Registration Dossier

- Skin irritation: not irritating in rabbits (one study acc. to OECD guideline 404 and EU method B.4, GLP), applied to the intact skin for 4 hours. At first only one animal was used and after 4 hours exposure to the test item, very slight erythema (hardly noticeable) (grade 1) was recorded one hour after removing the patch. Full reversibility was recorded 24 hours after application. 
- Eye irritation: irritating in rabbits (one study acc. to OECD guideline 405 and EU method B.5, GLP); undiluted test material applied to right eye of three animals. Moderate conjunctival redness and moderate to severe chemosis were observed in all three animals at 1 hour examination. Moderate conjunctival redness and well defined chemosis were observed in the three animals at 24 and 48 hours examination. In one animal, scattered or diffuse areas of opacity were recorded in the cornea, the details of the iris were clearly visible (grade 1) during the observation performed at one hour and up to 72 h following the administration. The cornea effects were fully reversible within 7 days. Two animals still had slight conjunctival effect on day 7 but all effects were fully reversed within 14 days.

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 27/7/2005 to 15/9/2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP 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

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Granja Cunícula San Bernardo s.L
- Age at study initiation: 10-11 weeks
- Weight at study initiation: 2.3-2.6 kg
- Housing: individually in stainless steel cages with a grille floor placed on racks.
- Diet (e.g. ad libitum): free access
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-25
- Humidity (%): 55-70
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

not required

Amount / concentration applied:

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

Duration of treatment / exposure:

At first only one animal was used, applying three patches sequently for 3 minutes, 1 and 4 hours. After each one of these exposure periods, the corresponding bandage and patch were removed. The remaining test item was remioved with water. The test was completed using two other animals, applying a dressing to each one for 4 hours. After this exposure period, the bandages and patches were removed. The remaining test item was removed wih water.

Observation period:

The response in the application area was assessed after 60 minutes, 24, 48 and 72 hours after the period of exposure.

Number of animals:

Three female

Details on study design:

TEST SITE
- Area of exposure: 6.2 cm^2

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with water

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3

Time point:

other: mean 24, 48, 72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal: #1, #2, #3

Time point:

other: mean 24, 48, 72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

mean

Remarks:

all animals

Time point:

other: 60 min

Score:

0.33

Reversibility:

fully reversible

Irritation parameter:

erythema score

Basis:

mean

Remarks:

all animals

Time point:

other: mean 24, 48 and 72 h

Score:

0

Irritation parameter:

edema score

Basis:

mean

Remarks:

all animals

Time point:

other: mean 24, 48, 72 h

Score:

0

Irritant / corrosive response data:

At first only one animal was used and after 4 hours exposure to the test item, very slight erythema (hardly noticeable) (grade 1) was recorded one hour after removing the patch.

Primary Dermal Irrtation Test
Nº animal (Exposure)	Response	60 min	24 h	48 h	72 h
2267 (3 minutes)	Erythema	0	0	0	0
2267 (3 minutes)	Edema	0	0	0	0
2267 (1Hour)	Erythema	0	0	0	0
2267 (1Hour)	Edema	0	0	0	0
2267 (4hours)	Erythema	1	0	0	0
2267 (4hours)	Edema	0	0	0	0

Primary Dermal Irrtation Test
Nº animal (Exposure)	Response	60 min	24 h	48 h	72 h
2267	Erythema	1	0	0	0
2267	Edema	0	0	0	0
2269	Erythema	0	0	0	0
2269	Edema	0	0	0	0
2271	Erythema	0	0	0	0
2271	Edema	0	0	0	0

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: CLP, EU GHS (Regulation (EC) No 1272/2008)

Conclusions:

The test item is considered to be NON IRRITANT based on the criteria set out in Directive 2001/59/EC, Annex VI, and CLP, EU GHS (Regulation (EC) No 1272/2008)

Executive summary:

In a primary dermal irritation study according to the OECD Guideline for the Testing of Chemicals 404 and Commission Directive 2004/73/EC Method B.4, 3 female, young adult New Zealand white rabbits were dermally exposed to 0.5 g of Phosphoric acid, mono- and di- C16-18 (even numbered) alkyl esters (100% a.i.) for 3 minutes, 1 and 4 hours to 6.2 cm² skin surface. Animals then were observed for 3 days.

No skin lesions were recorded after removing the patches applied for 3 minutes and 1 hour in the observation carried out 1 hour after removing the patches. After the 4 hour exposure to the test item, very slight erythema (hardly noticeable) (grade 1) was observed one hour after removing the patch. No skin lesions were observed in this animal during the observations made 24, 48 and 72 hours after removing the patches applied for 3 minutes, 1 hour or 4 hours. A semiocclusive patch with test tem was applied for 4 hours in the other two animals. No skin lesions were recorded in any of the animals during the observations made 1, 24, 48 and 72 hours after removing the patch. The means of the erythema and edema evaluations at 24, 48 and 72 hours for the three animals after a four-hour exposure were 0.00 (erythema) and 0.00 (edema).

