Registration Dossier

SKIN IRRITATION
Irritating, New Zealand White, United States EPA, Paragraph 163.81-5 “Primary dermal irritation study”, Federal Register, Vol. 43 No. 163, August 22, 1978, Seifert 1982a
EYE IRRITATION
Irritating, New Zealand White, United States EPA, Paragraph 163.81-4 “Primary eye irritation study”, Federal Register, Vol. 43 No. 163, August 22, 1978, Seifert 1982b

Reference

Endpoint:

skin irritation / corrosion, other

Remarks:

According to OECD 404

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 October 1982 to 25 October 1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP in accordance with generally accepted scientific principles, possibly with incomplete reporting or methodological deficiencies, which do not affect the quality of the relevant results.

Qualifier:

equivalent or similar to guideline

Guideline:

other: Procedure similar to the proposed guidelines of the United States EPA, Paragraph 163.81-5 “Primary dermal irritation study”, Federal Register, Vol. 43 No. 163, August 22, 1978

Deviations:

yes

Remarks:

: the procedure was based upon the guideline but was not in every part identical.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

: the animals were exposed to the test material for 24 hours in an occlusive fashion; there was no evaluation 60 minutes after patch removal and the observation period lasted only 7 days.

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2 - 3 kg
- Housing: Individually in metal cages.
- Diet (e.g. ad libitum): Rabbit food, ad libitum.
- Water (e.g. ad libitum): ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 55 ± 15 %
- Air changes (per hr): Approximately 15 per hour.
- Photoperiod (hrs dark / hrs light): 12 hours light per day.

IN-LIFE DATES: From: 18 October 1982 To: 25 October 1982

Type of coverage:

occlusive

Preparation of test site:

other: shaved and abraded

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

- Volume: 0.5 mL of the test material

Duration of treatment / exposure:

24 hours

Observation period:

7 days

Number of animals:

6; 3 male and 3 female

Details on study design:

SITE PREPARATION:
-24 hours before exposure, the flanks of the test animals were shaved using an electric clipper. Immediately prior to treatment, the skin on one side was slightly scarified by means of a "Schroepfschnaepper".

TEST SITE
- Patch Size: Gauze patches 2.5 x 2.5 cm soaked with the test material were applied to intact and abraded skin sites. The patches were covered with an impermeable material and were fastened to the body of the rabbit with adhesive tape.

SCORING SYSTEM: The skin reactions were assessed upon patch removal and during the subsequent 7 day observation period in accordance with the Draize scale (1977).

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Remarks:

of 6 animals

Time point:

other: mean of 24 and 72 hour evaluations

Score:

5.6

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Both shaved and abraded skin

Irritation parameter:

erythema score

Basis:

mean

Remarks:

of 6 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

3

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Shaved skin

Irritation parameter:

erythema score

Basis:

mean

Remarks:

of 6 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

2.9

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Abraded skin

Irritation parameter:

edema score

Basis:

mean

Remarks:

of 6 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

2.7

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Shaved skin

Irritation parameter:

edema score

Basis:

mean

Remarks:

of 6 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

2.6

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Abraded skin

Irritant / corrosive response data:

Necrosis and extended erythema was observed in all 6 animals.

Table 1 PII at 24 and 72 hours.

PII = total ÷ 4

Time After Exposure (hours)	Mean Erythema Scores		Mean Oedema Scores
Time After Exposure (hours)	Intact Skin	Abraded Skin	Intact Skin	Abraded Skin
24	3.0	3.0	2.8	2.7
72	3.0	2.8	2.7	2.5

Subtotal	6.0	5.8	5.5	5.2
Total	22.5
PII	5.6

Table 2 Individual Scores

Shaved

	Time After Exposure (hours)	Male			Female			Average
	Time After Exposure (hours)	1	2	3	1	2	3
Erythema	24 hours	3	3	3	3	3	3	3.0
	48 hours	3	3	3	3	3	3	3.0
	72 hours	3	3	3	3	3	3	3.0
	4 days	3	3	3	3	3	3	3.0
	7 days	3	2	3	2	2	3	2.5
Oedema	24 hours	3	3	3	2	3	3	2.8
	48 hours	3	2	2	2	3	3	2.5
	72 hours	4	2	2	2	3	3	2.7
	4 days	4	2	2	2	3	3	2.7
	7 days	4	2	2	2	2	2	2.3
		Abraded
Erythema	24 hours	3	3	3	3	3	3	3.0
	48 hours	2	3	3	3	3	3	2.8
	72 hours	2	3	3	3	3	3	2.8
	4 days	2	3	3	3	3	3	2.8
	7 days	2	2	3	2	2	2	2.2
Oedema	24 hours	2	3	3	2	3	3	2.7
	48 hours	2	2	3	2	3	3	2.5
	72 hours	2	2	4	2	3	2	2.5
	4 days	2	2	4	2	3	2	2.5
	7 days	2	2	3	1	2	2	2.0

Interpretation of results:

Category 2 (irritant)

Remarks:

Classified according to EU criteria.

