Glucanase, endo-1,3(4)-β-

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10-04-1991 to 12-07-1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1992

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EEC. Acute Toxicity (Skin irritation). Section 84 (L383A/124-L383A/127) of the Annex to the European Communities Directive 92/69/EEC. The Official Journal of the European communities L383A (vol. 35).

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Active enzyme protein of Beta-glucanase, endo-1,3(4) (CAS no, EC no., EC name endo-1,3(4)-beta-glucanase, enzyme class 3.2.1.6)

Molecular formula:

Not applicable

IUPAC Name:

Active enzyme protein of Beta-glucanase, endo-1,3(4) (CAS no, EC no., EC name endo-1,3(4)-beta-glucanase, enzyme class 3.2.1.6)

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.: PPC 5569
- Expiration date of the lot/batch: December 2006
- Storage condition of test material: Frozen in the dark

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: bred by Novozymes A/S (previously Novo Nordisk A/S)
- Age at study initiation: 20-21 weeks
- Weight at study initiation: 2.9-3.1 kg
- Housing: The animals were housed individually in plastic cages with mesh floor (Scanbur).
- Diet (e.g. ad libitum): The animals were fed 130 g daily of a standard diet (Altromin 2110, Standard Diet for Rabbits).
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 6 animals

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3'C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): A light cycle of 12 hours light and 12 hours darkness was controlled automatically.

IN-LIFE DATES: From: 1996-04-16 To: 1996-05-22

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): used as is

Duration of treatment / exposure:

4 hours.

Observation period:

0.5-1, 24, 48, 72 hours after treatment. If skin reactions were observed, further assessments were conducted up to 21 days after test material application to determine the reversibility of the reactions.

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- % coverage: 100%
- Type of wrap if used: 0.5 mL of test material was applied to the intact skin on each rabbit under 2.5 cm x 2.5 cm patch of gauze. The patch was covered with 3M Micropore Surgical tape 1530-3 to make a semi occlusive bandage. Furthermore an elastic tubular support bandage was applied to prevent the rabbit from damaging the patch.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The patches were removed gently and the remaining test substance washed off with water.
- Time after start of exposure: Exposure time 4 hours

SCORING SYSTEM:
- Method of calculation:

Erythema and eschar formation
No erythema ...........................................................................................................................0
Very slight erythema (barely perceptible) ..............................................................................1
Well defined erythema ............................................................................................................2
Moderate to severe erythema ................................................................................................3
Severe erythema (beet redness) to slight eschar formations (injuries in depth) ...................4

Oedema formation
No oedema ..............................................................................................................................0
Very slight oedema (barely perceptible) .................................................................................1
Slight oedema (edges of area well defined by definite raising) ..............................................2
Moderate oedema (raised approximately 1 mm) ......................................................................3
Severe oedema (raised more than 1 mm and extending beyond area of exposure) ..............4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0.67

Max. score:

2

Reversibility:

fully reversible within: 4-5 days

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.67

Max. score:

2

Reversibility:

fully reversible within: 5 days

Remarks on result:

positive indication of irritation

Results 3

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Remarks on result:

no indication of irritation

Results 5

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

All symptoms were fully reversible within 4-5 days after exposure.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this test and criterta of the EEC Guidelines Pentopan/Biofeed Plus, batch PPC 5569 should not be classified as a skin irritant.

Executive summary:

A test for skin irritation in rabbits was performed with Pentopan/Biofeed Plus, batch PPC 5569 in accordance with OECD Guideline No. 404 and EEC Directive 1992.

0.5 mL test substance was applied once under a semi occlusive bandage to the shaved back skin of each of 3 rabbits. The length of exposure was 4 hours. The primary skin irritation was evaluated 0.5-1, 24, 48 and 72 hours after removal of the test substance. Irritation was not observed and neither were any clinical reaction to treatment recorded.

Under the test conditions employed, Pentopan/Biofeed Plus was non irritant to rabbit skin and should not be classified as a skin irritant.