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Description of key information

Aspergillopepsin I was tested for skin and eye irritation in rabbits and and no dermal or eye irritation was observed.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin 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
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EEC Directive Annex I, II, III, and IV, Official Journal of the European Communities published in L 110 A, volume 36, 4 May 1993 § 3.2.6. I and § 3.2.5
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland GmbH, D-88353 Kisslegg, Germany
- Age at study initiation: not specified
- Weight at study initiation: 3.2 - 3.6 kg
- Housing: The animals were kept in a PPO-plast cage (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: approximately 2 months

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21°C ± 3°C
- Humidity (%): 55% ± 15%
- Air changes (per hr): 10/hr
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
Duration of treatment / exposure:
Initial Test: 3 minutes, 1 hour, 4 hours
Confirmatory Tests: 4 hours
Observation period:
After each patch removal (3 minutes, 1, and 4 hours) and at 1, 24, 48, and 72 after termination of exposure
Number of animals:
Initial Test: 1 female
Confirmatory Test: 2 females
Details on study design:
TEST SITE
- Area of exposure: 2.5 x 5 cm
- Type of wrap if used: Each gauze patch was secured with adhesive tape and thereafter the circumferential area of the patches was fixed with Scanpore tape. Nobafix gauze was wrapped around the trunk to secure the dressing.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): lukewarm water and mild soap

OBSERVATION TIME POINTS
- After patch removal (3 minutes and 1 and 4 hours) and 1, 24, 48, and 72 hours after termination of exposure

SCORING SYSTEM:
- Method of calculation: The scores for erythema and eschar formation and oedema formation for the 3 readings at 24, 48, and 72 hours were obtained and divided by 3. The results are the average scores for erythema and eschar formation and oedema formation.
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
Initial Test: No reactions were observed at any test site or negative control site at any of the time points.

Confirmatory Test: Not reactions were observed at any test site or negative control site at any of the time points in one rabbit. In the second rabbit, no reactions were observed at the 3-minute or 1-hour application sites. Very slight erythema was observed at the 4-hour application site one hour after termination of exposure. No other reactions were observed at any other test site or negative control site at any of the time points.
Other effects:
No clinical signs were observed in the animals during the daily observations.
Interpretation of results:
GHS criteria not met
Conclusions:
The mean score for erythema was 0.0 and the mean score for oedema was 0.0. The test substance was not irritating to the skin.
Executive summary:

The acute dermal irritant effect of the test substance was carried out according to OECD Guideline 404. No clinical signs were observed in the animals during the daily observations, except for the skin reactions observed at the test sites. The study was initiated with one rabbit, and as no skin reactions were observed in this animal at any of the observation time points, two additional animals were included in the study. Very slight erythema was observed in one animal at the 4-hour application site one hour after termination of exposure. No other reactions were observed at any test site or negative control site in any of the animals at any time points during the study. The mean score for erythema was 0.0 and the mean score for oedema was 0.0.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
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
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
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
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.
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.
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.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

The skin and eye irritation potential of Aspergillopepsin I has been tested according to OECD guidelines and in compliance with GLP. The conclusion was that it did not exert any skin and eye irritation.