Registration Dossier

Evaluation of all animals (mean score)

Evaluation of each animal

Reference

Reference

According to results of an OECD TG 404 compliant study in the rabbit the test item is not irritating to the skin (reference 7.3.1 -1).

According to results of an OECD TG 405 compliant study in the rabbit the test item is not irritating to eyes (reference 7.3.2 -1).

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2004-10-29 to 2005-02-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Remarks:

rbt, Crl:KBL(NZW)BR

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 24 weeks
- Weight at study initiation: 4.96 kg
- 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
- Water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20°C
- Humidity: 39 - 62 %
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Remarks:

some drops of Aqua pro injectione to ensure good contact with the skin

Controls:

not required

Amount / concentration applied:

The test material was administered undiluted with 0.5 g onto 6 cm² patches.

Duration of treatment / exposure:

4 h

Observation period:

up to day 8

Number of animals:

3 females

Details on study design:

SCORING SYSTEM: according to Draize

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

(mean score 1)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

(mean score 2)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

(mean score 3)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

(mean score 1)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

(mean score 2)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

(mean score 3)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

After a single application to the intact dorsal skin of rabbits, for 4 hours under semiocclusive conditions, no signs of irritation were observed at the treated areas.

Other effects:

- Other adverse local effects: none
- Other adverse systemic effects: No signs of clinical toxicity were detected. All animals survived the observation period.

Table 1 individual local findings after 4 h exposure under semiocclusive conditions

Animal No.		Local findings on day
Animal No.		1	2	3	4	5	6	7	8
		(1 h)	(24 h)	(48 h)	(72 h)
15	Erythema	0	0	0	0	0	0	0	0
15	Edema	0	0	0	0	0	0	0	0
19	Erythema	0	0	0	0	0	0	0	0
19	Edema	0	0	0	0	0	0	0	0
21	Erythema	0	0	0	0	0	0	0	0
21	Edema	0	0	0	0	0	0	0	0

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study no skin irritation potential could be detected with the test item.

Executive summary:

The study was performed according to the OECD Guideline for Testing of Chemicals, No. 404, the annex to Directive 92/69 EEC, and the recommendations of DRAIZE (1959). To test for primary skin irritation, 0.5 g of the test material was mixed with some drops of Aqua pro injectione to ensure good contact with the skin. Afterwards the test material was spread onto 6 cm²patches and applied to the intact skin of previously shaven rabbits for a 4 hours period under semiocclusive conditions. The study was started as an initial test with one animal and followed by the confirmatory test with two further animals. The first examination of the treated skin sites followed 1 hour after removal of the patches. Thereafter, examinations were performed daily for further 7 days.
Under the conditions of the present study no signs of irritation were seen.

Evaluation of all animals (mean score)

Day (after treatment)	1 (1 hour)	2 (24 hours)	3 (48 hours)	4 (72 hours)
Erythema	0	0	0	0
Edema	0	0	0	0

Evaluation of each animal

Based on the results of this study, the test item is not considered to be irritating to skin.

Endpoint conclusion:

no adverse effect observed (not irritating)

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2004-11-18 to 2005-02-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Remarks:

rbt, Crl:KBL(NZW)

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age at study initiation: 28- 29 weeks
- Weight at study initiation: 4.78 kg
- 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
- Water:ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 17-18 °C
- Humidity: 46 - 60 %
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 / 12

Vehicle:

unchanged (no vehicle)

Remarks:

Before application, the test material was ground in a mortar using a pestle.

Controls:

not required

Amount / concentration applied:

100 mg

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3 females

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: fluorescein

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #1

Remarks:

(mean score 1)

Time point:

24/48/72 h

Score:

1.67

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #2

Remarks:

(mean score 2)

Time point:

24/48/72 h

Score:

1.67

Max. score:

3

Reversibility:

fully reversible within: 6 days

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #3

Remarks:

(mean score 3)

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 5 days

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

(mean score 1)

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

(mean score 2)

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

(mean score 3)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

(mean score 1)

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

(mean score 2)

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

(mean score 3)

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 5 days

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

(mean score 1)

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

(mean score 2)

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

(mean score 3)

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritant / corrosive response data:

Reversibility of any observed effect: Changes fully reversible within 7 days

Other effects:

No signs of irritation could be observed at the iris. The cornea showed scattered or diffuse areas of opacity (score 1) at the first reading up to day 4 of the experimental part. The conjunctivae showed redness (scores 1 and 2) and chemosis (scores 1 to 4) from the first reading up to experimental day 6, maximally. Furthermore discharge (scores 2 and 3) was seen from the first reading up to day 3.
Thereafter no signs of irritation were observed. The untreated eyes were unchanged.

