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Administrative data

Description of key information

Skin Irritation, York (1985)

Under the conditions of this study the test material was considered to be moderately irritating.

Eye Irritation RE 85.09 (1985)

Under the conditions of this study the test material was considered to be a moderate eye irritant.

Eye Irritation RE 85.05 (1985)

Under the conditions of this study 10 μL of 50 % test material was considered to produce moderate/severe irritation, 10 μL of undiluted test material was considered to produce slight irritation.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 February 1985 to 09 May 1985
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
no guideline followed
Principles of method if other than guideline:
0.5 mL of test material was applied to the clipped dorsum of rabbits for 4 hours under a semi-occlusive patch. The irritation reactions were assessed immediately after removal of the patch and after 24, 48 and 72 hours.
GLP compliance:
not specified
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: commercial supplier
- Age at study initiation: 9 - 12 weeks old
- Housing: rabbits were housed individually in suspended cages with wire mesh floors
- Diet: pelleted rabbit diet with vitamin and mineral supplements ad libitum
- Water: ad libitum
- Animals are used for testing when the hair is in a non-growth (telogen) phase of the hair cycle.

Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 0.5 mL
- Concentration: undiluted
Duration of treatment / exposure:
4 hours
Observation period:
72 hours
Number of animals:
8
Details on study design:
TEST SITE
- Three to 4 days prior to testing the backs and flanks of the rabbits were clipped to identify those which were in telogen and which had no skin blemishes. Of these rabbits, 8 were selected for the test. The hair is given a final clip approximately 2 hours prior to treatment.
- Type of wrap if used: Each patch consisted of a 25 x 25 mm, 16-ply gauze pad backed by a 20 x 30 mm strips of thin polythene and stuck to a 25 x 75 mm strip of zinc oxide plaster.
- When all patches had been applied to a rabbit it was immobilised in a canvas body sleeve and placed in a quiet place for the 4 hour application period, at the end, of which the animals were removed from the body sleeves and the corners of each treatment site marked.

REMOVAL OF TEST SUBSTANCE
- The patches were removed and excess substance wiped from the skin with a damp tissue.

OBSERVATION TIME POINTS
- The skin treatment sites were assessed for irritation immediately after removal of the patches and at 24, 48 and 72 hours after treatment.

SCORING SYSTEM:
- Each animal was scored blind. Reactions were assessed for erythema, oedema, cracking and scaling on a 9-point scale ranging from very slight (a) to severe (h). Any other features of the responses were described and recorded as a fraction of treated area.
- Reaction grades:
- = no reaction: normal skin
a = marginal (very slight)
b = slight
c = fairly distinct:
d = quite distinct
e = becoming well developed
f = well developed
g = becoming severe
h = severe
- At the end of the test the irritation reaction produced by each test material at 24 and 72 hours after treatment was assigned a single overall irritation score which was recorded along with the irritation scores. This allowed for comparisons of responses within and between experimental groups. The effects of the test and control materials were compared along with any important or unusual features of the test.

