Registration Dossier

Not skin irritant.

Eye Irrit. 2, H319

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

according to guideline

Guideline:

other: Royal Decree 363/1995, of 10 March

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

other: Royal Decree 1078/93 of 2 July

Deviations:

not specified

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: authorized breeder.
- Weight at study initiation: 2001, 2453, 2607 g
- Housing: single housed in a stainless steel cage, brand Tecniplast.
- Diet: daily dose of 150 g special diet for rabbits experimental provided by an authorized provider.
- Water: filtered tap water, ad libitum.
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 2°C
- Humidity: 55 ± 25 %
- Air changes: 15 ACH with air filtered 5 µm.
- Photoperiod: 12 hour cycle dark/light

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

water

Remarks:

moistened

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g

Duration of treatment / exposure:

4 hours

Observation period:

24, 48, 72 hours, 7, 14 and 21 days

Number of animals:

3 male rabbits

Details on study design:

TEST SITE
- Area of exposure: 20 cm x 13 cm
- Coverage: 6 cm^2
- Type of wrap if used: Gauze type dressings (Melolin) or equivalent, Sticking plaster, hypoallergenic adhesive bandage (Fixomull Seretch) or equivalent.

REMOVAL OF TEST SUBSTANCE
- Washing: yes; after the exposure period the substance is gently removed with water or suitable solvent.

EVALUATION OF SKIN REACTIONS
Eschar formation
No erythema...................................................0
Very slight erythema (barely perceptible)......1
Well-defined erythema....................................2
Moderate to severe erythema.........................3
Severe erythema to slight eschar formation...4

Oedema formation
No oedema..............................................................................................................0
Very slight oedema (barely perceptible).................................................................1
Slight oedema (edges well-defined)........................................................................2
Moderate oedema (raised ca. 1 mm).......................................................................3
Severe oedema (raised more than 1 mm; extending beyond area of exposure)....4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: shaved skin

Irritation parameter:

erythema score

Basis:

other: animal 2 and 3

Time point:

24/48/72 h

Score:

ca. 0.33

Max. score:

4

Reversibility:

fully reversible within: 2 days

Remarks on result:

other: shaved skin

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

other: shaved skin

Other effects:

In the three animals was observed a brown skin coloured into the application site. This effect persists after 21 days in the rabbit no. 1 and disappears after 14 days in the rabbits no. 2 and no. 3.

Interpretation of results:

other: Not classified, according to the Regulation EC 1272/2008 (CLP)

Conclusions:

The substance was tested following method described in the Spain Royal Decree 363/1995 and Royal Decree1078/93. Under the experimental conditions the substance does not show skin irritation properties.

Executive summary:

The substance has been tested for skin irritation according to the procedure inside the Spain Royal Decree 363/1995 and Royal Decree1078/93.

Three male rabbits are tested with 0.5 g of test item to the shaved skin after 4 hours of patch treatment. The scores of each animal at the reading times of 24, 48 and 72 hours was used in calculating the respective mean values for each type of lesion.

The values of erythema and oedema, calculated as mean individual scores at 24, 48 and 72 hours, are below the trigger values for classification (2.3), therefore the substance is not classified under the CLP Regulation (EC n. 1272/2008).

Endpoint conclusion:

no adverse effect observed (not irritating)

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

The procedure of the test did not include the rinsing of the eye. Due to the intrinsic irritating nature of dusty materials, the experiments could have overestimated the irritating potential of the substance related to its chemical/toxicological properties.

Qualifier:

according to guideline

Guideline:

other: Real Decreto 363/1995 of 10 March

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

other: Real Decreto 1078/93 of 2 July

Deviations:

not specified

GLP compliance:

no

Species:

rabbit

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Housing: Tecniplast inox cages.
- Diet: 150 g per day standard diet.
- Water: tap water; ad libitum.
- Acclimation period: 7 days.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 2°C
- Humidity: 55 ± 25 %
- Air changes: 15 ACH
- Photoperiod: 12 hours dark

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

0.1 g fine powder.

Observation period (in vivo):

at 1, 24, 48 and 72 hours and at 7, 14 and 21 day after application.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no

SCORING SYSTEM:
Cornea
Opacity: degree of density
No ulceration or opacity........................................................................................... 0
Scattered or diffuse areas of opacity .................................................................... 1
Easily discernible translucent area, details of iris slightly obscured........................2
Nacrous area, no details of iris visible, size of pupil barely discernible.................. 3
Opaque cornea, iris not discernible through the opacity..........................................4

Iris
Normal...................................................................................................................... 0
Markedly deepened rugae, congestión, swelling ................................................... 1
No reaction to light, hemorrhage, gross destructlon (any or all of these) ...............2

Conjunctivae - redness
Blood vessels normal ...............................................................................................0
Some blood vessels definitely hyperemic (injected) ................................................1
Diffuse, crimson color, individual vessels not easily discernible............................. 2
Diffuse beefy red......................................................................................................3

Conjunctivae - chemosis
No swelling............................................................................................................... 0
Any swelling above normal (includes nictitating membranes)..................................1
Obvious swelling with partial eversión of lids.......................................................... 2
Swelling with lids about half closed .........................................................................3
Swelling with lids more than half-closed...................................................................4

TOOL USED TO ASSESS SCORE: hand-slit lamp.

Irritation parameter:

cornea opacity score

Basis:

other: 3/3 animals

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible

Irritation parameter:

iris score

Basis:

other: 3/3 animals

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible

Irritation parameter:

conjunctivae score

Basis:

other: 2/3 animals

Time point:

24/48/72 h

Score:

>= 2

Reversibility:

not fully reversible within: 21 days

Remarks:

At rabbit #2 and #3, on 21th day, was given a conjunctival redness score of 1.

Irritation parameter:

chemosis score

Basis:

other: 2/3 animals

Time point:

24/48/72 h

Score:

>= 2

Reversibility:

fully reversible

Mean values for cornea, iris, conjunctiva at 24, 48 and 72 hours.

	Rabbit #1	Rabbit #2	Rabbit #3	Mean
Corneal opacity	1	1	1	1
Iritis	0	0.67	1	0.55
Conjunctival redness	2	2.67	3	2.55
Conjunctival oedema	0.67	3	3.67	2.44

	Individual findings - Rabbit #1
	1 h	24 hrs	48 hrs	72 hrs	7 days	14 days	21 days
Corneal opacity	-	1	1	1	0	0	0
Iritis	1	0	0	0	0	0	0
Conjunctival redness	3	2	2	2	2	1	0
Conjunctival oedema	2	1	1	0	0	0	0

	Individual findings - Rabbit #2
	1 h	24 hrs	48 hrs	72 hrs	7 days	14 days	21 days
Corneal opacity	0	1	1	1	0	0	0
Iritis	0	1	1	0	0	0	0
Conjunctival redness	0	3	3	2	2	2	1
Conjunctival oedema	3	4	3	2	1	0	0

	Individual findings - Rabbit #2
	1 h	24 hrs	48 hrs	72 hrs	7 days	14 days	21 days
Corneal opacity	0	1	1	1	1	0	0
Iritis	0	1	1	1	1	0	0
Conjunctival redness	0	3	3	3	2	2	1
Conjunctival oedema	3	4	4	3	2	1	0

Interpretation of results:

other: Classified as Eye Irrit. 2, according to the Regulation EC 1272/2008 (CLP).

Conclusions:

The substance was tested for eye irritation following a mehod described in the Real Decreto 363/1995.Under the experimental conditions the substance is classified as Eye Irrit. 2, H319.

Executive summary:

The test was performed according to the Real Decreto 363/1995.

The eye irritation potential was investigated by instillation of the equivalent of 0.1 g/animal of the test substance. The scores of each animal at the reading times of 24, 48 and 72 hours was used in calculating the respective mean values for each type of lesion, as stated by the Regulation EC 1272/2008 (CLP).

Findings

Scores for corneal opacity and iritis were lower than the threshold value of 1 for 3/3 rabbits.

Scores for conjunctival redness and oedema were higher than the threshold value of 2 in 2/3 rabbits.

Every type of lesion was fully reversible within 21 days except the conjunctival redness response, which was identified in 2/3 rabbits on the 21th day of the test (graded as 1 according to the used scoring system).

From the present results the substance should be classified as corrosive to the eyes bacause the produced effects were not fully reversible within 21 days. However, the following considerations should be underlined.

Since the study did foresee the rising of eyes after the application of the test item in its solid form, the results of the test can be considered as an overestimation of the true irritating effects strictly related to the substance chemical/toxicological properties. Indeed, due to the intrinsic irritating nature of dusty materials, this overestimation is true especially for the conjunctival redness response and the fact that this lesion was not fully reversible within 21 days can be attributed to the lack of eye rinsing. Moreover, from the individual scoring of each animal, it can be observed a progressive reduction of this lesion, suggesting that the effect will most likely not be persistent.