Conclusions:

Under the conditions of the study, the test material was shown to cause marked irritation to the skin of rabbits and as such is classified as a category 2 skin irritant in accordance with EU criteria.

Executive summary:

The skin irritancy potential of the test material was assessed in a GLP compliant in vivo study that was based upon the procedure outlined in the guidelines of the United States EPA, Paragraph 163.81-5 “Primary dermal irritation study”, Federal Register, Vol. 43 No. 163, August 22, 1978.

Six New Zealand White rabbits were exposed to the test material in an occlusive fashion for 24 hours on both intact and abraded skin. The skin reactions were evaluated in accordance with the Draize scale at 24, 48 and 72 hours followed by observations after 4 and 7 days.

Under the conditions of this study, the test material produced marked irritation to the skin and as such requires classification as “Category 2, H315: Causes skin irritation” with signal word “Warning” in accordance with EU criteria.

Endpoint conclusion:

adverse effect observed (irritating)

Reference

Endpoint:

eye irritation, other

Remarks:

According to OECD 405

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 October 1982 to 25 October 1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP in accordance with generally accepted scientific principles, possibly with incomplete reporting or methodological deficiencies, which do not affect the quality of the relevant results.

Qualifier:

equivalent or similar to guideline

Guideline:

other: Procedure similar to the proposed guidelines of the United States EPA, Paragraph 163.81-4 “Primary eye irritation study”, Federal Register, Vol. 43 No. 163, August 22, 1978

Deviations:

yes

Remarks:

The procedure was based upon the guideline but was not in every part identical.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

The animals were not evaluated 60 minutes after exposure and the observation period lasted only 7 days.

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2 - 3 kg
- Housing: Individually in metal cages.
- Diet (e.g. ad libitum): Rabbit food, ad libitum.
- Water (e.g. ad libitum): ad libitum.
- Acclimation period: 4 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 55 ± 15 %
- Air changes (per hr): Approximately 15 per hour.
- Photoperiod (hrs dark / hrs light): 12 hours light per day.

IN-LIFE DATES: From: 18 October 1982 To: 25 October 1982

Vehicle:

unchanged (no vehicle)

Controls:

other: The right eye of each animal remained untreated and served as concurrent control.

Amount / concentration applied:

- Volume: 0.1 mL

Duration of treatment / exposure:

After insertion of the test material into the conjunctival sac, the eyelids were gently closed for 15 seconds.

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

9; 6 male and 3 female

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): in 3 of the 9 rabbits the eye was flushed with 10 mL of sterile physiological saline
- Time after start of exposure: approximately 30 seconds after treatment

SCORING SYSTEM:
- Eyes were scored according to the Draize scale (1977)

TOOL USED TO ASSESS SCORE: slit-lamp

Irritation parameter:

other: PII

Basis:

mean

Time point:

other: 7 days

Score:

29.6

Remarks on result:

other: Unrinsed eyes

Irritation parameter:

other: PII

Basis:

mean

Time point:

other: 7 days

Score:

22.1

Remarks on result:

other: Rinsed eyes

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 6 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

1

Max. score:

1

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Unrinsed eyes

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

1

Max. score:

1

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Rinsed eyes

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 6 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

0.5

Max. score:

1

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Unrinsed eyes

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

other: Rinsed eyes

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

of 6 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

2.9

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Unrinsed eyes

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

3

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Rinsed eyes

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 6 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

2.8

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Unrinsed eyes

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

2.7

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Rinsed eyes

Irritation parameter:

other: discharge

Basis:

mean

Remarks:

of 6 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

2.7

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Unrinsed eyes

Irritation parameter:

other: discharge

Basis:

mean

Remarks:

of 3 animals

Time point:

other: mean of 24, 48 and 72 hour evaluations

Score:

2.6

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Rinsed eyes

Irritant / corrosive response data:

The test material was found to cause moderate irritation.

Other effects:

No other effects reported.

Table 1 Calculation of Primary Eye Irritation Index

Time After Exposure	Mean Reaction Score
	Unrinsed Eyes			Rinsed Eyes
	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva
24 hrs	11.7	2.5	17.0	10.0	0.0	16.0
48 hrs	13.3	2.5	16.7	10.0	0.0	18.0
72 hrs	14.2	2.5	16.7	10.0	0.0	15.3
4 days	14.2	2.5	14.0	8.3	0.0	10.0
7 days	10.0	1.7	8.7	6.7	0.0	6.0
Subtotal	63.4	11.7	73.1	45.0	0.0	65.3
Total	148.2			110.3
PII	29.6			22.1

