Registration Dossier

Skin irritation: Not irritating, OECD 404, EU Method B.4, de Jouffery 1995.
Eye irritation: Not irritating, OECD 405, EU Method B.5, de Jouffery 1995.

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 June 1995 to 24 June 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guidelines and the study was conducted under GLP conditions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

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: 2.7 ± 0.1 kg
- Housing: Individually in polystyrene cages (35 x 55 x 32 cm or 48.2 x 58 x 36.5 cm).
- Diet: Pelleted diet ad libitum.
- Water: Filtered water ad libitum
- Acclimation period: At least 5 days.
- Animal selection: The day before treatment, the skin of ach animal was examined in order to use only animals without any signs of cutaneous irritation. Animals showing signs of cutaneous irritation, cutaneous defects or pre-existing dermal injury were not used.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3 °C
- Humidity (%): 30 – 70 %
- Photoperiod: 12 hour light/dark cycle.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 500 mg per animal as supplied.

Duration of treatment / exposure:

4 hours.

Observation period:

72 hours.

Number of animals:

3 male rabbits

Details on study design:

TEST SITE
- Area of exposure: 6 cm² on the right flank of each animal. The left flank (clipped) served as an untreated control.
- Type of wrap if used: The test material was prepared on a gauze pad moistened with 0.5 mL of distilled water, and held in place with an adhesive hypoallergenic aerated semi-occlusive dressing and a restraining bandage.

REMOVAL OF TEST SUBSTANCE
- Washing: Any residual test material was removed with gauze moistened with water.
- Time after start of exposure: 4 hours.

SCORING SYSTEM:
Cutaneous examinations were performed at approximately 1, 24, 48 and 72 hours after the removal of the dressing. Dermal irritation was evaluated in accordance with the Draize scale, see Table 1. Any other lesions were noted.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: Mean of observations at 24, 48 and 72 hours

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: Mean of observations at 24, 48 and 72 hours

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: Mean of observations at 24, 48 and 72 hours

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: Mean of observations at 24, 48 and 72 hours

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: Mean of observations at 24, 48 and 72 hours

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: Mean of observations at 24, 48 and 72 hours

Score:

0

Max. score:

4

Irritant / corrosive response data:

Very slight orange colouration of the test site which could not mask an erythema was observed on all animals during the study.
No skin reaction was noted in one animal.
A very slight or slight erythema (grade 1 or 2) was observed one hour after treatment in two of the animals. Erythema persisted for 24 hours in one rabbit only.
No oedema was noted at any time.

Table 2: Individual Dermal Reactions

Rabbit No.	Dermal Irritation	Scores				Mean Irritation Score
Rabbit No.	Dermal Irritation	1 hr	24 hrs	48 hrs	72 hrs	Mean Irritation Score
01	Erythema	1	0	0	0	0.0
	Oedema	0	0	0	0	0.0
	Other	C	C	C	*
02	Erythema	0	0	0	0	0.0
	Oedema	0	0	0	0	0.0
	Other	C	C	C	*
03	Erythema	2	1	0	0	0.3
	Oedema	0	0	0	0	0.0
	Other	C	C	C	C

Mean score calculated from readings taken at 24, 48 and 72 hours.

* = None

C = Orange colouration of the skin

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the test, the test material is considered to be a non-irritant since none of the animals displayed a classifiable reaction.

Executive summary:

The acute dermal irritation of the test material was determined in vivo in a study conducted under GLP conditions and in accordance with the standardised guidelines OECD 404 and EU method B.4.

Three male New Zealand White rabbits were exposed to 500 mg of the test material for 4 hours. Dermal irritation was assessed 1 hour after removal and then at 24, 48 and 72 hours after removal according to the Draize scale. Any other lesions were noted.

A very slight orange colouration of the test site which could not mask an erythema was observed on all animals during the study. No skin reaction was noted in one animal. A very slight or slight erythema (grade 1 or 2) was observed one hour after treatment in two of the animals. Erythema persisted for 24 hours in one rabbit only. No oedema was noted at any time.