- Overall irritation scores
0 = NORMAL SKIN
1 = MARGINAL: Slight erythema and oedema (grade a) at 24 hours but no effects observed at 72 hours.
2 = SLIGHT: Slight erythema and oedema (grades up to bbbb) at 24 and 72 hours.
3 = SLIGHT/MODERATE: Slight/moderate erythema and oedema (grades ca-cc) at 24 hours. The reaction generally reduces to slight at 72 hours but sometimes the slight/moderate reaction may be maintained.
4 = MODERATE: Moderate erythema and oedema (grades c and d) with slight cracking and scaling at 24 hours with the erythema and oedema being maintained at least at slight/moderate level with cracking and scaling developing at 72 hours.
5 = STRONG: As for 4 above or greater erythema and oedema with evidence of up to 1/4 of the site affected by necrosis.
6 = SEVERE: Moat of site affected by suspected necrosis.
7 = EXTREME: Deep necrosis over whole site identified by depressed, hard scab/tissue and surrounding inflammatory reaction.
Irritation parameter:
overall irritation score
Basis:
animal #1
Time point:
24/48/72 h
Score:
4
Max. score:
7
Reversibility:
not specified
Irritation parameter:
overall irritation score
Basis:
animal #2
Time point:
24/48/72 h
Score:
4
Max. score:
7
Reversibility:
not specified
Irritation parameter:
overall irritation score
Basis:
animal #3
Time point:
24/48/72 h
Score:
5
Max. score:
7
Reversibility:
not specified
Irritation parameter:
overall irritation score
Basis:
animal #4
Time point:
24/48/72 h
Score:
4
Max. score:
7
Reversibility:
not specified
Irritation parameter:
overall irritation score
Basis:
animal #5
Time point:
24/48/72 h
Score:
2
Max. score:
7
Reversibility:
not specified
Irritation parameter:
overall irritation score
Basis:
animal #6
Time point:
24/48/72 h
Score:
3
Max. score:
7
Reversibility:
not specified
Irritation parameter:
overall irritation score
Basis:
animal: #7
Time point:
24/48/72 h
Score:
4
Max. score:
7
Reversibility:
not specified
Irritation parameter:
overall irritation score
Basis:
animal: #8
Time point:
24/48/72 h
Score:
6
Max. score:
7
Reversibility:
not specified
Irritant / corrosive response data:
The test material produced distinct erythema and oedema at 4 hours which intensified slightly to show variable but generally slight/moderate effects at 72 hours. Two animals developed extensive areas of brown tissue.

Table 1: Daily Assessment of Irritation Reactions

Rabbit No.

4 Hours

24 Hours

48 Hours

72 Hours

er

oe

cr

sc

er

oe

cr

sc

er

oe

cr

sc

er

oe

cr

sc

1

d

c

-

-

d

b

a

a

c

b

b

a

c

b

b

b

2

d

b

-

-

d

c

-

-

c

b

-

-

b

a

-

-

3

b

a

-

-

d

c

-

-*

c

c

-

-*

e

b

a

a*

4

d

c

-

-

d

c

-

-

d

b

a

a

c

b

b

a

5

c

b

-

-

b

a

-

-

b

a

-

-

b

a

-

-

6

d

b

-

-

c

b

-

-

b

a

a

a

b

a

a

a

7

b

a

-

-

d

c

-

-

c

b

a

a

c

b

b

a

8

c

b

-

-

d

b

-

-*

d

b

-

-*

d

c

a

a*

er = erythema

oe = oedema

cr = cracking

sc = scaling

* Macroscopic reactions:

24 Hours:

Rabbit 1: 10 x 10 mm green/brown tissue

Rabbit 8: Whole area pale brown tissue

48 Hours:

Rabbit 1: 10 x 10 mm green/brown tissue

Rabbit 8: Whole area pale brown tissue

72 Hours:

Rabbit 1: 10 x 10 mm green/brown tissue

Rabbit 8: Whole area pale brown tissue

Conclusions:
Under the conditions of this study the test material was considered to be moderately irritating.
Executive summary:

The skin irritation potential of the test material was investigated in a patch test.

The test was performed using 8 New Zealand White rabbits, 0.5 mL of undiluted test material was applied to the clipped dorsum of the rabbits for 4 hours under a semi-occlusive patch. The irritation reactions were assessed immediately after removal of the patch and after 24, 48 and 72 hours.

The test material produced distinct erythema and oedema at 4 hours which intensified slightly to show variable but generally slight/moderate effects 72 hours after treatment. Two animals developed extensive areas of brown tissue.

Under the conditions of this study the test material was considered to be moderately irritating.

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
25 February 1985 to 03 April 1985
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
no guideline followed
Principles of method if other than guideline:
The rabbit eye irritation test according to test method No. OCTM05.01 dated 4.2.82 but with the following modifications:
- Treatment groups of the test material were diluted in Tween 80 (normally 10 and 50% dilutions) were omitted in this test.
- Only one animal was treated.
The study was conducted in accordance with S.O.P. 013
GLP compliance:
not specified
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Commercial Supplier
- Age at study initiation: 9 weeks old
- Weight at study initiation: 2.3 kg
- Housing: animals were housed individually in cages with wire-mesh floors
- Diet: certified pelleted commercial rabbit diet ad libitum
- Water: ad libitum
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 100 µL
- Concentration: undiluted

