Registration Dossier

Skin irritation

- Not irritating to skin; OECD TG 404 (RL1, GLP), in vivo, rabbit, 4 h, semiocclusive: edema & erythema scores 0/0/0

Eye irritation:

irritating to the eyes (Category 2); OECD TG 405 (RL1, GLP), in vivo, rabbit: cornea scores 1/1/1, iris scores 0.3/0.7/0.3, conjunctivae scores 2.7/3.0/2.7, chemosis scores 1.7/1.3/1.7

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007-07-05 to 2007-07-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

(2004)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

(2002)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: at least 6 weeks
- Weight at study initiation: at least 1.0 kg
- Housing: Individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56x44x37.5 cm).
- Diet (e.g. ad libitum): Pelleted diet for rabbits (K-H from SSNlFF® Spezialdiaten GmbH, Soest, Germany) approximately 100 grams per day. Hay (TecniLab-BMI BV, Someren, The Netherlands) was provided at least three times a week.
- Water (e.g. ad libitum): ad libitum tap water
- Acclimation period: at least 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.3 - 22.6°C
- Humidity (%): 45 - 68%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

other: adjacent areas of the untreated skin of each animal served as controls

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g

Duration of treatment / exposure:

4 hours

Observation period:

72 hours after removal of the test substance

Number of animals:

3 males

Details on study design:

TEST SITE
The test substance was applied to the skin of one flank, using a metalline patch of 2x3 cm. The patch was mounted on Micropore tape, which was wrapped around the abdomen and secured with Coban elastic bandage.

REMOVAL OF TEST SUBSTANCE
Four hours after the application, the dressing was removed and the skin cleaned of residual test substance using tap water.

SCORING SYSTEM:
as stipulated in OECD TG 404

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

There was no evidence of a corrosive effect on the skin. No skin irritation was caused by 4 hours exposure to Amphopropionate C8.

Other effects:

No staining of the treated skin by the test substance was observed and no test substance remnants were seen. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Interpretation of results:

GHS criteria not met

Conclusions:

In this study Amphopropionate C8 was not irritating/corrosive to the skin.
Mean erythema scores from observations at 24, 48 and 72 h after patch removal were 0 for 3/3 animals; mean edema scores were 0 for 3/3 animals.

Executive summary:

In a primary dermal irritation study according to OECD guideline 404 (2002) and EU method B.4 (2004) 3 young adult male New Zealand White rabbits were dermally exposed to 0.5 g of Amphopropionate C8 (97% a.i.) for 4 hours to 6 cm² body surface area. Animals then were observed for 72 hours. Irritation was scored according to the Draize scale as stipulated in the test guideline.

No skin irritation/corrosion was caused by 4 hours exposure to Amphopropionate C8. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred. Mean erythema and mean edema scores from observations at 24, 48 and 72 h after patch removal were 0 for 3/3 animals.

Based on these results, Amphopropionate C8 does not have to be classified and has no obligatory labelling requirement for skin irritation according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).

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:

2007-07-09 to 2007-09-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age at study initiation: >/= 9 weeks
- Weight at study initiation: 2.165 - 2.757 kg
- Housing: individually
- Diet (e.g. ad libitum): pelleted diet for rabbits (K-H from SSNIFF Spezialdiäten)
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: at least 5 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0±3 (actual range: 21.0-22.4°C)
- Humidity (%): 30-70 (actual range: 42-78%)
- Air changes (per hr): approx. 15
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL of the test item was instilled in the conjunctival sac of one eye

Duration of treatment / exposure:

24 h

Observation period (in vivo):

14 d

Number of animals or in vitro replicates:

3 (started by treatment of a single rabbit (sentinel); the two other animals were treated 6 weeks later, after considering the degree of eye irritation observed in the first animal)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no; at the 24 h observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage.

SCORING SYSTEM: according to OECD TG 405 (Draize scale)

TOOL USED TO ASSESS SCORE: hand-slit lamp / fluorescein

Irritation parameter:

cornea opacity score

Basis:

animal: #1, 2, 3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 d

Irritation parameter:

iris score

Basis:

animal: #1, 3

Time point:

24/48/72 h

Score:

0.3

Max. score:

2

Reversibility:

fully reversible within: 72 h

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.7

Max. score:

2

Reversibility:

fully reversible within: 7 d

Irritation parameter:

conjunctivae score

Basis:

animal: #1, 3

Time point:

24/48/72 h

Score:

2.7

Max. score:

3

Reversibility:

fully reversible within: 14 d

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

fully reversible within: 14 d

Irritation parameter:

chemosis score

Basis:

animal: #1, 3

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 7 d

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 14 d

Irritant / corrosive response data:

Instillation of 0.1 mL of the test item into one eye of each of three rabbits resulted in effects on the cornea, iris and conjunctivae. The corneal injury consisted of opacity (maximum grade 1) and epithelial damage (maximum 75 or 50% of the corneal area). The corneal injury had resolved within 7 days. Iridial irritation grade 1 was observed and had resolved within 48 hours, 72 hours or 7 days. The irritation of the conjunctivae consisted of redness, chemosis and discharge and had completely resolved within 14 days.
There was no evidence of ocular corrosion.
No staining of (peri) ocular tissues by the test substance was observed and no test substance remnants were seen.

