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 15 January and 05 February 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study conducted in compliance with OECD Guideline No. 404 without any deviation.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

24 April 2002.

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Directive n° 2004/73/EC.

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

Inspected on 2007-01-11 / Signed on 2007-02-21.

Specific details on test material used for the study:

- The test item was considered at 100% for the study.
- Date received: 17 December 2007

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: 2.42 - 3.05 kg
- Housing: Each animal was kept in an individual box installed in conventional air conditioned animal husbanding.
- Diet: foodstuff, ad libitum
- Water: tap-water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20-22 °C
- Humidity: 37- 64 %
- Air changes: no data
- Photoperiod: 12 hours light/12 hours dark.

IN-LIFE DATES: From 15 January to 05 February 2008.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes

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 one 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: After removal of the patch, the treated area was rinsed with distilled water.
- Time after start of exposure: 4 h

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:

1.7

Max. score:

4

Reversibility:

fully reversible within: 6 days

Remarks on result:

probability of mild irritation

Irritation parameter:

edema score

Basis:

animal #1

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 weak irritation

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 6 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:

1.3

Max. score:

4

Reversibility:

fully reversible within: 6 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:

1.7

Max. score:

4

Reversibility:

fully reversible within: 6 days

Remarks on result:

probability of mild irritation

Irritation parameter:

edema 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 weak irritation

Irritant / corrosive response data:

- A well defined erythema associated with a slight oedema was noted, 24 hours after the test item application, on the treated area.
- The oedematous reactions were totally reversible between D3 and D6 and the erythematous reactions were totally reversible at D6.
- On the cutaneous structure, a slight dryness was noted on the treated area, from D2 in one animal and from D3 in the two others animals. The skin recovered a normal aspect between D13 and D14.

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)	A8715/Male	A8738/Male	A8739/Male
Erythema/Eschar formation	1 h	1	2	2
	24 h	2	2	2
	48 h	2	2	2
	72 h	1	2	1
	Day 6	0	0	0
	Total (24, 48 and 72 hours)	5	6	5
	Mean (24, 48 and 72 hours)	1.7	2	1.7

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)	A8715/Male	A8738/Male	A8739/Male
Oedema formation	1 h	0	1	1
	24 h	2	2	2
	48 h	1	1	1
	72 h	0	1	0
	Day 6	0	0	0
	Total (24, 48 and 72 hours)	3	4	3
	Mean (24, 48 and 72 hours)	1	1.3	1

Note:

A8715: slight dryness at D2, D3, D6 to D10 &D13

A8738: slight dryness at D3, D6 to D10

A8739: slight dryness at D3, D6 to D10 &D13

Interpretation of results:

Category 3 (mild irritant) based on GHS criteria

Conclusions:

Under the test conditions, the test substance is not classified as irritating to skin according to the criteria of the Annex I 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 well defined erythema associated with a slight oedema was noted, 24 hours after the test item application, on the treated area. The oedematous reactions were totally reversible between D3 and D6 and the erythematous reactions were totally reversible at D6. On the cutaneous structure, a slight dryness was noted on the treated area, from D2 in one animal and from D3 in the two others animals. The skin recovered a normal aspect between D13 and D14.

The mean scores for each animal within 3 scoring times (24, 48 and 72 h) were 1.7 / 2.0 / 1.7 for erythema and 1.0 / 1.3 / 1.0 for oedema.

Under the test conditions, test substance is not classified as irritating to skin according to the criteria of the Annex I 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 January 2008.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study conducted in compliance with OECD Guideline No. 404 without any deviation.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

24 April 2002.

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Directive 2004/73/EC

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

Inspected on 2007-01-11 / Signed on 2007-02-21.

Specific details on test material used for the study:

- The test item was considered at 100% for the study.
- Date received: 17 December 2007

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: 2.39 - 2.70 kg
- Housing: the animal was kept in an individual box installed in conventional air conditioned animal husbanding.
- Diet: foodstuff, ad libitum
- Water: tap-water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20-22 °C
- Humidity: 40-57 %
- Air changes: no data
- Photoperiod: 12 hours light/12 hours dark.