Duration of treatment / exposure:
The test material was applied to the conjunctival sac formed by gently pulling the lower eyelid away from the eyeball. After treatment the eye was closed and the eyelids released.
Observation period (in vivo):
8 days
Number of animals or in vitro replicates:
1
Details on study design:
- Eyes were examined before application of the test material and at daily intervals afterwards with a slit lamp and corneal swelling was measured.
- Animals were observed carefully during and immediately after treatment for evidence of pain or discomfort. If excessive or persistent the substance was immediately washed out with copious quantities of physiological saline
- The eye was examined 15 minutes after treatment and thereafter at 2, 3, 4, 5 and 8 days and graded for corneal, conjunctival and iridial damage.


SCORING SYSTEM:

Cornea:
Normal = 0
Any change from normal, including slight dulling of corneal lustre = 0.5
Scattered or diffuse areas of opacity, or loss of corneal epithelium. Iris clearly visible = 1
Easily discernible, greyish, translucent areas, details of iris slightly obscured = 2
Grey-white areas, no details of iris visible, size of pupil barely discernible = 3
Complete corneal opacity, iris not discernible = 4

Iris
Normal = 0
Any divergence from normal, congestion of iridial vessels, deepening of folds = +
Severe congestion of iridial vessels or deepening of folds. Loss of iridial reflex to light = ++

Area of cornea or iris affected
Less than 1/4 of area affected = 1
Between 1/4 and 1/2 of area affected = 2
Between 1/2 and 3/4 of area affected = 3
Between 3/4 and whole area affected = 4

Conjunctivae
Redness (palpebral and bulbar conjunctivae and nictitating membrane)
Vessels normal = 0
Some vessels definitely injected = 1
Diffuse, crimson red, individual vessels not easily discernible = 2
Diffuse, beefy red = 3

Chemosis
No swelling = 0
Any swelling above normal = 1
Obvious swelling with partial eversion of eyelids = 2
Swelling with lids about half closed = 3
Swelling with lids more than half closed = 4

Discharge
No discharge = 0
Any discharge different from normal = 1
Discharge with moistening of lids and hairs just adjacent to the lids = 2
Discharge with moistening of lids and considerable area round eye = 3

Pannus

Healing
The insertion of 'N' as a score of corneal opacity denotes that the whole eye had returned to normal on the day indicated.

Corneal Swelling
Corneal thickness is measured with a slit-lamp 24 hours after treatment and thereafter at daily intervals until the thickness has returned to its pretreatment value. Slit-lamp readings are recorded in the Eye Treatment Book and the percentage corneal swelling is calculated from:
corneal swelling = (corneal thickness on day of measurement /corneal thickness before treatment) -1 x 100

TOOL USED TO ASSESS SCORE: hand-slit lamp
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.7
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.7
Max. score:
3
Reversibility:
fully reversible within: 8 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.3
Max. score:
4
Reversibility:
fully reversible within: 5 days
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.3
Max. score:
4
Reversibility:
fully reversible within: 3 days
Irritant / corrosive response data:
One rabbit only was treated. The eye subsequently showed extensive loss of corneal epithelium with moderate corneal swelling and iritis. Conjunctivitis was moderate. The eye had healed normally by day 8. The test material is a moderate eye irritant at 100 μL dosage.

Table 1: Summary of Observations for the Test Material Treated Animal

Observations

Discomfort

Conjunctival Reactions

Iritis

Pannus

Maximum Corneal Opacity (Max. 4)

Maximum Corneal Swelling

Yes

Yes

Yes

No

1

Moderate

(46 – 80 %)

Table 2: Daily Observations

Animal

Region of Eye

Day

15 min

2

3

4

5

8

1

Cornea

Opacity

-

1

0.5

0.5

0.5

N

Area

-

1

1

1

1

-

Swelling (%)

-

71

15

0

-

-

Conjunctivae

Redness

2

2

2

1

1

0

Chemosis

2

2

1

1

0

0

Discharge

-

1

0

0

0

0

Iris

-

+4

0

0

0

0

Interpretation of results:
study cannot be used for classification
Conclusions:
Under the conditions of this study the test material was considered to be a moderate eye irritant.
Executive summary:

The eye irritation potential of the test material was examined using a New Zealand White rabbit.

