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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From March 22, 2004 to March 22, 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
PARAD substance 139
Specific details on test material used for the study:
- Name of the test substance (as cited in the report): PARAD Substance 139.
- Batch: 31104251.
- Molecular formula: C6H10O3.xH3O4P.
- Molecular weight: ~250.
- CAS number: 52628-03-2.
- Description: Slightly yellowish viscous liquid.
- Purity: Not available (mixture).
- Storage: At room temperature in the dark.
- Stability under storage conditions: Stable
- Expiry date: December 31, 2004.

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
- Age and body weight: The animal used within the study was at least 6 weeks old and its body weight was between 1.0 and 3.5 kg.
- Identification: Earmark.
- Conditions: Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per h, a temperature of 21.0±3.0°C (actual range: 16.6 - 21.6°C), a relative humidity of 30 - 70% (actual range: 33 - 62%) and 12 h artificial fluorescent light and 12 h darkness per day.
- Accommodation: Individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56x44x37.5 cm). Acclimatisation period was at least 5 d before start of treatment under laboratory conditions.
- Diet: Standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Lage, Germany) approximately 100 g per d. In addition, hay (BMI, Helmond, the Netherlands) was provided at least three times a week.
- Water: ad libitum.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: The other eye remained untreated and served as the reference control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 0.1 mL
Duration of treatment / exposure:
1 h
Observation period (in vivo):
1 h
Number of animals or in vitro replicates:
1
Details on study design:
- A health inspection was performed prior to commencement of treatment, to ensure that the animal was in a good state of health. Special attention was paid to the eyes, which were free from any abnormality.

- On test Day 1, 0.1 mL of the test substance as the maximum required amount, was instilled in the conjunctival sac of an eye of one animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control. Based on the severity of the ocular lesions observed during the study, the animal was sacrificed for ethical reasons immediately after the 1 h observation and the two further rabbits assigned to the study were not treated.

- Observations:
Mortality/Viability: Twice daily.
Toxicity: At least once daily.
Body Weight: Day of treatment (prior to instillation).
Irritation: The eyes of the animal were examined approximately 1 h after instillation of the test substance. The irritation scores and a description of all other (local) effects were recorded.

- The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

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

Area of cornea involved:
No ulceration or opacity ............................................................................................................... 0
One quarter or less but not zero .................................................................................................. 1
Greater than one quarter, but less than half ................................................................................. 2
Greater than half, but less than three quarters ............................................................................. 3
Greater than three quarters, up to whole area .............................................................................. 4

IRIS:
Normal ........................................................................................................................................ 0
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) ....................................................................................................... 1
No reaction to light, hemorrhage, gross destruction (any or all of these) ...................................... 2

CONJUNCTIVAL IRRITATION:
Redness (refers to palpebrae and sclera, excluding cornea and iris):
Blood vessels normal .................................................................................................................. 0
Some blood vessels definitely hyperaemic (injected) ................................................................... 1
Diffuse, crimson color, individual vessels not easily discernible .................................................... 2
Diffuse beefy red .......................................................................................................................... 3

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

Discharge:
No discharge (may include small amounts observed in inner canthus of normal animals) ........... 0
Any amount different from normal and/or lacrimation ................................................................... 1
Discharge with moistening of the lids and hairs just adjacent to lids ............................................. 2
Discharge with moistening of the lids and hairs (considerable area around the eye) ................... 3
Where standard lighting was considered inadequate for observing minor effects, eye examinations were performed using an ophthalmic examination lamp.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: 1 hr
Score:
ca. 3
Max. score:
3
Reversibility:
not reversible
Remarks on result:
other: Based on the degree of the ocular injury, it was concluded that ocular corrosion had occurred into the treated eye of the animal. Based on animal welfare reasons, no other animals were tested.
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: 1 hr
Score:
ca. 1
Max. score:
1
Reversibility:
not reversible
Remarks on result:
other: Based on the degree of the ocular injury, it was concluded that ocular corrosion had occurred into the treated eye of the animal. Based on animal welfare reasons, no other animals were tested.
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: 1 hr
Score:
ca. 3
Max. score:
3
Reversibility:
not reversible
Remarks on result:
other: Based on the degree of the ocular injury, it was concluded that ocular corrosion had occurred into the treated eye of the animal. Based on animal welfare reasons, no other animals were tested.
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 1 hr
Score:
ca. 2
Max. score:
2
Reversibility:
not reversible
Remarks on result:
other: Based on the degree of the ocular injury, it was concluded that ocular corrosion had occurred into the treated eye of the animal. Based on animal welfare reasons, no other animals were tested.
Irritant / corrosive response data:
Irritation and corrosion:
Instillation of 0.1 mL of test substance into an eye of one rabbit resulted in effects on the cornea, iris and conjunctivae. The corneal injury consisted of opacity (maximum grade 3). Iridial irritation grade 1 was observed. The irritation of the conjunctivae consisted of redness, chemosis and discharge. Gray/white discoloration (signs of necrosis) was noted on the eyelids and nictitating membrane, 1 h after instillation of the test substance. Based on the severity of the ocular lesions observed during the study, the animal was sacrificed for ethical reasons immediately after the 1 h observation.

Colouration / remnants:
No staining of (peri) ocular tissues by the test substance was observed and no test substance remnants were seen.
Other effects:
Toxicity / mortality:
No symptoms of systemic toxicity were observed in the animal during the test period and no mortality occurred.

Any other information on results incl. tables

Table 1: Individual eye irritation scores

 

Time after

dosing

Cornea

Iris

Conjunctivae

Opacity

Area

Fluor

area (%)

Redness

Chemosis

Discharge

Comments

1 h

3

2

-

1

3

2

2

a, b

 

Fluor area (%): green staining (percentage of total corneal area) after fluorescein treatment.

Comments:

(a) Signs of necrosis were noted on the eyelids and nictitating membrane.

(b) Based on the severity of the ocular lesions observed during the study, the animal was sacrificed for ethical reasons immediately after the 1 h observation.

Applicant's summary and conclusion

Interpretation of results:
other: Category 1
Remarks:
as per CLP criteria
Conclusions:
Under the study conditions, the test substance was found to cause corrosive effects on the eyes.
Executive summary:

A study was conducted to assess the eye irritancy potential of the test substance in rabbit according to OECD Guideline 405, EU Method B.5, US EPA OPPTS 870.2400 and JMAFF Guideline, 2000, in compliance with GLP. A single dose of 0.1 mL test substance was instilled into the sac of one eye. The lids were then gently held together for approximately one second to prevent loss of the test substance. Gray/white discoloration (signs of necrosis) was noted on the eyelids and nictitating membrane, 1 h after instillation of the test substance. Based on the severity of the ocular lesions observed during the study, the animal was sacrificed for ethical reasons immediately after the 1 h observation. The remaining two rabbits assigned to the study were not treated. Based on the degree of the corneal injury, it was concluded that ocular corrosion had occurred by instillation of test substance into the rabbit eye in the animal. Hence, the substance was found to cause corrosive effects on the eyes (van Huygevoort AHBM, 2004).