Other effects:

No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Irritant/corrosive response data for each animal at each observation time up to removal of each animal from the test

Score at time point	Cornea	Iris	Conjunctivae	Chemosis
Score at time point	Max. score: 4	Max. score: 2	Max. score: 3	Max. score: 4
1 h	1/1/1	1/1/1	2/2/2	2/3/3
24 h	1/1/1	0/1/1	2/3/2	2/2/3
48 h	1/1/1	1/0/0	3/3/3	2/1/1
72 h	1/1/1	0/1/0	3/3/3	1/1/1
Average 24h, 48h, 72h	1/1/1	0.3/0.7/0.3	2.7/3.0/2.7	1.7/1.3/1.7
7 d	0/0/0	0/0/0	1/2/1	0/1/0
14 d	0/0/0	0/0/0	0/0/0	0/0/0

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

In this study, of Amphopropionate C8 (97% a.i.) is irritating to the eye (Category 2).

Executive summary:

In a primary eye irritation study according to OECD Guideline 405 (2002) and EU method B.5 (2004) 0.1 mL of Amphopropionate C8 (97% a.i.) was instilled into the conjunctival sac of one eye of 3 male young adult New Zealand White rabbits. At the 24 h observation, a solution of 2% fluorescein in water was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. Animals then were observed for 14 days. Irritation was scored by the method of Draize.

Instillation of 0.1 mL of the test item into one eye of each of three rabbits resulted in effects on the cornea, iris and conjunctivae. The corneal injury consisted of opacity (maximum grade 1) and epithelial damage (maximum 75 or 50% of the corneal area). The corneal injury had resolved within 7 days. Iridial irritation grade 1 was observed and had resolved within 48 hours, 72 hours or 7 days. The irritation of the conjunctivae consisted of redness, chemosis and discharge and had completely resolved within 14 days. There was no evidence of ocular corrosion.

In this study, Amphopropionate C8 (97% a.i.) is irritating to the eye (Category 2) based on conjunctival redness scores >/= 2.

Endpoint conclusion:

adverse effect observed (irritating)

Endpoint conclusion:

no study available

Skin irritation

In a primary dermal irritation study according to OECD guideline 404 (2002) and EU method B.4 (2004) 3 young adult male New Zealand White rabbits were dermally exposed to 0.5 g of Amphopropionate C8 (97% a.i.) for 4 hours to 6 cm² body surface area. Animals then were observed for 72 hours. Irritation was scored according to the Draize scale as stipulated in the test guideline.

No skin irritation/corrosion was caused by 4 hours exposure to Amphopropionate C8. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred. Mean erythema and mean edema scores from observations at 24, 48 and 72 h after patch removal were 0 for 3/3 animals.

Based on these results, Amphopropionate C8 does not have to be classified and has no obligatory labelling requirement for skin irritation according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).

Eye irritation

In a primary eye irritation study according to OECD Guideline 405 (2002) and EU method B.5 (2004) 0.1 mL of Amphopropionate C8 (97% a.i.) was instilled into the conjunctival sac of one eye of 3 male young adult New Zealand White rabbits. At the 24 h observation, a solution of 2% fluorescein in water was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. Animals then were observed for 14 days. Irritation was scored by the method of Draize.

Instillation of 0.1 mL of the test item into one eye of each of three rabbits resulted in effects on the cornea, iris and conjunctivae. The corneal injury consisted of opacity (maximum grade 1) and epithelial damage (maximum 75 or 50% of the corneal area). The corneal injury had resolved within 7 days. Iridial irritation grade 1 was observed and had resolved within 48 hours, 72 hours or 7 days. The irritation of the conjunctivae consisted of redness, chemosis and discharge and had completely resolved within 14 days. There was no evidence of ocular corrosion.

In this study, Amphopropionate C8 (97% a.i.) is irritating to the eye (Category 2) based on conjunctival redness scores >/= 2.

There are no data gaps for the endpoint irritation/corrosion. No human data are available. However, there is no reason to believe that these results from rabbit would not be applicable to humans.

Skin irritation

Based on reliable, adequate and relevant data, Amphopropionate C8 does not need to be classified for skin irritation according to regulation (EC) 1272/2008.

Eye irritation

Based on reliable, adequate and relevant data, Amphopropionate C8 is classified as Category 2 (Irritating to eyes) according to regulation (EC) 1272/2008 and labelled with H319: Causes serious eye irritation and the signal word “Warning”.