The test material was applied to the conjunctival sac formed by gently pulling the lower eyelid away from the eyeball. After treatment the eye was closed and the eyelids released.

One rabbit only was treated. The eye subsequently showed extensive loss of corneal epithelium with moderate corneal swelling and iritis. Conjunctivitis was moderate. The eye had healed normally by day 8.

Under the conditions of this study the test material was considered to be a moderate eye irritant.

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 February 1985 to 03 April 1985
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
no guideline followed
Principles of method if other than guideline:
Modified Federal Hazardous Substances Labelling Act Method.
Test substances are applied to one eye of each of two rabbits by gently pulling the lower lid away from the eye ball and placing 10 μL in the sac so formed.
Eyes are examined 24 hours after treatment and thereafter at daily intervals and graded for corneal, conjunctival and iridial damage. Eyes are examined before application of materials and at daily intervals afterwards with a slit lamp and corneal swelling is measured.
- Modification to standard test method: only 2 animals per test group
The study was conducted in accordance with S.O.P. 013.
GLP compliance:
not specified
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Commercial Supplier
- Age at study initiation: 9 weeks old
- Weight at study initiation: 1.7 - 2.0 kg
- Housing: The animals are housed individually in cages with wire-mesh floors
- Diet: certified pelleted commercial rabbit diet ad libitum
- Water: ad libitum
Vehicle:
other: Tween 80
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 10 µL
- Concentration: Group A: 50% concentration un Tween 80, Group B: undiluted

Duration of treatment / exposure:
The test material was applied to the conjunctival sac formed by gently pulling the lower eyelid away from the eyeball. After treatment the eye was closed and the eyelids released.
Observation period (in vivo):
21 days
Number of animals or in vitro replicates:
2 males per dose
Details on study design:
- The eye was examined 24 hours after treatment and thereafter at daily intervals and graded for corneal, conjunctival and iridial damage.
- Eyes were examined before application of the test material and at daily intervals afterwards with a slit lamp and corneal swelling was measured.

SCORING SYSTEM:

Cornea:
Normal = 0
Any change from normal, including slight dulling of corneal lustre = 0.5
Scattered or diffuse areas of opacity, or loss of corneal epithelium. Iris clearly visible = 1
Easily discernible, greyish, translucent areas, details of iris slightly obscured = 2
Grey-white areas, no details of iris visible, size of pupil barely discernible = 3
Complete corneal opacity, iris not discernible = 4

Iris
Normal = 0
Any divergence from normal, congestion of iridial vessels, deepening of folds = +
Severe congestion of iridial vessels or deepening of folds. Loss of iridial reflex to light = ++

Area of cornea or iris affected
Less than 1/4 of area affected = 1
Between 1/4 and 1/2 of area affected = 2
Between 1/2 and 3/4 of area affected = 3
Between 3/4 and whole area affected = 4

Conjunctivae
Redness (palpebral and bulbar conjunctivae and nictitating membrane)
Vessels normal = 0
Some vessels definitely injected = 1
Diffuse, crimson red, individual vessels not easily discernible = 2
Diffuse, beefy red = 3

Chemosis
No swelling = 0
Any swelling above normal = 1
Obvious swelling with partial eversion of eyelids = 2
Swelling with lids about half closed = 3
Swelling with lids more than half closed = 4

Discharge
No discharge = 0
Any discharge different from normal = 1
Discharge with moistening of lids and hairs just adjacent to the lids = 2
Discharge with moistening of lids and considerable area round eye = 3

Pannus
- Severity
0 = No vessels growing into the cornea
1 = Vessels just onto the cornea
2 = Vessels quite clearly onto the cornea, but less than half-way across the iris.
3 = Vessels more than half-way across the iris
4 = Vessels extending across to the pupil

- Extent:
1 = Four or less vessels
2 = A band of vessels affecting up to 1/4 of the corneal circumference
3 = A band of vessels affecting more than 1/4 but less than 1/2 the corneal circumference
4 = A band of vessels affecting more than 1/2 the corneal circumference

- Blood Content of Vessels:
1 = All the vessels empty
2 = More than half but not all, the vessels empty
3 = More than half, but not all, the vessels full
4 = All the vessels full

Healing
The insertion of 'N' as a score of corneal opacity denotes that the whole eye had returned to normal on the day indicated.

