Registration Dossier

Testing was carried out on the following endpoints using the following guidelines:

Skin irritation - OECD Guidelines for the Testing of Chemicals No. 404 “Acute Dermal Irritation/Corrosion” (adopted 24 April 2002).

Eye irritation - OECD Guidelines for the Testing of Chemicals No. 405 “Acute Eye Irritation/Corrosion” (adopted 24 April 2002).

The above studies have all been ranked reliability 1 according to the Klimisch system. This ranking was deemed appropriate because the studies were conducted to GLP and 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.

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

between 07 November 2006 and 10 November 2006.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 (incl. QA statement)

Remarks:

Date of inspection: 30/08/05; Date of signature: 19/01/07

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan UK Limited, Bicester, Oxon, UK.
- Age at study initiation: twelve to twenty weeks old.
- Weight at study initiation: 2.0 to 3.5 kg
- Housing: The animals were individually housed in suspended cages.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23°C
- Humidity (%): 30 to 70%
- Air changes (per hr): at least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

IN-LIFE DATES: From: Day 1 To: 72 hours after application.

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g
- Concentration (if solution): not applicable

VEHICLE
- Amount(s) applied (volume or weight with unit): test material moistened
- Concentration (if solution): not applicable
- Lot/batch no. (if required): not applicable
- Purity: not applicable

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: dorsal/flank area
- % coverage: 10%
- Type of wrap if used: cotton gauze patch secured in position with a strip of surgical adhesive tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): any residual test material removed by gentle swabbing with cotton wool soaked in distilled water.
- Time after start of exposure: 4 hours

SCORING SYSTEM: Draize scheme

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Other effects:

None

The individual scores for erythema/eschar and oedema are given in Table 1.

Very slight erythema was noted at all treated skin sites one hour after patch removal and at two treated skin sites at the 24-hour observation.

One treated skin site appeared normal at the 24-hour observation and the remaining two treated skin sites appeared normal at the 48-hour observation.

Table1 IndividualSkin Reactions

Skin Reaction	Observation Time	Individual Scores – Rabbit Number and Sex			Total
Skin Reaction	Observation Time	65078 Male	65079 Male	65080 Male	Total
Erythema/Eschar Formation	1 Hour	1	1	1	(3 )
	24 Hours	0	1	1	2
	48 Hours	0	0	0	( 0 )
	72 Hours	0	0	0	0
Oedema Formation	1 Hour	0	0	0	( 0 )
	24 Hours	0	0	0	0
	48 Hours	0	0	0	( 0 )
	72 Hours	0	0	0	0
Sum of 24 and 72-hour Readings (S): 2
Primary Irritation Index (S/6) : 2/6 = 0.3
Classification : MILD IRRITANT

( ) = Total values not used for calculation of primary irritation index

Interpretation of results:

GHS criteria not met

Conclusions:

The test material has a primary irritation index of 0.3. The average score after 24/48/72 hours for erythema was 0/0.33/0.33 for animal 1, 2 and 3. The average score after 24/48/72 hours for edema was 0/0/0 for animal 1, 2 and 3. Therefore the substance is not classified according to the EU CLP regulation (EC No 1272/2008) for skin irritation.

Executive summary:

Introduction.

The study was performed to assess the irritancy potential of the test material to the skin of the New Zealand White rabbit. The method was designed to meet the requirements of the following:

§ OECD Guidelines for the Testing of Chemicals No. 404 “Acute Dermal Irritation/Corrosion” (adopted)

§ Method B4 Acute Toxicity (Skin Irritation) of Commission Directive 2004/73/EC

Results.

A single 4-hour, semi-occluded application of the test material to the intact skin of three rabbits produced very slight erythema. One treated skin site appeared normal at the 24-hour observation and the remaining two treated skin sites appeared normal at the 48-hour observation.

Conclusion.

The test material has a primary irritation index of 0.3. The average score after 24/48/72 hours for erythema was 0/0.33/0.33 for animal 1, 2 and 3. The average score after 24/48/72 hours for edema was 0/0/0 for animal 1, 2 and 3. Therefore the substance is not classified according to the EU CLP regulation (EC No 1272/2008) for skin irritation.

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:

The study was performed between 13 November 2006 and 28 November 2006.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 (incl. QA statement)

Remarks:

Date of inspection: 30/08/05; Date of signature: 14/12/06

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Not stated in report.
- Age at study initiation: twelve to twenty weeks old.
- Weight at study initiation: 2.0 to 3.5 kg
- Housing: The animals were individually housed in suspended cages.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23°C
- Humidity (%): 30 to 70%
- Air changes (per hr): at least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

IN-LIFE DATES: From: Day 1 To: Day 7

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml of the test material, which was found to weigh approximately 30 mg (as measured by gently compacting the required volume into an adapted syringe)
- Concentration (if solution): not applicable

VEHICLE
- Amount(s) applied (volume or weight with unit): not applicable
- Concentration (if solution): not applicable
- Lot/batch no. (if required): not applicable
- Purity: not applicable

Duration of treatment / exposure:

7 days

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): not stated in report if washing was done.
- Time after start of exposure: not stated in report

SCORING SYSTEM: Kay and Calandra

TOOL USED TO ASSESS SCORE: Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

1.33

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

: Redness

Basis:

mean

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Individual and group mean scores for ocular irritation are given in Table1and Table2.