Table 1 Individual findings

		Mean score (24, 48, and 72 hours)			Maximum value (24, 48, or 72 hours)
Animal No		17	18	20	17	18	20
Cornea	A	0.33	0.67	1.00	1	1	1
Iris	A	0.00	0.00	0.00	0	0	0
Conjunctivae	A	1.67	1.67	1.00	2	2	1
Conjunctivae	B	1.67	0.33	0.00	4	1	0

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of this study the test material is not regarded as irritant to the eyes.

Executive summary:

To test for primary eye irritation, 0.1 g of the solid test material was applied into the conjunctival sac of rabbits according to OECD TG 405. The first examination of the eyes followed 1 hour after instillation. The examinations were performed daily for further 7 days. No signs of irritation could be observed at the iris. The cornea showed scattered or diffuse areas of opacity (score 1) at the first reading up to day 4 of the experimental part. The conjunctivae showed redness (scores 1 and 2) and chemosis (scores 1 to 4) from the first reading up to experimental day 6, maximally. Furthermore discharge (scores 2 and 3) was seen from the first reading up to day 3. Thereafter no signs of irritation were observed. The untreated eyes were unchanged.

Day	1	2	3	4
(after treatment)	(1 hour)	(24 hours)	(48 hours)	(72 hours)
Cornea	1.00	1.00	0.67	0.33
Iris	0.00	0.00	0.00	0.00
Redness	1.33	1.67	0.33	0.00
Chemosis	3.00	1.67	0.33	0.00

Mean Score
(24, 48, 72 hours)

Max Score
(24, 48, 72 hours)

Animal No.

	Mean score			Max Score
	(1, 24, 48, 72 hours)			(1, 24, 48, 72 hours)
Animal No.	1	2	3	1	2	3
Cornea	0.33	0.67	1.00	1	1	1
Iris	0.00	0.00	0.00	0	0	0
Redness	1.67	1.67	1.00	2	2	1
Chemosis	1.67	0.33	0.00	4	1	0

Based on the results of this study the test material is not regarded as irritant to the eyes.

Endpoint conclusion:

no adverse effect observed (not irritating)

Endpoint conclusion:

no study available

Skin irritation, in vivo

The study was performed according to the OECD Guideline for Testing of Chemicals, No. 404, the annex to Directive 92/69 EEC, and the recommendations of DRAIZE (1959). To test for primary skin irritation, 0.5 g of the test material was mixed with some drops of Aqua pro injectione to ensure good contact with the skin. Afterwards the test material was spread onto 6 cm²patches and applied to the intact skin of previously shaven rabbits for a 4 hours period under semiocclusive conditions. The study was started as an initial test with one animal and followed by the confirmatory test with two further animals. The first examination of the treated skin sites followed 1 hour after removal of the patches. Thereafter, examinations were performed daily for further 7 days.
Under the conditions of the present study no signs of irritation were seen. Based on the results of this study, the test item is not considered to be irritating to skin.

Eye irritation, in vivo

To test for primary eye irritation, 0.1 g of the solid test material was applied into the conjunctival sac of rabbits according to OECD TG 405. The first examination of the eyes followed 1 hour after instillation. The examinations were performed daily for further 7 days. No signs of irritation could be observed at the iris. The cornea showed scattered or diffuse areas of opacity (score 1) at the first reading up to day 4 of the experimental part. The conjunctivae showed redness (scores 1 and 2) and chemosis (scores 1 to 4) from the first reading up to experimental day 6, maximally. Furthermore discharge (scores 2 and 3) was seen from the first reading up to day 3. Thereafter no signs of irritation were observed. The untreated eyes were unchanged. Based on the results of this study the test material is not regarded as irritant to the eyes.

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available in vivo data on skin and eye irritation, the test item is does not require classification as being irritant according to Regulation (EC) No 1272/2008 (CLP), as amended for the twelfth time in Regulation (EU) 2019/521.