Corneal Swelling
Corneal thickness is measured with a slit-lamp 24 hours after treatment and thereafter at daily intervals until the thickness has returned to its pretreatment value. Slit-lamp readings are recorded in the Eye Treatment Book and the percentage corneal swelling is calculated from:
corneal swelling = (corneal thickness on day of measurement /corneal thickness before treatment) -1 x 100

TOOL USED TO ASSESS SCORE: hand-slit lamp
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
50 % in Tween 80
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not fully reversible within: 5 days
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
50 % in Tween 80
Time point:
24/48/72 h
Score:
4
Max. score:
4
Reversibility:
not fully reversible within: 5 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #1
Remarks:
50 % in Tween 80
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
not fully reversible within: 5 days
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
50 % inb Tween 80
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 5 days
Irritation parameter:
cornea opacity score
Basis:
animal #2
Remarks:
50 % in Tween 80
Time point:
24/48/72 h
Score:
1.3
Max. score:
4
Reversibility:
not fully reversible within: 22 days
Irritation parameter:
iris score
Basis:
animal #2
Remarks:
50 % in Tween 80
Time point:
24/48/72 h
Score:
4
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #2
Remarks:
50 % in Tween 80
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 15 days
Irritation parameter:
chemosis score
Basis:
animal #2
Remarks:
50 % in Tween 80
Time point:
24/48/72 h
Score:
1.7
Max. score:
4
Reversibility:
fully reversible within: 10 days
Irritant / corrosive response data:
Two rabbits were treated with 10 µL of 50 % test material in Tween 80. Both eyes subsequently showed an almost total loss of corneal epithelium with diffuse corneal opacity and severe corneal swelling. Both eyes showed iritis. Conjunctivitis was moderate with copious discharge. Both eyes developed pannus. One animal was killed at day 5 owing to the severity of these reactions. The eye of the remaining animal developed small areas of persistent corneal opacity.
Two rabbits were treated with 10 µL of undiluted test material. Both eyes showed only a slight transient conjunctivitis which had healed by day 2.

Table 1: Daily Observations in Group A (50 % in Tween 80)

Animal

Region of Eye

Day

15 min

2

3

4

5

8

10

12

15

19

22

1

Cornea

Opacity

-

1

1

1

3

 

 

 

 

 

 

Area

-

4

4

3

1

 

 

 

 

 

 

Swelling (%)

-

66

66

NM

NM

 

 

 

 

 

 

Conjunctivae

Redness

2

2

2

2

2

 

 

 

 

 

 

Chemosis

1

2

2

2

1

 

 

 

 

 

 

Discharge

-

20

20

20

10

 

 

 

 

 

 

Iris

-

+4

+4

+4

+4

 

 

 

 

 

 

2

Cornea

Opacity

-

1

1

2

2

2

2

2

2

2

2

Area

-

4

4

3

3

1

1

1

1

1

1

Swelling (%)

-

62

66

NM

NM

23

0

 

 

 

 

Conjunctivae

Redness

2

2

2

2

2

1

1

1

0

0

0

Chemosis

1

2

2

1

1

1

0

0

0

0

0

Discharge

-

3

3

10

10

10

0

0

0

0

0

Iris

-

+4

+4

+4

+4

0

0

0

0

0

0

Table 2: Daily Observations in Group B (undiluted)

Animal

Region of Eye

Day

15 min

2

3

4

5

3

Cornea

Opacity

-

N

 

 

 

Area

-

 

 

 

 

Swelling (%)

-

0

 

 

 

Conjunctivae

Redness

1

0

 

 

 

Chemosis

1

0

 

 

 

Discharge

-

0

 

 

 

Iris

-

0

 

 

 

4

Cornea

Opacity

-

N

 

 

 

Area

-

 

 

 

 

Swelling (%)

-

0

 

 

 

Conjunctivae

Redness

2

0

 

 

 

Chemosis

1

0

 

 

 

Discharge

-

0

 

 

 

Iris

-

0

 

 

 

Interpretation of results:
study cannot be used for classification
Conclusions:
Under the conditions of this study 10 μL of 50 % test material was considered to produce moderate/severe irritation, 10 μL of undiluted test material was considered to produce slight irritation.
Further testing at 100 μL dosage of the undiluted material is therefore required.
Executive summary:

The eye irritation potential of the test material was examined using New Zealand White rabbits.

