Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 June 2010 - 24 June 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP- Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material : J-37
- Substance type: red powder
- Physical state: powder
- Analytical purity: 99.3%
- Lot/batch No.: 91116
- Expiration date of the lot/batch: 16th April 2011
- Storage condition of test material: at room temperature in the dark in desiccators
- Stability under test conditions: stable

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Harlan, Belton, Leics, England.
- Age at study initiation: 6 weeks old
- Weight at study initiation: At least 1.0 kg
- Housing: animals were indvidually housed in labeled cages with perforated floors (Ebeco, Germany, dimensions 67 x 62x 55 cm) and shelters (Ebeco, Germany, dimensions 40 x 32x 23 cm).
- Diet : pelleted diet for rabbits (Global Diet 2030 from Harlan Teklad, Mucedola, Milanese, Italy) approximately 100 grams per day. Hay (TecniLab- BMI BV, Someren, The Netherlands) was provided at least three times a week.
- Water : free access to tap water.
- Acclimation period: at least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 (Actual range: 19.0 - 23.4)
- Humidity (%): 40 - 70 (Actual range: 38 - 81)
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours artifical fluorescent light and 12 hours darkness per day.

IN-LIFE DATES: From: 14th June 2010 To: 24th June 2010

Test system

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): animals were treated by instillation of, on average, 32.3 mg (range 32.2 -32.5) of the test substance ( a volume of approximately 0.1 mL) as the maximum required amount in the conjuctival sac of one of the eyes after gently pulling the lower lid away from the eyeball.

Duration of treatment / exposure:
4 hours
Observation period (in vivo):
The eye of each animal were examined approximately 1, 24, 48 and 72 hours after instillation of the test substance.
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): No.

SCORING SYSTEM: accoridng to Draize (1944).

TOOL USED TO ASSESS SCORE: fluorescein. Immediately after the 24- hour observation, a solution of 2% fluorescein (Merck, Darmstadt, Germany) in water (adjusted tp pH 7.0) was instilled into both eyes of each animal to quantitavely determine corneal epithelial damage. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24, 48 & 72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24, 48 & 72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 24, 48 & 72 h
Score:
0.6
Max. score:
3
Reversibility:
fully reversible within: 72 h
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 24, 48 & 72 h
Score:
0
Max. score:
4

Any other information on results incl. tables

Instillation of approximately 32 mg of the test substance (a volume of approximately 0.1 mL) into one eye of each of three rabbits resulted in effects on the iris and conjunctivae. Iridial irritation grade 1 was observed in all animals and had resolved within 24 hours. The irritation of the conjunctivae consisted of redness, chemosis and discharge and completely resolved within 48 hours in one animal and within 72 hours in the other two animals (Table 1).

Treatment of the eyes with 2% fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage.

Remnants of the test substance were present in the eye on Day 1 and on the outside of the eyelids throughout the observation period. Red/purple staining of the fur on the head and paws, caused by the test substance, was noted throughout the observation period.

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

 

Table 1. Individual eye irritation scores

Animal

Time after dosing

Cornea

Iris

Conjunctivae

Comments

Opacity (0-4)

Area (0-4)

Fluor area (%)2

(0-2)

Redness (0-3)

Chemosis (0-4)

Discharge (0-3)

Comments

 

4341

1 Hour

0

0

-

1

2

1

2

b,e,c

24 Hour

0

0

0

0

1

0

0

e,c

48 Hour

0

0

-

0

1

0

0

e,c

72 Hour

0

0

-

0

0

0

0

e,c

440

1 Hour

0

0

-

1

1

1

1

b,e,c

24 Hour

0

0

0

0

1

0

1

e

48 Hour

0

0

-

0

0

0

0

e

72 Hour

0

0

-

0

0

0

0

e

442

1 Hour

0

0

-

1

2

1

1

b,e,c

24 Hour

0

0

0

0

1

0

1

e

48 Hour

0

0

-

0

1

0

0

e

72 Hour

0

0

-

0

0

0

0

e

 1Sentinel,2Green staining after fluorescein treatment (percentage of total corneal area)

Comments:

b: Remnants of the test substance in the eye, c: Remnants of the test substance on the outside of the eyelids and e: Red/ purple staining of the head and paws by the test substance.

Table 2. Mean value eye irritation scores.

Animal

Mean 24, 48 and 82 Hours

Corneal opacity

Iris

Conjunctivae

Redness

Chemosis

434

0.0

0.0

0.7

0.0

440

0.0

0.0

0.3

0.0

442

0.0

0.0

0.7

0.0

 

 

 

Table 3: Animal Specifications

Animal

Sex

Age at start (weeks)

Body weight (grams)

Prior to application

At termination

434

M

12

2525

2601

440

M

7-9

1643

1749

442

M

7-9

1543

1636

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU