N,N''-(methylenedi-4,1-phenylene)bis[N'-octyl]urea

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October 17, 2002 - November 07, 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

451-060-3

EC Name:

-

Cas Number:

122886-55-9

Molecular formula:

Hill formula: C31H48N4O2 CAS formula: C31H48N4O2

IUPAC Name:

N,N’’-(methylenedi-4,1-phenylene)bis(N’-octylurea)

Constituent 2

Reference substance name:

N-{4-[(4-{[(octadecylamino)carbonyl]amino}phenyl)methyl]phenyl}-N’-octylurea

Cas Number:

117328-86-6

Molecular formula:

Hill formula: C41H68N4O2 CAS formula: C41H68N4O2

IUPAC Name:

N-{4-[(4-{[(octadecylamino)carbonyl]amino}phenyl)methyl]phenyl}-N’-octylurea

Constituent 3

Reference substance name:

3,3'-dioctadecyl-1,1'-methylenebis(4,1-phenylene)diurea

EC Number:

406-690-3

EC Name:

3,3'-dioctadecyl-1,1'-methylenebis(4,1-phenylene)diurea

Cas Number:

43136-14-7

Molecular formula:

Hill formula: C51H88N4O2 CAS formula: C51H88N4O2

IUPAC Name:

N,N’’-(methylenedi-4,1-phenylene)bis(N’-octadecylurea)

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material (as cited in study report): KY-EU
- Appearance: white powder
- Storage condition of test material: in darkness at room temperature

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: CEGAV, Saint Mars d'Egrenne, France
- Age at study initiation: 2 to 4 months old
- Weight at study initiation: 2.9 ± 0.2 kg
- Housing: Individually housed in polystyrene cages (48.2 cm x 58 cm x 36.5 cm).
- Diet (e.g. ad libitum): Free access to 110 pelleted diet (UAR, Villemoisson, Epinay-sur-Orge, France).
- Water (e.g. ad libitum): Free access to water filtered by a FG Millipore membrane (0.22 micron).
- Acclimatization period: at least 5 days before start of treatment.

Food was analysed regularly by the supplier for composition and contaminant levels. Bacteriological and chemical analyses of water were performed regularly by external laboratories. These analyses include the detection of possible contaminants (pesticides, heavy metals and nitrosamines).
No contaminants were known to have been present in the diet or drinking water at levels which may be expected to have interfered with or prejudiced the outcome of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3
- Humidity (%): 30 - 70
The relative humidity recorded in the animal room was sometimes outside of the target ranges specified in the study plan. This minor deviation was not considered to have compromised the validity or integrity of the study.
- Air changes (per hr): approximately 12
- Photoperiod (hrs dark / hrs light): 12/12

IN LIFE DATES: From: October 17, 2002 To: November 07, 2002

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: One eye of each animal remained untreated and served as the reference control.

Amount / concentration applied:

TEST MATERIAL- Amount(s) applied (volume or weight with unit): 100 mg

Duration of treatment / exposure:

Single instillation on Day 1.

Observation period (in vivo):

The eyes of each animal were examined approximately 1, 24, 48 and 72 hours after instillation of the test substance. Following the guidelines, since there was persistent ocular reactions at 72 hours, the observation period was extended. In the absence of complete reversibility of ocular reactions on day 22, the study was ended.

Number of animals or in vitro replicates:

3 males

Details on study design:

TREATMENT
The day before treatment, the eyes of each animal were examined in order to check the absence of any signs of ocular irritation, ocular defects or pre-existing corneal injury.
As no irritant effects were anticipated, the study was directly performed with three animals.
A single dose of 100 mg of the test item in its original form was introduced into the conjunctival sac of the left eye after gently pulling the lower lid away from the eyeball.
The lower and upper eyelids were held together for about one second to avoid any loss of test item.

REMOVAL OF TEST SUBSTANCE
-Washing: No

OBSERVATIONS
- Mortality/Viability: No data
- Toxicity: No data
- Body Weight: No data
- Necropsy: No data
- Irritation: The eyes were examined approximately 1 hour, 24, 48 and 72 hours after administration of the test item. Following the guidelines, since there was persistent ocular reactions at 72 hours, the observation period was extended. In the absence of complete reversibility of ocular reactions on day 22, the study was ended.

