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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:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report date:
2006

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
One animal used in previous eye irritation study as control. Deviation did not affect the outcome of the study as a 17-day washout period allowed prior to inclusion on this study. Also, similar reactions observed in all treated animals during the study.
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
One animal used in previous eye irritation study as control. Deviation did not affect the outcome of the study as a 17-day washout period allowed prior to inclusion on this study. Also, similar reactions observed in all treated animals during the study.
GLP compliance:
yes

Test material

Constituent 1
Reference substance name:
Active enzyme protein of Aspergillopepsin I (EC no. 232-796-2, CAS no. 9025-49-4, EC name: Proteinase, Aspergillus acid, Enzyme Class no. 3.4.23.18)
Molecular formula:
Not applicable, see remarks
IUPAC Name:
Active enzyme protein of Aspergillopepsin I (EC no. 232-796-2, CAS no. 9025-49-4, EC name: Proteinase, Aspergillus acid, Enzyme Class no. 3.4.23.18)
Constituent 2
Reference substance name:
Protein as a constituent of enzyme deriving from the fermentation or extraction process
Molecular formula:
Not available
IUPAC Name:
Protein as a constituent of enzyme deriving from the fermentation or extraction process
Constituent 3
Reference substance name:
Inorganic salts as a constituent of enzyme deriving from the fermentation or extraction process
Molecular formula:
Not available. See remarks.
IUPAC Name:
Inorganic salts as a constituent of enzyme deriving from the fermentation or extraction process
Constituent 4
Reference substance name:
Carbohydrates constituent of enzyme deriving from the fermentation or extraction process.
Molecular formula:
Not available. See remarks.
IUPAC Name:
Carbohydrates constituent of enzyme deriving from the fermentation or extraction process.
Constituent 5
Reference substance name:
Lipids as a constituent of enzyme deriving from the fermentation or extraction process
Molecular formula:
Not available. See remarks.
IUPAC Name:
Lipids as a constituent of enzyme deriving from the fermentation or extraction process
Test material form:
liquid
Details on test material:
- Lot/batch No.: AFP501K1

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland GmbH, D-88353 Kisslegg, Germany
- Age at study initiation: not specified
- Weight at study initiation: 2.7 - 3.8 kg
- Housing: The animals were kept individually in PPO-plast cages (Noryl®, floor area: 4200 cm2) with perforated floor. The tray under the cage was cleaned 2 times a week.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 46 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21°C ± 3°C
- Humidity (%): 55% ± 15%
- Air changes (per hr): 10/hour
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
Duration of treatment / exposure:
Single application
Observation period (in vivo):
1, 24, 48, and 72 hours
Number of animals or in vitro replicates:
3 females
Details on study design:
REMOVAL OF TEST SUBSTANCE: No

TOOL USED TO ASSESS SCORE: Fluorescein stain was used after the 24-hour observation.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal: mean of 3 animals
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal: mean of 3 animals
Time point:
24/48/72 h
Score:
0
Max. score:
2
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal: mean of 3 animals
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal: mean of 3 animals
Time point:
24/48/72 h
Score:
0.1
Max. score:
3
Reversibility:
fully reversible
Irritant / corrosive response data:
At the examination 1 hour after exposure, conjunctiva appeared red (some blood vessels were definitely hyperaemic – equal to score 1) in the initial animal. This reaction persisted the following day (24 hours after exposure). However, it was considered ethically acceptable to proceed with 2 additional animals as only very mild reactions had been observed in the initial animal. Similar scoring of conjunctival redness was observed in the 2 additional animals at the examination 1 hour after exposure. At all other examinations, all animals appeared normal.
Other effects:
No clinical signs were observed during the daily observations.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Only slight signs of irritation were observed at 1 and 24 hours in one rabbit. All eyes were normal by 48 hours.
Executive summary:

The eye irritant effect of the test substance was investigated according to the method OECD Guideline 405. Three female albino rabbits were used in the study. Initially, one animal was exposed to 0.1 mL of the test substance in the left eye. The right eye remained untreated and served as control. Both eyes of all animals were examined and the changes were graded according to a numerical scale 1, 24, 48, and 72 hours after treatment.

 

No clinical signs were observed in the animals during the daily observations. At the examination 1 hour after exposure, conjunctiva appeared red (some blood vessels were definitely hyperaemic – equal to score 1) in the initial animals. The reaction persisted the following day (24 hours after exposure). However, it was considered ethically acceptable to proceed with two additional animals as only very mild reactions had been observed in the initial animal. Similar scoring of conjunctival redness was observed in the two additional animals at the examination 1 hour after exposure. At all examinations, all animals appeared normal. Mean values, based on the results from the 24, 48, and 72 hour readings were calculated. The mean values were 0.0, 0.0, 0.0, and 0.1 for cornea opacity, iris lesion, chemosis of conjunctiva, and redness of conjunctiva, respectively.