Registration Dossier

Skin irritation: not irritating (OECD 404, GLP, K, Rel.1)

Eye irritation: not irritating (OECD 405, GLP, K, Rel.1)

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 08 and 20 November, 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study conducted in compliance with OECD Guideline No. 404 with deviations not affecting the reliability of the study: details on feeding and environmental conditions not reported

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

24 April 2002.

Deviations:

yes

Remarks:

details on feeding and environmental conditions not reported.

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Directive n° 2004/73/EC.

Deviations:

yes

Remarks:

details on feeding and environmental conditions not reported.

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

Inspected on 2004-07-01 / Signed on 2004-09-13.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage de Gérome (Quartier Labaste – F40260 Linxe).
- Weight at study initiation: 2.74 - 3.46 kg.
- Housing: Each animal was kept in an individual box installed in conventional air conditioned animal husbanding.
- Diet: no data
- Water: no data
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 17-22 °C
- Humidity: 30-58 %
- Air changes: no data
- Photoperiod: no data

IN-LIFE DATES: From 08 to 20 November, 2005.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): Test item was applied as supplied.

Duration of treatment / exposure:

4 hours

Observation period:

Skin reactions were appreciated 1, 24, 48 and 72 hours after removal of the patch and observations were continued from Days 4 to 14 (in case of persistent reactions were observed).

Number of animals:

3 males

Details on study design:

PRETEST
Initially, a single animal was treated. After consideration of the cutaneous responses produced in the first treated animal, two additional animals were treated.

TEST SITE
- Area of exposure: Undamaged skin area of the right flank of each animal
- Type of wrap if used: Patch was secured in position with a strip of surgical adhesive tape.

REMOVAL OF TEST SUBSTANCE
- Washing: no data.

OBSERVATION TIME POINTS
1, 24, 48 and 72 hours after removal of the patch and observations were continued from Days 4 to 14 (in case of persistent reactions were observed).

SCORING SYSTEM:
- Method of calculation: Skin irritant reaction was scored as per Draize scale according to OECD guideline 404

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

2.3

Max. score:

4

Reversibility:

fully reversible within: 4 days

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 2 days

Remarks on result:

probability of weak irritation

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 4 days

Remarks on result:

probability of mild irritation

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 2 days

Remarks on result:

probability of weak irritation

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 3 days

Remarks on result:

probability of weak irritation

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable.

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

- A slight to important erythema in the 3 animals, associated with a slight oedema in two animals, was noted 24 hours after the test item application.
- The oedematous reactions were totally reversible the 3rd day of the test and the erythematous reactions were totally reversible between the 4th and the 5th day of the test.
- On the cutaneous structure, the skin recovered a normal aspect, between the 5th and the 12th day of the test.

Other effects:

None

Table 7.3.1/1: Individual and mean skin reactions/Erythema - Eschar formation following 4 hour exposure

Skin reaction	Observation time (following patch removal)	Individual Scores – Rabbit Number and Sex
Skin reaction	Observation time (following patch removal)	A6977/male	A6981/male	A6982/male
Erythema/Eschar formation	1 h	2	2	1
	24 h	3	2	1
	48 h	2	2	1
	72 h	2	1	0
	Day 4	0	0	0
	Total (24, 48 and 72 hours)	7	5	2
	Mean (24, 48 and 72 hours)	2.3	1.7	0.7

Table 7.3.1/2: Individual and mean skin reactions/Oedema formation following 4 hour exposure

Skin reaction	Observation time (following patch removal)	Individual Scores – Rabbit Number and Sex
Skin reaction	Observation time (following patch removal)	A6977/male	A6981/male	A6982/male
Oedema formation	1 h	1	0	0
	24 h	1	1	0
	48 h	0	0	0
	72 h	0	0	0
	Day 7	0	0	0
	Total (24, 48 and 72 hours)	1	1	0
	Mean (24, 48 and 72 hours)	0.3	0.3	0

Note:

A6977: slight dryness from D2 to D3 and from D9 to D10, dryness from D4 to D8

A6981: slight dryness at D2, dryness from D3 to D4

A6982: slight dryness from D2 to D3

Interpretation of results:

Category 3 (mild irritant) based on GHS criteria

Conclusions:

Under the test conditions, test substance is not classified as irritating to skin according to the criteria of the Regulation EC No. 1272/2008 (CLP) and classified in Category 3 (mild irritant) according to the GHS.