White residual test material was noted in all treated eyes at the 1-hour observation.

Scattered or diffuse corneal opacity was noted in one treated eye at the 24-hour observation, in all treated eyes at the 48-hour observation and persisted in one treated eye at the 72‑hour observation.

Iridial inflammation was noted in all treated eyes one hour after treatment and at the 24-hour observation and in one treated eye at the 48-hour observation.

Moderate conjunctival irritation was noted in all treated eyes one hour after treatment and at the 24, 48 and 72-hour observations.

All treated eyes appeared normal at the 7-day observation.

Table1 IndividualScores and Individual Total Scoresfor Ocular Irritation

Rabbit Number and Sex	65132 Male					65198 Male					65191 Male
Rabbit Number and Sex	IPR= 2					IPR = 2					IPR = 2
Time After Treatment	1 Hour	24 Hours	48 Hours	72 Hours	7 Days	1 Hour	24 Hours	48 Hours	72 Hours	7 Days	1 Hour	24 Hours	48 Hours	72 Hours	7 Days
CORNEA
E = Degree of Opacity	0	1	1	0	0	0	0	1	0	0	0	1	1	1	0
F = Area of Cornea Involved	0	2	1	0	0	0	0	1	0	0	0	2	1	1	0
Score (E x F) x 5	0	10	5	0	0	0	0	5	0	0	0	10	5	5	0
IRIS
D	1	1	0	0	0	1	1	1	0	0	1	1	0	0	0
Score (D x 5)	5	5	0	0	0	5	5	5	0	0	5	5	0	0	0
CONJUNCTIVAE
A = Redness	2	2	2	2	0	2	2	2	2	0	2	2	2	2	0
B = Chemosis	2	2	1	1	0	2	2	1	1	0	2	2	1	1	0
C = Discharge	2Re	2	1	1	0	2Re	2	1	1	0	2Re	2	1	1	0
Score (A + B + C) x 2	12	12	8	8	0	12	12	8	8	0	12	12	8	8	0
Total Score	17	27	13	8	0	17	17	18	8	0	17	27	13	13	0

IPR= Initial pain reaction

Re= White residual test material in the treated eye

Table2 Individual Total Scores and Group Mean Scoresfor Ocular Irritation

Rabbit Number and Sex	Individual Total Scores At:
Rabbit Number and Sex	1 Hour	24 Hours	48 Hours	72 Hours	7 Days
65132 Male	17	27	13	8	0
65198 Male	17	17	18	8	0
65191 Male	17	27	13	13	0
Group Total	51	71	44	29	0
Group Mean Score	17.0	23.7	14.7	9.7	0.0

IPR= Initial pain reaction

Re= White residual test material in the treated eye

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

The test material produced a maximum group mean score of 23.7 and was classified as a MODERATE IRRITANT (CLASS 5 ON A 1 TO 8 SCALE) to the rabbit eye according to a modified Kay and Calandra classification system.
The substance is classified as an eye irritant (Category 2: Causes serious eye irritation) under the EU CLP Regulation (EC) No 1272/2008.

Executive summary:

Introduction.

The study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method was designed to meet the requirements of the following:

§ OECD Guidelines for the Testing of Chemicals No. 405 “Acute Eye Irritation/Corrosion” (adopted)

§ Method B5 Acute Toxicity (Eye Irritation) of Commission Directive 2004/73/EC

Result.

A single application of the test material to the non-irrigated eye of three rabbits produced scattered or diffuse corneal opacity, iridial inflammation and moderate conjunctival irritation. All treated eyes appeared normal at the 7-day observation.

Conclusion.

The test material produced a maximum group mean score of 23.7 and was classified as a MODERATE IRRITANT (CLASS 5 ON A 1 TO 8 SCALE) to the rabbit eye according to a modified Kay and Calandra classification system.

The substance is classified as an eye irritant (Category 2: Causes serious eye irritation) under the EU CLP Regulation (EC) No 1272/2008.

Endpoint conclusion:

adverse effect observed (irritating)

Endpoint conclusion:

no study available

The studies listed above gave the following results:

Skin irritation.

A single 4-hour, semi-occluded application of the test material to the intact skin of three rabbits produced very slight erythema. One treated skin site appeared normal at the 24-hour observation and the remaining two treated skin sites appeared normal at the 48-hour observation.

Eye irritation.

A single application of the test material to the non-irrigated eye of three rabbits produced scattered or diffuse corneal opacity, iridial inflammation and moderate conjunctival irritation. All treated eyes appeared normal at the 7-day observation.

Effects on eye irritation: moderately irritating

The studies listed above triggered the following classification.

CLP Regulation (EC No 1272/2008).

The substance is classified as acutely irritating to the Eye: Category 2, H319 Causes serious eye damage. The substance is not classified as irritating to the skin.