Under the conditions of the test, the test material is considered to be a non-irritant since none of the animals displayed a reaction severe enough to trigger classification.

Endpoint conclusion:

no adverse effect observed (not irritating)

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 June 1995 to 02 July 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guidelines and the study was conducted under GLP conditions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.9 ± 0.2 kg
- Housing: Individually in polystyrene cages (35 x 55 x 32 cm or 48.2 x 58 x 36.5 cm).
- Diet: Pelleted diet ad libitum.
- Water: Filtered water ad libitum
- Acclimation period: At least 5 days.
- Animal selection: The day before treatment, the eyes of each animal were examined in order to use only animals without any signs of ocular irritation. Animals showing signs of ocular irritation, ocular defects or pre-existing corneal injury were not used.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3 °C
- Humidity (%): 30 – 70 %
- Photoperiod: 12 hour light/dark cycle.

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg per animal as supplied.

Duration of treatment / exposure:

A single exposure was performed.

Observation period (in vivo):

Animals were observed for 72 hours post application.

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

APPLICATION
- Method: The test material in its original form was placed into the conjunctival sac of the left eye of the 3 animals after gently pulling the lower lid away from the eyeball. The lower and upper lids were held together for about one second to avoid any loss of the test material. The right eye remained untreated as a control.

REMOVAL OF TEST SUBSTANCE
- Washing: Not performed.

SCORING SYSTEM: The eye was examined approximately 1 hour after administration and then at 24, 48 and 72 hours. The ocular reaction was scored according to the Draize scale, see Table 1. Any other lesions observed were noted.

TOOL USED TO ASSESS SCORE: Direct corneal examination was performed, if necessary with an Ultra Violet lamp. 1 or 2 drops of 0.5 % sodium fluorescein solution was used to determine the presence or absence of corneal opacification.

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

4

Irritation parameter:

other: redness

Basis:

animal #1

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

3

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

2

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

animal #1

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

4

Irritation parameter:

other: redness

Basis:

animal #2

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

3

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

2

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

animal #2

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

4

Irritation parameter:

other: redness

Basis:

animal #3

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

3

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

2

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

animal #3

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritant / corrosive response data:

An orange colouration of the conjunctiva was noted 1 hour after treatment only, in all animals; redness of conjunctiva was marked by this colouration.
In one animal a slight (grade 1) corneal opacity was noted at the 24 hour reading only.
No other ocular lesions were observed during the study.

Table 2: Ocular Response

Rabbit No.	Region of Eye	Description of Ocular Reactions	Scores				Mean Irritation Score
Rabbit No.	Region of Eye	Description of Ocular Reactions	1 hr	24 hrs	48 hrs	72 hrs	Mean Irritation Score
01	Conjunctivae	Chemosis	0	0	0	0	0.0
		Redness	C	0	0	0	0.0
		Discharge	0	0	0	0	0.0
	Iris		0	0	0	0	0.0
	Corneal Opacity	Intensity	0	0	0	0	0.0
	Corneal Opacity	Area	0	0	0	0	0.0
	Other		*	*	*	*
	Fluorescein		/	U	/	/
02	Conjunctivae	Chemosis	0	0	0	0	0.0
		Redness	C	0	0	0	0.0
		Discharge	0	0	0	0	0.0
	Iris		0	0	0	0	0.0
	Corneal Opacity	Intensity	0	0	0	0	0.0
	Corneal Opacity	Area	0	0	0	0	0.0
	Other		*	*	*	*
	Fluorescein		/	U	/	/
03	Conjunctivae	Chemosis	0	0	0	0	0.0
		Redness	C	0	0	0	0.0
		Discharge	0	0	0	0	0.0
	Iris		0	0	0	0	0.0
	Corneal Opacity	Intensity	0	1	0	0	0.3
	Corneal Opacity	Area	0	2	0	0	0.7
	Other		*	*	*	*
	Fluorescein		/	U	/	/

Mean score calculated from readings taken at 24, 48 and 72 hours.