Executive summary:

In a dermal irritation study performed according to the OECD Guideline No. 404, and in compliance with GLP, 0.5 mL of undiluted test item was applied on an undamaged skin area of the flank of 3 male New Zealand White rabbits. On the other flank an untreated area was served as the control. Test sites were covered with a semi-occlusive dressing for 4 h. Skin irritation was assessed and scored according to the Draize scale at 1, 24, 48 and 72 hours after removal of the patch and observations were continued from Days 4 to 14. Initially, a single animal was treated. After consideration of the cutaneous responses produced in the first treated animal, two additional animals were treated.

A slight to important erythema in the 3 animals, associated with a slight oedema in two animals was noted 24 hours after the test item application. The oedematous reactions were totally reversible the 3rd day of the test and the erythematous reactions were totally reversible between the 4th and the 5th day of the test. On the cutaneous structure, the skin recovered a normal aspect, between the 5th and the 12th day of the test.

The mean scores for each animal within 3 scoring times (24, 48 and 72 h) were 2.3 / 1.7 / 0.7 for erythema and 0.3 / 0.3 / 0.0 for oedema.

Under the test conditions, test substance is not classified as irritating to skin according to the criteria of the Regulation EC No. 1272/2008 (CLP) and classified in Category 3 (mild irritant) according to the GHS.

This study is considered as acceptable and satisfies the requirement for skin irritation endpoint.

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:

Between 21 and 25 November 2005.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study conducted in compliance with OECD Guideline No. 405 with deviations not affecting the reliability of the study: details on feeding and environmental conditions not reported.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

24 April 2002.

Deviations:

yes

Remarks:

details on feeding and environmental conditions not reported.

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Directive n° 2004/73/EC.

Deviations:

yes

Remarks:

details on feeding and environmental conditions not reported.

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

Inspected on 2004-07-01 / Signed on 2004-09-13.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage de Gérome (Quartier Labaste – F40260 Linxe).
- Weight at study initiation: 2.25 - 3.24 kg
- Housing: Each animal was kept in an individual box installed in conventional air conditioned animal husbanding.
- Diet: no data
- Water: no data
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 18-23 °C
- Humidity: 26-43 %. A relative humidity of 26% was registered instead of 30% (minimal limit) as planned in the experimental protocol. This deviation did not, in any case, influence the development and the results of the study.
- Air changes: no data.
- Photoperiod: no data.

IN-LIFE DATES: from 21 to 25 November, 2005.

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL

- Concentration (if solution): Test item was instilled, as supplied

Duration of treatment / exposure:

No washing was done

Observation period (in vivo):

Ocular examinations were performed on both right and left eyes 1, 24, 48 and 72 h & Day 4 following treatment, according to a numerical evaluation.

Number of animals or in vitro replicates:

3 males

Details on study design:

TREATMENT
- 0.1 mL of the test item was instilled, as supplied, into the conjunctival sac of one eye. The other eye remained untreated serving as control.
- Initially, a single animal was treated. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

REMOVAL OF TEST SUBSTANCE
- Washing: No

SCORING SYSTEM: According to OECD guideline 405

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #1

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 4 days

Remarks on result:

probability of weak irritation

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 2 days

Remarks on result:

probability of weak irritation

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable.

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable.

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 3 days.

Remarks on result:

probability of weak irritation

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable.

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 3 days

Remarks on result:

probability of mild irritation

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable.

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 2 days.

Remarks on result:

probability of weak irritation

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable.

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The ocular reactions observed during the study have been slight to moderate, and totally reversible in the three animals:
- at the conjunctivae level: a slight redness, noted 1 hour after the test item instillation and totally reversible between the 3rd and the 5th day of the test, associated with a slight to moderate chemosis, noted 1 hour after the test item instillation and totally reversible between the 2nd and the 3rd day of the test ;
- at the corneal level: a moderate opacity, registered 24 hours after the test item instillation, in only one animal, and totally reversible the 4th day of the test.

