Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
3 November 1997 - 6 November 1997
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to OECD Guideline 405 and EU Method B.5 (Acute Eye Irritation/Corrosion) and GLP practices.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report Date:
1998

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
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid - liquid: suspension
Details on test material:
- Name of test material (as cited in study report): Epyrrol
- Substance type: pure active substance
- Physical state: Powder, off white in color
- Composition of test material, percentage of components: Main component= 75%, other= 18%, other= 7%
- Lot/batch No.: GL-1291 K1
- Expiration date of the lot/batch: 24 September 1998
- Stability under test conditions: Not indicated
- Storage condition of test material: Stable at room temperature in the dark

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Broekman Institute, Someren, The Neitherlands
- Age at study initiation: At least six weeks old
- Weight at study initiation: Less then 3.5 kg
- Housing: Individually in labelled cages with perforated floors
- Diet (e.g. ad libitum): Standard laboratory rabbit diet (LKK-20, pellet diameter 4mm., Hope Farms, Woerden, The Neitherlands), approximately 100 grams/day. Additionally, hay was provided once a week.
- Water (e.g. ad libitum): Free access to tap-water diluted with decalcified water
- Acclimation period: At least 5 days before the start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 deg C
- Humidity (%): 50%
- Air changes (per hr): 15 per hour
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: Not provided in study report To: Not provided in study report

Test system

Vehicle:
not specified
Controls:
other: Treatment was only applied to one eye, the other eye served as the control for each animal.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 29.9 +/- 0.3 mL (0.1 mL volume)
Duration of treatment / exposure:
Single application
Observation period (in vivo):
Up to 72 hours
Number of animals or in vitro replicates:
3 male rabbits
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): A solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled in both eyes of each animal.
- Time after start of exposure: 24 hours after treatment

SCORING SYSTEM: Evaluation of eye reactions were based on a numerical scoring system used to score eye lesions, evaluating the corneal irritation (scale of 0-4), iris (scale of 0-2), conjunctival irritation (scale of 0-3), chemosis (scale of 0-4), and discharge (scale of 0-3), with 0 being the low end of the scoring system and/or no effects.

TOOL USED TO ASSESS SCORE: Fluorescein, although in case standard lighting was considered inadequate for observing minor effects, eye examinations were performed using an ophthalmic examination lamp.

Results and discussion

In vivo

Irritant / corrosive response data:
Installation of approximately 30 mg of Epyrrol (a volume of ~ 0.1 mL) into one eye of each of three rabbits resulted in effects on the cornea, iris, and conjunctivae.

The Corneal injury consisted of opacity (maximum grades 2-3) and epithelial damage (maximum 75% of the corneal area).

Iridic irritation grade 1 was observed in all animals.

In all animals, approximately 25% of the cornea and iris could not be scored, due to oedema of the eyelids, 24 and/or 48 hours after instillation.

In one animal, approximately 35% of the iris could not be scored after 48 and 72 hours, due to the corneal injury.

The irritation of the conjunctivae consisted of readness, chemosis and discharge. Necrosis of the ocular tissue was observed in all animals from 24 hours after instillation onwards.

No signs of an early recovery of the ocular injury was observed. It was therefore decided to terminate the study and to sacrifice the animals for ethical reasons immediately after the 72 hour observation.
Other effects:
Remnants of the test substance coloration were present in the eyes of all animals on Day 1.

No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Any other information on results incl. tables

Animal # 1467 1468 1483
Time after Dosing Tissue/ Finding Score Fluor. Area (%) Comments Score Fluor. Area (%) Comments Score Fluor. Area (%) Comments
1 hour Corneal Opacity 2 D 2 D 2 D
Corneal Opacity area 2   1   1  
Iris 1   1   1  
Conj. Redness 3 (A)   3 (B)   2 (ABC)  
Conj chemosis 4 (ABC)   3 (ABC)   3 (ABC)  
Discharge 1   2   2  
24 hours Corneal Opacity 2 75 E 2 75 E 2 75 E
Corneal Opacity area 3   3   3  
Iris 1   1   1  
Conj. Redness 3 (AB)   3 (B)   3 (B)  
Conj chemosis 4 (ABC)   4 (ABC)   4 (ABC)  
Conj. Discoloration * B   B   B  
Discharge 1   2   3  
48 hours Corneal Opacity 2 E 3  - 3 F
Corneal Opacity area 3   1   1  
Iris 1   1   1  
Conj. Redness 3 (AB)   3 (ABC)   3 (ABC)  
Conj chemosis 4 (ABC)   3 (ABC)   4 (B)  
Conj. Discoloration * AB   b   B  
Discharge 2   3   3  
72 hours Corneal Opacity 2 75 - 3 75 - 3 75 F
Corneal Opacity area 3   1   2  
Iris 1   1   1  
Conj. Redness 3 (ABC)   3 (ABC)   3 (ABC)  
Conj chemosis 4 (A)   3 (ABC)   4 (A)  
Conj. Discoloration * AB   B   AB  
Discharge 2   2   2  
A = Eyelids
B = Nictitating membrane
C = Sclera
D = Remnants of the test substance in the eye
E = Approx. 25% of the cornea and iris could not be scored, due to oedema of the eyelids
F= Approx. 35% of the iris could not be scored due to corneal damage
* Discoloration = Gray/white discoloration of the indicated ocular tissues

Applicant's summary and conclusion

Interpretation of results:
other: Risk of serious damage to the eyes (R41)
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Based on the necrosis of the ocular tissues and on the degree and persistence of the corneal injury, it was concluded that ocular corrosion had occurred by instillation of Epyrrol into the rabbit eye in all three animals.
Executive summary:

Based on the necrosis of the ocular tissues and on the degree and persistence of the corneal injury, it was concluded that ocular corrosion had occurred by instillation of Epyrrol into the rabbit eye in all three animals. Epyrrol has serious risk of irreversible damage to eyes. Eppyrol therefore meets the criteria for classification under CLP as H314, and under DSD as R41.