In this study Phosphoric acid, mono- and di- C16-18 (even numbered) alkyl esters are not irritating to rabbit skin.

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:

From 29/12/2005 to 12/4/2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: 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

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Granja Cunícola San Bernardo, S.L
- Age at study initiation: 10-13 weeks
- Weight at study initiation: 2.4-3.0 Kg
- Housing: individually in stainless steel cages with a grille floor and placed on a rack
- Diet (e.g. ad libitum): free access
- Water (e.g. ad libitum): ad libitum
- Acclimation period: seven days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-23
- Humidity (%): 30-50
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

other: left eye as a Control

Amount / concentration applied:

The quantity of test item to be administered to each animal was 0.1 g.

Duration of treatment / exposure:

1 hour
The remainder of the test item was removed with physiological saline before the first observation.

Observation period (in vivo):

1, 24, 48, 72 hours, 7 and 14 days

Number of animals or in vitro replicates:

Three female

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

all animals

Time point:

other: 24, 48 and 72 hours

Score:

0.33

Reversibility:

fully reversible

Irritation parameter:

iris score

Basis:

mean

Remarks:

all animals

Time point:

other: 24, 48 and 72 hours

Score:

0

Reversibility:

fully reversible

Irritation parameter:

conjunctivae score

Remarks:

(Hyperaemia)

Basis:

mean

Remarks:

all animals

Time point:

other: 24, 48 and 72 hours

Score:

2.22

Reversibility:

fully reversible

Irritation parameter:

chemosis score

Remarks:

(Edema)

Basis:

mean

Remarks:

all animals

Time point:

other: 24, 48 and 72 hours

Score:

0.89

Reversibility:

fully reversible

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: mean 24, 48, 72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: mean 24, 48, 72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: mean 24, 48, 72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 14 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: mean 24, 48, 72 h

Score:

0.6

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: mean 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: mean 24, 48, 72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: mean 24, 48, 72 h

Score:

3

Max. score:

3

Reversibility:

fully reversible within: 14 days

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: mean 24, 48, 72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

other: mean 24, 48, 72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: mean 24, 48, 72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

other: mean 24, 48, 72 h

Score:

1.6

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: mean 24, 48, 72 h

Score:

0.6

Max. score:

4

Reversibility:

fully reversible within: 72 h

Body weight in Kg

Animal no.	Day of treatment	24h	48h	72h	7days	14days
2142	3,02	3,02	3,01	3,05	3,05	3,3
903	2,36	2,39	2,41	2,44	2,56	2,75
904	2,35	2,37	2,43	2,48	2,54	-

- : This animal was sacrified 7 days after the administration, as no ocular lesions were recorded

Primary Eye Irritation Test

Animal no.	Region of eye		1h	24h	48h	72h	7days	14days
2142	Cornea	Density	0	0	0	0	0	0
	Iris		0	0	0	0	0	0
	Conjunctivae	Hyperaemia Edema	2 4	2 2	2 0	2 0	1 0	0 0
903	Cornea	Density	1	1	1	1	0	0
	Iris		0	0	0	0	0	0
	Conjunctivae	Hyperaemia Edema	2 3	3 2	3 1	3 1	1 1	0 0
904	Cornea	Density	0	0	0	0	0	-
	Iris		0	0	0	0	0	-
	Conjunctivae	Hyperaemia Edema	2 2	2 1	2 1	1 0	0 0	-

-: This animal was sacrified 7 days after the administration, as no ocular lesions were recorded.

Interpretation of results:

irritating

Remarks:

Migrated information Category 2 Criteria used for interpretation of results: other: CLP, EU GHS (Regulation (EC) No 1272/2008)

Conclusions:

The alterations induced by the test item were evaluated to the OECD 405 and Method B.5. In view of the results obtained the test item is considered to be non irritant to the eyes based on the criteria set out in Directive 2001/59/EC, Annex VI, and no risk phrase is required. According to CLP, EU GHS (Regulation (EC) No 1272/2008) the test item has to be classified as irritating to eyes (Category 2).

Executive summary:

In a primary eye irritation study according to the OECD Guideline for the Testing of Chemicals 405 and Commission Directive 2004/73/EC Method B.5, 0.1 g of Phosphoric acid, mono- and di- C16-18 (even numbered) alkyl esters (100% a.i.) was instilled into the conjunctival sac of the right eye of three female, young adult New Zealand Whit rabbits for 1 hour. The remainder of the test item was removed with physiological saline before the first observation. Animals then were observed for 14 days.