Other effects:

None

Table 7.3.2/1: Individual and Mean Scores for Cornea, Iris and Conjunctivae

Rabbit Number and Sex	Time After Treatment	Corneal Opacity	Iris lesion	Conjunctival Redness	Conjunctival Chemosis
A 6986 Male	1 h	0	0	1	1
	24 h	0	0	1	1
	48 h	0	0	1	0
	72 h	0	0	1	0
	Day 4	0	0	0	0
	Total (24, 48 and 72 h)	0	0	3	1
	Mean (24, 48 and 72 h)	0	0	1	0.3

Rabbit Number and Sex	Time After Treatment	Corneal Opacity	Iris lesion	Conjunctival Redness	Conjunctival Chemosis
A7020 Male	1 h	0	0	1	1
	24 h	0	0	1	0
	48 h	0	0	1	0
	72 h	0	0	0	0
	Day 4	0	0	0	0
	Total (24, 48 and 72 h)	0	0	2	0
	Mean (24, 48 and 72 h)	0	0	0.7	0

Rabbit Number and Sex	Time After Treatment	Corneal Opacity	Iris lesion	Conjunctival Redness	Conjunctival Chemosis
A 7021 Male	1 h	0	0	1	2
	24 h	2	0	1	0
	48 h	1	0	0	0
	72 h	0	0	0	0
	Day 4	0	0	0	0
	Total (24, 48 and 72 h)	3	0	0	0
	Mean (24, 48 and 72 h)	1	0	0.3	0

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the test material is not classified as irritating to eyes according to the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

Executive summary:

In an eye irritation study conducted according to OECD 405 guideline and in compliance with GLP, 3 New Zealand White male rabbits were exposed to 0.1 mL of test item in one eye while the contralateral eye remained untreated and served as control. The eyes were examined and the changes were observed at 1, 24, 48 and 72 h and day 4 following treatment and graded according to the Draize method. Initially, a single animal was treated. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

A slight redness noted 1 hour after the test item instillation and totally reversible between the 3rd and the 5th day of the test, associated with a slight to moderate chemosis, noted 1 hour after the test item instillation and totally reversible between the 2nd and the 3rd day of the test. At the corneal level, a moderate opacity, registered 24 hours after the test item instillation, in only one animal, and totally reversible the 4th day of the test.

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0.0, 0.0, 1.0 for cornea score; 0.0, 0.0, 0.0 for iris score; 1.0, 0.7, 0.3 for conjunctivae score and 0.3, 0.0, 0.0 for chemosis score. These effects are reversible between days 2 and 4.

Under the test conditions, the test material is not classified as irritating to eyes according to the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for eye irritation endpoint.

Endpoint conclusion:

no adverse effect observed (not irritating)

Endpoint conclusion:

no study available

Skin irritation:

A key study was identified (Phycher, 2005). In this dermal irritation study performed according to the OECD Guideline No. 404, and in compliance with GLP, 3 male New Zealand White rabbits were exposed for 4 hours to a semi-occluded application of 0.5 mL of undiluted test material to the intact skin. Initially, a single animal was treated. After consideration of the cutaneous responses produced in the first treated animal, two additional animals were treated.

A slight to important erythema in the 3 animals, associated with a slight oedema in two animals was noted 24 hours after the test item application. The oedematous reactions were totally reversible the 3rd day of the test and the erythematous reactions were totally reversible between the 4th and the 5th day of the test. On the cutaneous structure, the skin recovered a normal aspect, between the 5th and the 12th day of the test.

The mean scores for each animal within 3 scoring times (24, 48 and 72 h) were 2.3 / 1.7 / 0.7 for erythema and 0.3 / 0.3 / 0.0 for oedema.

Eye irritation:

A key study was identified (Phycher, 2005). In this eye irritation study performed according to the OECD guideline 405, and in compliance with GLP, 0.1 mL of undiluted test material was instilled into one eye of 3 female New Zealand White rabbits. The other eye remained untreated and served as control. The eyes were examined and the changes were observed at 1, 24, 48 and 72 h and Day 4 following treatment and graded according to the Draize method. Initially, a single animal was treated. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

A slight redness noted 1 hour after the test item instillation and totally reversible between the 3rd and the 5th day of the test, associated with a slight to moderate chemosis, noted 1 hour after the test item instillation and totally reversible between the 2nd and the 3rd day of the test. At the corneal level, a moderate opacity, registered 24 hours after the test item instillation, in only one animal, and totally reversible the 4th day of the test.

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0.0, 0.0, 1.0 for cornea score; 0.0, 0.0, 0.0 for iris score; 1.0, 0.7, 0.3 for conjunctivae score and 0.3, 0.0, 0.0 for chemosis score. These effects are reversible between days 2 and 4.

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008 (CLP).

Self classification:

Skin irritation:

Based on the available data:

- no self-classification is proposed regarding skin irritation according to the CLP

- the substance is classified in "Category 3 (mild irritant)" according to the GHS.

Eye irritation:

Based on the available data, no self-classification is proposed regarding eye irritation according to the CLP and to the GHS.

Respiratory irritation:

No data was available regarding respiratory irritation.