation were observed. The untreated eyes were unchanged. Based on the above results the test substance was non-irritating to eyes.

Endpoint conclusion:

no adverse effect observed (not irritating)

Endpoint conclusion:

no study available

Skin irritation

The test on skin irritation was performed in accordance with OECD TG 404. The test substance (0.5 mL) was spread onto 6 cm² patches (3) and applied to the intact skin of 3 previously shaven New Zealand White rabbits (females) for a 4 hours period under semiocclusive conditions. The first examination of the treated skin sites followed 1 hour after removal of the patches. Thereafter, examinations were performed daily for further 8 days.

Under the conditions of the present study, no signs of toxicity were seen.

Evaluation of each animal:

No signs of irritation were observed in any of the 3 animals. Thus, the test material was not considered as a irritant to the skin.

Eye irritation

In a primary eye irritation study in accordance with OECD guideline 405 and EU method B.5, 0.1 mL of the undiluted test substance was instilled into the conjunctival sac of the right eye of three female New Zealand White rabbits. The eyes were not washed. Animals were then observed for up to 8 days. Irritation was scored by the method of Draize:

		Mean score (24, 48, and 72 hours)			Maximum value (24, 48, or 72 hours)
Animal No.		7	8	9	7	8	9
Cornea	A	0	0	0	0	0	0
Iris	A	0	0	0	0	0	0
Conjunctivae	A	0	0	0	0	0	0
	B	0	0	0	0	0	0

No signs of irritation were observed in the cornea and iris. The conjunctivae showed redness (score 1) in two rabbits during hour one of the study. Furthermore no signs of irritation were observed. The untreated eyes were unchanged. Based on the above results the test substance was non-irritating to eyes.

The available experimental test data is reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result the substance is considered not to be classified for eye or skin irritation under Regulation (EC) No 1272/2008, as amended for the twelfth time in Regulation (EU) No 2019/521.