The test material was applied to one eye of each of two rabbits by gently pulling the lower lid away from the eye ball and placing 10 μL in the sac so formed. Eyes were examined 24 hours after treatment and thereafter at daily intervals and graded for corneal, conjunctival and iridial damage. Eyes were examined before application of materials and at daily intervals afterwards with a slit lamp and corneal swelling was measured.

Two rabbits were treated with 10 µL of 50 % test material in Tween 80. Both eyes subsequently showed an almost total loss of corneal epithelium with diffuse corneal opacity and severe corneal swelling. Both eyes showed iritis. Conjunctivitis was moderate with copious discharge. Both eyes developed pannus. One animal was killed at day 5 owing to the severity of these reactions. The eye of the remaining animal developed small areas of persistent corneal opacity.

Two rabbits were treated with 10 µL of undiluted test material. Both eyes showed only a slight transient conjunctivitis which had healed by day 2.

Under the conditions of this study 10 μL of 50 % test material was considered to produce moderate/severe irritation, 10 μL of undiluted test material was considered to produce slight irritation.

Further testing at 100 μL dosage of the undiluted material is therefore required.

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Irritation, York (1985)

The skin irritation potential of the test material was investigated in a patch test. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).

The test was performed using 8 New Zealand White rabbits, 0.5 mL of undiluted test material was applied to the clipped dorsum of the rabbits for 4 hours under a semi-occlusive patch. The irritation reactions were assessed immediately after removal of the patch and after 24, 48 and 72 hours.

The test material produced distinct erythema and oedema at 4 hours which intensified slightly to show variable but generally slight/moderate effects 72 hours after treatment. Two animals developed extensive areas of brown tissue.

Under the conditions of this study the test material was considered to be moderately irritating.

Eye Irritation RE 85.05 (1985)

The eye irritation potential of the test material was examined using New Zealand White rabbits. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).

The test material was applied to one eye of each of two rabbits by gently pulling the lower lid away from the eye ball and placing 10 μL in the sac so formed. Eyes were examined 24 hours after treatment and thereafter at daily intervals and graded for corneal, conjunctival and iridial damage. Eyes were examined before application of materials and at daily intervals afterwards with a slit lamp and corneal swelling was measured.

Two rabbits were treated with 10 µL of 50 % test material in Tween 80. Both eyes subsequently showed an almost total loss of corneal epithelium with diffuse corneal opacity and severe corneal swelling. Both eyes showed iritis. Conjunctivitis was moderate with copious discharge. Both eyes developed pannus. One animal was killed at day 5 owing to the severity of these reactions. The eye of the remaining animal developed small areas of persistent corneal opacity.

Two rabbits were treated with 10 µL of undiluted test material. Both eyes showed only a slight transient conjunctivitis which had healed by day 2.

Under the conditions of this study 10 μL of 50 % test material was considered to produce moderate/severe irritation, 10 μL of undiluted test material was considered to produce slight irritation.

Further testing at 100 μL dosage of the undiluted material is therefore required.

Eye Irritation RE 85.09 (1985)

The eye irritation potential of the test material was examines using a New Zealand White rabbit. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).

During the study 100 µL of undiluted test material was applied to the conjunctival sac formed by gently pulling the lower eyelid away from the eyeball. After treatment the eye was closed and the eyelids released.

One rabbit only was treated. The eye subsequently showed extensive loss of corneal epithelium with moderate corneal swelling and iritis. Conjunctivitis was moderate. The eye had healed normally by day 8.

Under the conditions of this study the test material was considered to be a moderate eye irritant.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No. 1272/2008, the substance does not require classification with respect to skin corrosion/ irritation. The substance is, however classified as an eye irritant (category 2) on a precautionary basis and is assigned the hazard statement H319: Causes serious eye irritation.