Endpoint conclusion:

adverse effect observed (irritating)

Endpoint conclusion:

no study available

Skin irritation assessment

The assessment was based on a study made in 2000 according to the procedure inside the Spain Royal Decree 363/1995 and Royal Decree 1078/93.

Three male rabbits were tested with 0.5 g of test item to the shaved skin after 4 hours of patch treatment. Under the conditions of the present experiment the substance was found to cause no irritation when applied to intact rabbit skin.

Eye irritation assessment

The assessment was based on a study made in 2001 according to the procedure inside the Spain Royal Decree 363/1995 and Royal Decree 1078/93.

The eye irritation potential was investigated by instillation of the equivalent of 0.1 g/animal of the test substance. The scores of each animal at the reading times of 24, 48 and 72 hours was used in calculating the respective mean values for each type of lesion, as stated by the Regulation EC 1272/2008 (CLP).

Experimental findings:

Scores for corneal opacity and iritis were lower than the threshold value of 1 for 3/3 rabbits.

Scores for conjunctival redness and oedema were higher than the threshold value of 2 in 2/3 rabbits.

Every type of lesion was fully reversible within 21 days except the conjunctival redness response, which was identified in 2/3 rabbits on the 21th day of the test (graded as 1 according to the used scoring system).

From the present results the substance should be classified as corrosive to the eyes bacause the produced effects were not fully reversible within 21 days. However, the following considerations should be underlined.

Since the study did foresee the rising of eyes after the application of the test item in its solid form, the results of the test can be considered as an overestimation of the true irritating effects strictly related to the substance chemical/toxicological properties. Indeed, due to the intrinsic irritating nature of dusty materials, this overestimation is true especially for the conjunctival redness response and the fact that this lesion was not fully reversible within 21 days can be attributed to the lack of eye rinsing. Moreover, from the individual scoring of each animal, it can be observed a progressive reduction of this lesion, suggesting that the effect will most likely not be persistent.

According to the CLP Regulation (EC n. 1272/2008) the following classifications apply:

Skin irritant cat2:

1) Mean value of ≥ 2,3 - ≤ 4,0 for erythema/ eschar or for oedema in at least 2 of 3 tested animals from gradings at 24, 48 and 72 hours after patch removal or, if reactions are delayed, from grades on 3 consecutive days after the onset of skin reactions; or

2)Inflammation that persists to the end of the observation period normally 14 days in at least 2 animals, particularly taking into account alopecia (limited area), hyperkeratosis, hyperplasia, and scaling; o

3) In some cases where there is pronounced variability of response among animals, with very definite positive effects related to chemical exposure in a single animal but less than the criteria above.

Eye irritant Cat 2:

at least in 2 of 3 tested animals, a positive response of:

1) corneal opacity ≥ 1 and/or

2) 2) iritis ≥ 1, and/or

3) conjunctival redness ≥ 2 and/or

4) conjunctival oedema (chemosis) ≥ 2

5) calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material, and which fully reverses within an observation period of 21 days

Regarding the substance

Scores for corneal opacity and iritis were lower than the threshold value of 1 for 3/3 rabbits.

Scores for conjunctival redness and oedema were higher than the threshold value of 2 in 2/3 rabbits.

Every type of lesion was fully reversible within 21 days except the conjunctival redness response, which was identified in 2/3 rabbits on the 21th day of the test (graded as 1 according to the used scoring system).

From the present results the substance should be classified as corrosive to the eyes bacause the produced effects were not fully reversible within 21 days. However, the following considerations should be underlined.

Since the study did not foresee the rinsing of eyes after the application of the test item in its solid form, the results of the test can be considered as an overestimation of the true irritating effects strictly related to the substance chemical/toxicological properties. Indeed, due to the intrinsic irritating nature of dusty materials, this overestimation is true especially for the conjunctival redness response and the fact that this lesion was not fully reversible within 21 days can be attributed to the lack of eye rinsing. Moreover, from the individual scoring of each animal, it can be observed a progressive reduction of this lesion, suggesting that the effect will most likely not be persistent.

For the above mentioned reason, the substance does not meet the criteria for classification as Eye Damage 1. Instead, a classification as Eye Irrit 2, under the Regulation EC 1272/2008 (CLP), is warranted.