* = None

C = Scoring masked by orange colouration

U = Fluorescein used

/ = Fluorescein not used

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the test, the test material was determined to be non-irritating, since a classifiable reaction was not observed in any of the treated rabbits.

Executive summary:

The acute eye irritation of the test material was determined in vivo in a study conducted under GLP conditions and in accordance with the standardised guidelines OECD 405 and EU method B.5.

Three male New Zealand White rabbits were exposed to 100 mg of the test material in a single ocular application. Ocular irritation was assessed 1 hour after application and then at 24, 48 and 72 hours, according to the Draize scale. Any other lesions were noted.

An orange colouration of the conjunctiva was noted 1 hour after treatment only, in all animals; redness of conjunctiva was obscured by this colouration. In one animal a slight (grade 1) corneal opacity was noted at the 24 hour reading only. No other ocular lesions were observed during the study.

Under the conditions of the test, the test material is considered to be a non-irritant since none of the animals displayed a reaction strong enough to trigger classification.

Endpoint conclusion:

no adverse effect observed (not irritating)

Endpoint conclusion:

no study available

SKIN IRRITATION

The acute dermal irritation of the test material was determined in a key study by de Jouffery (1995). The study was performed in vivo under GLP conditions and in accordance with the standardised guidelines OECD 404 and EU method B.4.

Three male New Zealand White rabbits were exposed to 500 mg of the test material for 4 hours. Dermal irritation was assessed 1 hour after removal and then at 24, 48 and 72 hours after removal according to the Draize scale. Any other lesions were noted.

A very slight orange colouration of the test site which could not mask an erythema was observed on all animals during the study. No skin reaction was noted in one animal. A very slight or slight erythema (grade 1 or 2) was observed one hour after treatment in two of the animals. Erythema persisted for 24 hours in one rabbit only. No oedema was noted at any time.

Under the conditions of the test, the test material is considered to be a non-irritant since none of the animals displayed a reaction sever enough to trigger classification.

EYE IRRITATION

The acute eye irritation of the test material was determined in a key study by de Jouffery (1995). The study was performed in vivo in under GLP conditions and in accordance with the standardised guidelines OECD 405 and EU method B.5.

Three male New Zealand White rabbits were exposed to 100 mg of the test material in a single ocular application. Ocular irritation was assessed 1 hour after application and then at 24, 48 and 72 hours, according to the Draize scale. Any other lesions were noted.

An orange colouration of the conjunctiva was noted 1 hour after treatment only, in all animals; redness of conjunctiva was obscured by this colouration. In one animal a slight (grade 1) corneal opacity was noted at the 24 hour reading only. No other ocular lesions were observed during the study.

Under the conditions of the test, the test material is considered to be a non-irritant since none of the animals displayed a reaction strong enough to trigger classification.

Justification for selection of skin irritation / corrosion endpoint:
This is the only available study that addresses acute skin irritation of the test material. The study was performed under GLP conditions to standardised guidelines. The report is conclusive and reported to a sufficient standard for classification. In accordance with the principles for assessing data quality defined by Klimisch et al (1997), this study was assigned a reliability score of 1.

Justification for selection of eye irritation endpoint:
This is the only available study that addresses acute eye irritation of the test material. The study was performed under GLP conditions to standardised guidelines. The report is conclusive and reported to a sufficient standard for classification. In accordance with the principles for assessing data quality defined by Klimisch et al (1997), this study was assigned a reliability score of 1.

SKIN IRRITATION

In accordance with criteria for classification as defined in Annex I, Regulation 1272/2008, the test material does not require classification as a skin irritant.

EYE IRRITATION

In accordance with the criteria for classification as defined in Annex I, Regulation 1272/2008, the test material does not require classification as an eye irritant.