Table 2 Individual Irritation Scores at 24, 48, 72 hours and days 4 and 7

		Unrinsed							Rinsed
Observation Time	Observation	Male 1	Male 2	Male 3	Female 1	Female 2	Female 3	Average	Male 4	Male 5	Male 6	Average
24 hrs	Opacity	1	1	1	1	1	1	1.0	1	1	1	1.0
	Iris	1	1	1	0	0	0	0.5	0	0	0	0.0
	Conjunctivae	3	3	3	3	3	3	3.0	3	3	3	3.0
	Chemosis	3	3	3	3	3	2	2.8	3	2	3	2.7
48 hrs	Opacity	1	1	1	1	1	1	1.0	1	1	1	1.0
	Iris	1	1	1	0	0	0	0.5	0	0	0	0.0
	Conjunctivae	3	3	3	3	3	2	2.8	3	3	3	3.0
	Chemosis	3	3	3	3	3	1	2.7	3	3	3	3.0
72 hrs	Opacity	1	1	1	1	1	1	1.0	1	1	1	1.0
	Iris	1	1	1	0	0	0	0.5	0	0	0	0.0
	Conjunctivae	3	3	3	3	3	3	3.0	3	3	3	3.0
	Chemosis	3	3	3	2	3	3	2.8	3	2	2	2.3
4 days	Opacity	1	1	1	1	1	1	1.0	1	1	1	1.0
	Iris	1	0	1	0	1	0	0.5	0	0	0	0.0
	Conjunctivae	3	3	2	3	3	3	2.8	2	2	2	2.0
	Chemosis	2	2	2	2	2	3	2.2	2	2	2	2.0
7 days	Opacity	1	1	1	1	1	1	1.0	1	1	1	1.0
	Iris	1	0	1	0	0	0	0.3	0	0	0	0.0
	Conjunctivae	2	1	2	2	2	1	1.7	1	2	1	1.3
	Chemosis	2	1	2	2	2	1	1.7	1	1	1	1.0

Interpretation of results:

other: Classified as Category 2 according to EU criteria.

Conclusions:

Under the conditions of the test, the test material was shown to be irritating to the eyes. In accordance with EU criteria the test material is classified as Category 2.

Executive summary:

The eye irritancy potential of the test material was assessed in a GLP compliant in vivo study that was based upon the procedure outlined in the guidelines of the United States EPA, Paragraph 163.81-4 “Primary eye irritation study”, Federal Register, Vol. 43 No. 163, August 22, 1978.

Nine New Zealand White rabbits were exposed to the test material; the treated eye of three animals was rinsed with saline 30 seconds after exposure. The reactions to the test material were evaluated in accordance with the Draize scale (1977) after 24, 48 and 72 hours followed by further observations after 4 and 7 days. The test material was determined to cause moderate irritation.

Under the conditions of this study, the test material was shown to cause irritation to the eye which would require classification under regulation directive 1272/2008. The test material requires classification as "Category 2, H319: Causes serious eye irritation" with signal word "Warning".

Endpoint conclusion:

adverse effect observed (irritating)

Endpoint conclusion:

no study available

Skin Irritation

In the GLP compliant key study, the skin irritancy potential of the test material was assessed in vivo using a method based upon the procedure outlined in the guidelines of the United States EPA, Paragraph 163.81-5 “Primary dermal irritation study”, Federal Register, Vol. 43 No. 163, August 22, 1978.

Six New Zealand White rabbits were exposed to the test material in an occlusive fashion for 24 hours on both intact and abraded skin. The skin reactions were evaluated in accordance with the Draize scale at 24, 48 and 72 hours followed by observations after 4 and 7 days.

Under the conditions of this study, the test material produced marked irritation to the skin.

The study was performed to a good standard with a sufficient level of detail to assess the quality of the study. As such it was assigned a reliability score of 2 using the principles for assessing data quality as set out in Klimisch (1997) and considered suitable as an accurate reflection of the test material.

Eye Irritation

In the GLP compliant key study, the skin irritancy potential of the test material was assessed in vivo using a method based upon the procedure outlined in the guidelines of the United States EPA, Paragraph 163.81-4 “Primary eye irritation study”, Federal Register, Vol. 43 No. 163, August 22, 1978.

Nine New Zealand White rabbits were exposed to the test material; the treated eye of three animals was rinsed with saline 30 seconds after exposure. The reactions to the test material were evaluated in accordance with the Draize scale (1977) after 24, 48 and 72 hours followed by further observations after 4 and 7 days. The test material was determined to cause moderate irritation under the conditions of the study.

The study was performed to a good standard with a sufficient level of detail to assess the quality of the study. As such it was assigned a reliability score of 2 using the principles for assessing data quality as set out in Klimisch (1997) and considered suitable as an accurate reflection of the test material.

Justification for selection of skin irritation / corrosion endpoint:
Only one study available.

Justification for selection of eye irritation endpoint:
Only one study available.

Effects on skin irritation/corrosion: irritating

Effects on eye irritation: irritating

Skin Irritation

The skin irritation study indicates that the test material requires classification under regulation 1272/2008. The test material requires classification as “Category 2, H315: Causes skin irritation” with signal word “Warning”.

Eye Irritation

The eye irritation study indicates that the test material requires classification under regulation 1272/2008. The test material requires classification as “Category 2, H319: Causes serious eye irritation" with signal word " Warning".