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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (GLP, QUA)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report Date:
2000

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)
Qualifier:
according to
Guideline:
other: US FDA Title 21 Code of Federal Regulations Part 58; US EPA (FIFRA), Title 40 Code of Federal Regulations Part 160; US EPA (TSCA), Title 40 Code of Federal Regulations Part 792;
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries, 59 NohSan, Notifications No.3850; Japanese Ministry of International Trade and Industry, Kanpogyo No.39 Environmental Agency, Kikyoku No.85; Japanese Ministry of Health and Welfare, Ordinance No.21
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
- Name of test material (as cited in study report): FR-6120
- Physical state: white powder
- Analytical purity: ca. 99.5%
- Impurities (identity and concentrations): water (0.27%), Melamine (0.19%) and cyanuric acid (0.012%) (weight%)
- Purity test date: 04 May 2000
- Lot/batch No.: 37432-18-3
- Expiration date of the lot/batch: 22 May 2001
- Stability under test conditions: not indicated
- Storage condition of test material: 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: Albino Rabbit, New Zealand White, (SPF-Quality) from Charles River Deutschland, Kisslegg, Germany.
- Age at study initiation: 6 weeks
- Weight at study initiation: less than 3.5 kg (mean bw: 1.732 kg)
- Housing: Individually housed in labelled cages with perforated floors (Scanbur, Denmark) and equipped with an automatic drinking system (ITL, Bergen, The Netherlands)
- Diet (e.g. ad libitum): standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Germany) approx. 100 g. per day.
- Water (e.g. ad libitum): free access to tap-water.
- Acclimation period: at least 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
A controlled environment was maintained in the room with optimal conditions
- Temperature (°C): 21°C
- Humidity (%): 50%
- Air changes (per hr): ca. 15
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours dark per day

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 (volume or weight with unit): each animal was treated by instillation of 37.9 ± 0.5 mg of the test substance (a volume of approximately 0.1 ml) in the conjunctival sac of one of the eyes 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.
Duration of treatment / exposure:
the test substance was not washed out
Observation period (in vivo):
up to 72 hours; the eyes of each animal were examined approximately 1, 24, 48 and 72 hours after instillation of the test substance. The irritation scores and a description of all other (local) effects were recorded.
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): the test substance was not washed out

SCORING SYSTEM: the irritation was assessed according to the OECD numerical scoring system. The results were evaluated according to the EC criteria for classification and labeling requirements for dangerous substances and preparations.

TOOL USED TO ASSESS SCORE: in case standard lighting was considered inadequate for observing minor effects, eye examinations were performed using an ophthalmic examination lamp.
In cases of equivocal results when comparing the treated and untreated eyes, the illustrated guide from the Consumer Product Safety Commission, Washington, D.C. 20207 was used for additional control purposes.

OTHER: immediately after the 24 hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively 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
Remarks:
coneal opacity
Basis:
mean
Remarks:
of all tested animals
Time point:
other: 24-48-72-hour
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
other: see Table 1
Irritation parameter:
iris score
Basis:
mean
Remarks:
of all tested animals
Time point:
other: 24-48-72-hour
Score:
0
Max. score:
2
Reversibility:
fully reversible within: 24 hours (iridial irritation was observed at the 1-hour observation period)
Remarks on result:
other: see Table 1
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
of all tested animals
Time point:
other: 24-48-72-hour
Score:
0.3
Max. score:
3
Reversibility:
fully reversible
Remarks:
48 hours
Remarks on result:
other: see Table 1
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
of all tested animals
Time point:
other: 24-48-72-hour
Score:
0
Max. score:
4
Reversibility:
fully reversible
Remarks:
24 hours (chemosis was observed at the 1-hour observation period)
Remarks on result:
other: see Table 1
Irritant / corrosive response data:
- The instillation 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 (see Table 1);
- Iridial irritation (grade 1) was observed in all animals after 1 hour only;
- Irritation of the conjunctivae was seen as redness, chemosis and discharge, which had completely resolved within 48 hours in all animals;
- No corneal opacity was observed, and treatment of the eyes with 2% fluorescein, 24 hours after test substance instillation revealed no corneal epithelial damage in any of the animals.
Other effects:
- Coloration: remnants of the test substance were present in the eyes of all animals on day 1.
- Toxicity / Mortality: 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

Table 1: Individual eye irritation scores

Animal#

Time after dosing

Cornea

Iris

Conjunctivae

Opacity

Area

Fluor area (%)

Redness

Chemosis

Discharge

Comments

 

First animal (#91)

1 hour

0

0

 

1

2

2

1

b

24 hours

0

0

0

0

1

0

0

-

48 hours

0

0

 

0

0

0

0

-

72 hours

0

0

 

0

0

0

0

-

 

Second animal (#93)

1 hour

0

0

 

1

2

2

1

b

24 hours

0

0

0

0

1

0

0

-

48 hours

0

0

 

0

0

0

0

-

72 hours

0

0

 

0

0

0

0

-

 

Third animal (#96)

1 hour

0

0

 

1

2

2

1

b

24 hours

0

0

0

0

1

0

0

-

48 hours

0

0

 

0

0

0

0

-

72 hours

0

0

 

0

0

0

0

-

Fluor area (%): green staining (percentage of total corneal area) after fluorescein treatment; b: Remnants of the test substance in the eye.

Applicant's summary and conclusion

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