The test item was administered initially to one animal. One hour after the application, diffuse crimson colouring (grade 2) was recorded in the conjunctiva, along with swelling with the lids more than half closed (grade 4). However, during the observations it was difficult to evaluate the hyperaemia due to the swelling of the conjunctiva. The test item was later applied to two more animals. Scattered or diffuse areas of opacity were recorded in the cornea, the details of the iris were clearly visible (grade 1) during the observation performed at one hour following the administration.

The mean values at 24, 48 and 72 hours following administration, for each type of lesion in the three treated animals were:

- Corneal opacity: 0.33

- Lesions in the iris: 0.00

- Hyperaemia: 2.22

- Edema: 0.89

In this study Phosphoric acid, mono- and di- C16-18 (even numbered) alkyl esters are irritating to the rabbit eye.

Endpoint conclusion:

adverse effect observed (irritating)

Endpoint conclusion:

no study available

Skin irritation

In a primary dermal irritation study according to the OECD Guideline for the Testing of Chemicals 404 and Commission Directive 2004/73/EC Method B.4, 3 female, young adult New Zealand white rabbits were dermally exposed to 0.5 g of Phosphoric acid, mono- and di- C16-18 (even numbered) alkyl esters (100% a.i.) for 3 minutes, 1 and 4 hours to 6.2 cm² skin surface. Animals then were observed for 3 days.

No skin lesions were recorded after removing the patches applied for 3 minutes and 1 hour in the observation carried out 1 hour after removing the patches. After the 4 hour exposure to the test item, very slight erythema (hardly noticeable) (grade 1) was observed one hour after removing the patch. No skin lesions were observed in this animal during the observations made 24, 48 and 72 hours after removing the patches applied for 3 minutes, 1 hour or 4 hours. A semiocclusive patch with test tem was applied for 4 hours in the other two animals. No skin lesions were recorded in any of the animals during the observations made 1, 24, 48 and 72 hours after removing the patch. The means of the erythema and edema evaluations at 24, 48 and 72 hours for the three animals after a four-hour exposure were 0.00 (erythema) and 0.00 (edema).

In this study Phosphoric acid, mono- and di- C16-18 (even numbered) alkyl esters are not irritating to rabbit skin.

Eye irritation

In a primary eye irritation study according to the OECD Guideline for the Testing of Chemicals 405 and Commission Directive 2004/73/EC Method B.5, 0.1 g of Phosphoric acid, mono- and di- C16-18 (even numbered) alkyl esters (100% a.i.) was instilled into the conjunctival sac of the right eye of three female, young adult New Zealand Whit rabbits for 1 hour. The remainder of the test item was removed with physiological saline before the first observation. Animals then were observed for 14 days.

The test item was administered initially to one animal. One hour after the application, diffuse crimson colouring (grade 2) was recorded in the conjunctiva, along with swelling with the lids more than half closed (grade 4). However, during the observations it was difficult to evaluate the hyperaemia due to the swelling of the conjunctiva. The test item was later applied to two more animals. Scattered or diffuse areas of opacity were recorded in the cornea, the details of the iris were clearly visible (grade 1) during the observation performed at one hour following the administration.

The mean values at 24, 48 and 72 hours following administration, for each type of lesion in the three treated animals were:

- Corneal opacity: 0.33

- Lesions in the iris: 0.00

- Hyperaemia: 2.22

- Edema: 0.89

In this study Phosphoric acid, mono- and di- C16-18 (even numbered) alkyl esters are irritating to the rabbit eye.

Respiratory irritation

No data on the respiratory irritation of Phosphoric acid, mono- and di- C16-18 (even numbered) alkyl esters are available.

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

Justification for selection of skin irritation / corrosion endpoint:
OECD & EC guideline study, no deviations, GLP

Justification for selection of eye irritation endpoint:
OECD & EC guideline study, no deviations, GLP

Effects on eye irritation: irritating

Skin irritation

From reliable, adequate and relevant data there is no indication on a skin irritating activity of Phosphoric acid, mono- and di- C16-18 (even numbered) alkyl esters, thus the classification criteria regarding skin irritation outlined in regulation (EC) 1272/2008 or the former European directive on classification and labelling 67/548/EEC are not met.

Eye irritation

Based on reliable, adequate and relevant data, Phosphoric acid, mono- and di- C16-18 (even numbered) alkyl esters have to be classified as irritating to eyes (Category 2) according to CLP, EU GHS (Regulation (EC) No 1272/2008) and is assigned the hazard statement H 319 and the signal word “Warning”. According to the criteria set out in directive 2011/59/EC, Annex VI, Phosphoric acid, mono- and di-C16-18-alkyl esters are considered to be non irritant to the eyes and no risk phrase is required.