Registration Dossier

Skin irritation: not irritating (OECD 404, GLP, K, Rel.2)

Eye irritation: not irritating (OECD 405, GLP, K, Rel.2)

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 14 May 2013 and 04 June 2013.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

GLP study conducted in compliance with OECD Guideline No. 404 without any deviation. The substance is adequately identified, but details on composition are missing. Therefore validation applies with restrictions.

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:

30 May 2008

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

Inspected on 2012-04-23&24 / Signed on 2012-07-18.

Specific details on test material used for the study:

Date received: 22 April 2013

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: 2.33 - 3.03 kg.
- Housing: Each animal was kept in an individual box installed in conventional air conditioned animal husbanding.
- Diet: foodstuff (SDS – C15), ad libitum
- Water: tap-water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 17-23 °C
- Humidity: 30-70 %
- Air changes: 13 changes/h
- Photoperiod: 12 h dark / 12 h light

IN-LIFE DATES: From 14 May to 04 June, 2013.

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 females

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: after the removal of the patch, the treated area was rinsed with distilled water.

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

Max. score:

4

Reversibility:

fully reversible within: 7 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:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

probability of mild 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: 14 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:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

probability of mild irritation

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 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:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days.

Remarks on result:

probability of mild irritation

Irritant / corrosive response data:

- A well defined erythema was noted on the treated area of the animals, 24 hours after the patch removal. This reaction was totally reversible between days 7 and 14.
- A slight oedema was noted on the treated area of the animals, 24 hours after the patch removal. This reaction was totally reversible between days 7 and 14.
- On the cutaneous structure, dryness was noted from day 7 in all animals and was remaining on day 14.

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)	A3063/female	A3099/female	A3100/female
Erythema/Eschar formation	1 h	1	2	1
	24 h	2	2	2
	48 h	2	2	2
	72 h	2	2	2
	Day 7	0	1	1
	Day 14	0	0	0
	Total (24, 48 and 72 hours)	6	6	6
	Mean (24, 48 and 72 hours)	2.0	2.0	2.0

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)	A3063/female	A3099/female	A3100/female
Oedema formation	1 h	1	2	2
	24 h	2	2	2
	48 h	2	2	2
	72 h	2	2	2
	Day 7	0	1	0
	Day 14	0	0	0
	Total (24, 48 and 72 hours)	6	6	6
	Mean (24, 48 and 72 hours)	2.0	2.0	2.0

Note:

Dryness was noted from day 7 in all animals and was remaining on day 14.

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 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 female 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 was noted on the treated area of the animals, 24 hours after the patch removal. This reaction was totally reversible between days 7 and 14. A slight oedema was noted on the treated area of the animals, 24 hours after the patch removal. This reaction was totally reversible between days 7 and 14.

On the cutaneous structure, dryness was noted from day 7 in all animals and was remaining on day 14.

The mean scores for each animal within 3 scoring times (24, 48 and 72 h) were 2.0 / 2.0 / 2.0 for erythema and 2.0 / 2.0 / 2.0 for oedema. These reactions are totally reversible between days 7 and 14.

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 20 May and 13 June, 2013.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

GLP study conducted in compliance with OECD Guideline No. 405 without any deviation. The substance is adequately identified, but details on composition are missing. Therefore validation applies with restrictions.

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)

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

Inspected on 2012-04-23&24 / Signed on 2012- 07-18

Specific details on test material used for the study:

Date received: 22 April 2013

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: 2.58 - 3.44 kg
- Housing: Each animal was kept in an individual box installed in conventional air conditioned animal husbanding.
- Diet: foodstuff (SAFE – 112), ad libitum
- Water: tap-water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 17-23 °C
- Humidity: 30-70 %
- Air changes: 13 changes/h
- Photoperiod: 12 h dark / 12 h light

IN-LIFE DATES: from 20 May to 13 June, 2013.