IN-LIFE DATES: from 21 to 25 January 2008.

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 following treatment, according to a numerical evaluation.

Number of animals or in vitro replicates:

3 females

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 responses produced in the first treated animal at D1, 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:

0.3

Max. score:

4

Reversibility:

fully reversible within: 2 days

Remarks on result:

probability of weak 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:

0.3

Max. score:

3

Reversibility:

fully reversible within: 2 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

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

no indication of 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

Max. score:

3

Reversibility:

other: not applicable

Remarks on result:

no indication of 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:

0.3

Max. score:

4

Reversibility:

not fully reversible within: 2 days

Remarks on result:

probability of weak 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 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 D1 and D2, associated with a slight chemosis, noted only 1 hour after the test item instillation;
- at the corneal level: a slight opacity, only registered 24 hours after the test item instillation in two animals.

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
A8756 Female	1 h	0	0	1	1
	24 h	1	0	1	0
	48 h	0	0	0	0
	72 h	0	0	0	0
	Total (24, 48 and 72 h)	1	0	1	0
	Mean (24, 48 and 72 h)	0.3	0	0.3	0

Rabbit Number and Sex	Time After Treatment	Corneal Opacity	Iris lesion	Conjunctival Redness	Conjunctival Chemosis
A8754 Female	1 h	0	0	1	1
	24 h	0	0	0	0
	48 h	0	0	0	0
	72 h	0	0	0	0
	Total (24, 48 and 72 h)	0	0	0	0
	Mean (24, 48 and 72 h)	0	0	0	0

Rabbit Number and Sex	Time After Treatment	Corneal Opacity	Iris lesion	Conjunctival Redness	Conjunctival Chemosis
A8755 Female	1 h	0	0	1	1
	24 h	1	0	1	0
	48 h	0	0	0	0
	72 h	0	0	0	0
	Total (24, 48 and 72 h)	1	0	2	0
	Mean (24, 48 and 72 h)	0.3	0.0	0.3	0.0

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the test substance is not classified as irritating to eyes according to the criteria of annex I of 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 female 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 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.

The ocular reactions observed during the study have been slight and totally reversible in the three animals. A slight redness, noted 1 hour after the test item instillation and totally reversible between D1 and D2, associated with a slight chemosis, noted only 1 hour after the test item instillation. At the corneal level, a slight opacity was registered 24 hours after the test item instillation in two animals.

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0.3, 0.0, 0.3 for cornea score; 0.0, 0.0, 0.0 for iris score; 0.3, 0.0, 0.3 for conjunctivae score and 0.0, 0.0, 0.0 for chemosis score.

Under the test conditions, the test substance is not classified as irritating to eyes according to the criteria of annex I of 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, 2008). 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 well defined erythema associated with a slight oedema was noted, 24 hours after the test item application, on the treated area. The oedematous reactions were totally reversible between D3 and D6 and the erythematous reactions were totally reversible at D6. On the cutaneous structure, a slight dryness was noted on the treated area, from D2 in one animal and from D3 in the two others animals. The skin recovered a normal aspect between D13 and D14.

The mean scores for each animal within 3 scoring times (24, 48 and 72 h) were 1.7 / 2.0 / 1.7 for erythema and 1.0 / 1.3 / 1.0 for oedema.

Eye irritation:

A key study was identified (Phycher, 2008). 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 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.

The ocular reactions observed during the study have been slight and totally reversible in the three animals. A slight redness, noted 1 hour after the test item instillation and totally reversible between D1 and D2, associated with a slight chemosis, noted only 1 hour after the test item instillation. At the corneal level, a slight opacity was registered 24 hours after the test item instillation in two animals.

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0.3, 0.0, 0.3 for cornea score; 0.0, 0.0, 0.0 for iris score; 0.3, 0.0, 0.3 for conjunctivae score and 0.0, 0.0, 0.0 for chemosis score.

Harmonized classification:

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

Self classification:

Skin irritation:

Based on the available data:

- no additional 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 additional self-classification is proposed regarding eye irritation according to the CLP and to the GHS.

Respiratory irritation:

No data was available regarding respiratory irritation.