Ocular reactions were scored according to the following numerical scale:

CORNEAL LESIONS
Opacity: degree of density (area most dense taken for reading)
0: No ulceration or opacity
1: Scattered or diffuse areas of opacity (other than slight dulling or normal lustre), details of iris clearly visible
2: Easily discernible translucent area, details of iris slightly obscured
3: Nacreous area, no details of iris visible, size of pupil barely discernible
4: Opaque cornea, iris not discernible through the opacity

Area of opacity:
1: One quarter (or less) but not zero
2: Greater than one quarter, but less than half
3: Greater than one half, but less than three quarters
4: Greater than three quarters, up to whole area

IRIS
0: Normal
1: Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection, any of these or combination thereof, iris still reacting to light (sluggish reaction is positive)
2: No reaction to light, hemorrhage, gross destruction (any or all of these)

CONJUNCTIVAL IRRITATION
Redness (refers to palpebral and bulbar conjunctivae, cornea and iris):
0: Blood vessels normal
1: A number of blood vessels definitely hyperaemic (injected)
2: Diffuse, crimson color, individual vessels not easily discernible
3: Diffuse beefy red

Chemosis (lids and/or nictitating membranes):
0: No swelling
1: Any swelling above normal (includes nictitating membranes)
2: Obvious swelling with partial eversion of lids
3: Swelling with lids about half closed
4: Swelling with lids more than half closed

Discharge:
0: Absence of discharge
1: Slight discharge (does not include small amounts normally found in inner canthus)
2: Discharge with moistening of the lids and hairs just adjacent to lids
3: Discharge with moistening of the lids and hairs on wide area around eye

Any other lesions observed were recorded.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 5 days

Results 2

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 4 days

Results 3

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 3 days

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.7

Max. score:

2

Reversibility:

fully reversible within: 4 days

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 7

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.3

Max. score:

3

Reversibility:

fully reversible within: 6 days

Remarks on result:

other: Whitish purulent discharge was observed on day 3.

Results 8

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #2

Time point:

24/48/72 h

Score:

2.3

Max. score:

3

Reversibility:

not fully reversible within: 21 days

Remarks on result:

other: Whitish purulent discharge was observed from day 3-8.

Results 9

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.3

Max. score:

3

Reversibility:

fully reversible within: 5 days

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.7

Max. score:

4

Reversibility:

fully reversible within: 6 days

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2.3

Max. score:

4

Reversibility:

not fully reversible within: 21 days

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 5 days

Irritant / corrosive response data:

Individual and mean eye irritation scores are specified in the attached tables.

Instillation of the test substance into one eye of each of three rabbits resulted in ocular reactions; irritation of the conjunctivae, which was seen as redness and chemosis; a slight iritis was noted in one animal on days 2 and 3 only; a slight corneal opacity was recorded in all animals on day 2, it persisted up to day 2 (one animal) or 4 (one animal). In animal #2, the effects on the conjunctivae score (redness) and chemosis score were not fully reversed within the observation period of 21 days. In the other animals, the observed effects were fully reversed within 21 days.

Other effects:

Residual of the test item was observed in all animals 1 and 24 hours after instillation.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

An acute eye irritation/corrosion study with KY-EU in the rabbit (3 males) was conducted according to OECD 405 and GLP guidelines. Based on the results, KY-EU should be classified as serious eye damage cat.1 and labelled with H318: Causes serious eye damage.

Executive summary:

An acute eye irritation/corrosion study with KY-EU in the rabbit (3 males) was conducted according to OECD 405 and GLP guidelines.

Instillation of the test substance into one eye of each of three rabbits resulted in ocular reactions; irritation of the conjunctivae, which was seen as redness and chemosis; a slight iritis was noted in one animal on days 2 and 3 only; a slight corneal opacity was recorded in all animals on day 2, it persisted up to day 2 (one animal) or 4 (one animal). In animal #2, the effects on the conjunctivae score (redness) and chemosis score were not fully reversed within the observation period of 21 days. In the other animals, the observed effects were fully reversed within 21 days.

Based on the results, KY-EU should be classified as serious eye damage cat.1 and labelled with H318: Causes serious eye damage according to Regulation (EC) No 1272/2008.