Proteinase, Aspergillus acid

Skin irritation / corrosion

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Test guideline 2

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

Test material

Test material information

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

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

Test system

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.

Results and discussion

In vivo

Results

Results 1

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

Results 2

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

Results 3

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

Results 4

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

Results 5

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

Results 6

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.

Applicant's summary and conclusion

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.