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 & from D7 to D21 in order to determine the reversible character of the lesions observed (in case of persistent reactions), 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:

0.7

Max. score:

4

Reversibility:

fully reversible within: 21 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:

1.3

Max. score:

3

Reversibility:

fully reversible within: 14 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:

0.7

Max. score:

4

Reversibility:

fully reversible within: 14 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:

0.3

Max. score:

3

Reversibility:

fully reversible within: 2 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:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

no indication of 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:
- at the conjunctivae level: a slight to moderate redness noted 1 hour after the test item instillation and totally reversible between days 2 and 14, associated with a slight to moderate chemosis noted 1 hour after the test item instillation and totally reversible between days 1 and 14;
- at the corneal level: a slight corneal opacity, noted 48 hours after the test item instillation in one animal, and totally reversible on day 21.

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
A3041 Male	1 h	0	0	1	1
	24 h	0	0	1	0
	48 h	1	0	2	1
	72 h	1	0	1	1
	Day 7	1	0	1	1
	Day 14	1	0	0	0
	Day 21	0	0	0	0
	Total (24, 48 and 72 h)	2	0	4	2
	Mean (24, 48 and 72 h)	0.7	0	1.3	0.7

Rabbit Number and Sex	Time After Treatment	Corneal Opacity	Iris lesion	Conjunctival Redness	Conjunctival Chemosis
A3141 Male	1 h	0	0	2	2
	24 h	0	0	1	0
	48 h	0	0	0	0
	72 h	0	0	0	0
	Total (24, 48 and 72 h)	0	0	1	0
	Mean(24, 48 and 72 h)	0	0	0.3	0

Rabbit Number and Sex	Time After Treatment	Corneal Opacity	Iris lesion	Conjunctival Redness	Conjunctival Chemosis
A3142 Male	1 h	0	0	2	2
	24 h	0	0	1	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	0.3	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 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 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 from D7 to D21 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 to moderate redness noted 1 hour after the test item instillation and totally reversible between days 2 and 14, associated with a slight to moderate chemosis noted 1 hour after the test item instillation and totally reversible between days 1 and 14. A slight corneal opacity, noted 48 hours after the test item instillation in one animal, and totally reversible on day 21.

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0.7, 0.0, 0.0 for cornea score; 0.0, 0.0, 0.0 for iris score; 1.3, 0.3, 0.3 for conjunctivae score and 0.7, 0.0, 0.0 for chemosis score. These effects are reversible between days 2 and 21.

Under the test conditions, the test material is not classified as irritating to eyes according to the 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, 2013). In this dermal irritation study performed according to the OECD Guideline No. 404, and in compliance with GLP, 3 female 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 was noted on the treated area of the animals, 24 hours after the patch removal. This reaction was totally reversible between days 7 and 14. A slight oedema was noted on the treated area of the animals, 24 hours after the patch removal. This reaction was totally reversible between days 7 and 14.

On the cutaneous structure, dryness was noted from day 7 in all animals and was remaining on day 14.

The mean scores for each animal within 3 scoring times (24, 48 and 72 h) were 2.0 / 2.0 / 2.0 for erythema and 2.0 / 2.0 / 2.0 for oedema. These reactions are totally reversible between days 7 and 14.

Eye irritation:

A key study was identified (Phycher, 2013). 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 male 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 from D7 to D21 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 to moderate redness noted 1 hour after the test item instillation and totally reversible between days 2 and 14, associated with a slight to moderate chemosis noted 1 hour after the test item instillation and totally reversible between days 1 and 14. A slight corneal opacity, noted 48 hours after the test item instillation in one animal, and totally reversible on day 21.

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0.7, 0.0, 0.0 for cornea score; 0.0, 0.0, 0.0 for iris score; 1.3, 0.3, 0.3 for conjunctivae score and 0.7, 0.0, 0.0 for chemosis score. These effects are reversible between days 2 and 21.

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 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.