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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:
November 2013 - February 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD guidelines and according to GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
(2012)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Version / remarks:
(1998)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Specific details on test material used for the study:
- Name of test material (as cited in study report): EXP1200078
- Physical state: dark brown viscous liquid
- Storage condition of test material: at room temperature in the dark

- Analytical purity: 100%
- Lot/batch No.: E00275-350
- Expiration date of the lot/batch:End-2015

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
- Source: Covance Research Products, Inc., Greenfield, IN, USA
- Age at study initiation: Animals used within the study were approx 9 months old.
- Weight at study initiation: Body weights were 3321-3459 g.
- Housing: Individually housed in stainless steel cages.
- Diet: The basal diet for rabbits (PMI Nutrition International, LLC, Certified Rabbit HF LabDiet® 5325)was provided at approx 150 g/day.
- Water: Free access to tap water.
- Acclimation period: At least 5days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.5-18.7
- Humidity (%): 45.4-53.1
- Air changes (per hr): approx 10
- Photoperiod (hrs dark / hrs light): 12/12

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 applied: 0.1 mL (liquid)
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 dosing, and on days 4, 7, 10 if irritation persisted.
Number of animals or in vitro replicates:
3 males
Details on study design:
TREATMENT
Approximately 60 minutes prior to test substance application, buprenorphine (0.01 mg/kg) was administered by subcutaneous injection (SC) to provide a therapeutic level of systemic analgesia.
Approximately 5 minutes prior to test substance application, one drop of a topical ocular anesthetic (0.5% proparacaine hydrochloride) was applied to each eye. The test substance (0.1 mL) was placed directly into the cupped lower conjunctival sac of each rabbit’s right (test) eye. The eyelid was held closed for approximately 1 second after instillation and then released. The left eye of each animal was manipulated in an identical manner to simulate the dosing of the right eye. Following test substance administration, no additional topical anesthetics were applied in order to avoid interference with the study.
Initially, a single animal was dosed. Based on the results 2 additional rabbits were exposed.
Approximately 8 hours after test substance administration, buprenorphine (0.01 mg/kg) SC and meloxicam (0.5 mg/kg) SC were administered to provide a continued therapeutic level of systemic analgesia. Following the initial 8-hour post-dosing treatment, buprenorphine (0.01 mg/kg) SC was administered every twelve (12) hours, in conjunction with meloxicam (0.5 mg/kg) SC every 24 hours, until positive ocular lesions/irritation resolved and no clinical signs of pain and distress were detected.

REMOVAL OF TEST SUBSTANCE
-Washing: No

OBSERVATIONS (beschrijf beperkt)
- Mortality/Viability: Twice daily.
- Toxicity: At least once daily.
- Body Weight: Obtained on study day 0 (initiation) and at each rabbit’s termination from the study (study day 7 or 10).
- Necropsy: the rabbits were euthanized by intravenous injection of sodium pentobarbital and discarded.
- Irritation: prior to initiation of dosing. The pre-initiation examination included the use of sodium fluorescein and a direct ophthalmoscope for detection of corneal abnormalities. Only rabbits with no pre existing ocular abnormalities were assigned to study. Both eyes of all rabbits were examined macroscopically for ocular irritation in accordance with the method of Draize at approximately 1, 24, 48 and 72 hours, and days 4, 7, 10 after instillation of the test substance when irritation persisted. The irritation scores and a description of all other (local) effects were recorded.The irritation was assessed according to OECD 405.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.33
Max. score:
3
Reversibility:
fully reversible within: 10
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 2 days
Irritant / corrosive response data:
Conjunctival irritation was noted for all 3 rabbits. The only grade 2 finding was observed for one male at 24 hours post-dosing. All other conjunctival irritation was graded as 1 and subsided by study day 10.
The left (control) eyes were free of evidence of ocular irritation and other findings for the duration of the study.
Other effects:
One animal had light brown staining of hair surrounding the eye. The other 2 animals had dried light brown material around eye.

There were no deaths during the study, nor test substane related clinical observations.
There were no remarkable body weight changes noted during the study. One rabbit had a very slight body weight loss (0.8%) from study days 0 to 7. However, body weight losses are commonly associated with animals that are on a restricted diet and can be enhanced by the use of systemic analgesics.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an eye irritation study with rabbits, performed according to OECD test guidelines, slight irritation was observed, which disappeared by day 10.
Executive summary:

In an eye irritation study with rabbits, performed according to OECD and OPPTS test guidelines, the eyes of 3 rabbits were exposed to 0.1 mL of the test substance. Conjunctival irritation was noted for all 3 rabbits. The only grade 2 finding was observed for one male at 24 hours post-dosing. All other conjunctival irritation was graded as 1 and subsided by day 10. Therefore, the substance is considered to be not an irritant for the eyes.