Peroxidase

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1996-04-19 to 2000-03-30

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Peroxidase

EC Number:

232-668-6

EC Name:

Peroxidase

Cas Number:

9003-99-0

Molecular formula:

Not applicable, see remarks.

IUPAC Name:

Peroxidase IUBMB 1.11.1.7

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.

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.

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.

IUPAC Name:

Lipids as a constituent of enzyme deriving from the fermentation or extraction process

Test material form:

solid: particulate/powder

Details on test material:

- Lot/batch No.: PPX 5156
- Expiration date of the lot/batch: At least 2005.

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 (formerly Novo Nordisk A/S), Denmark.
- Age at study initiation: 17-19 weeks.
- Weight at study initiation: Between 2.6 - 2.7 kg.
- Housing: Individually in plastic cages with mesh floor and automatic watering.
- Diet: 130 g standard diet daily (Altromin 2110).
- Water: Ad libitum tap water.
- Acclimation period: 7 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3 °C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

physiological saline

Remarks:

0.5 g of test substance moistened with 0.9 w/v % NaCl.

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g (equivalent to 0.49 g enzyme concentrated dry matter)

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.5 g of test substance moistened with 0.9 w/v % NaCl.

Adjacent areas of untreated skin served as control.

Duration of treatment / exposure:

4 hours

Observation period:

0.5-1, 24, 48 and 72 h after patch removal using the scoring system.
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: The test compound peroxidase was assessed by semiocclusive application of 0.5 g of the test material, moisted with saline, to an area of 2.5 x 2.5 cm on the shaved skin on the back of three rabbits for 4 hrs.

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

OBSERVATION TIME POINTS
(indicate if minutes, hours or days): 0.5-1, 24, 48 and 72 h after patch removal using the scoring system. In addition, the animals were observed daily for systemic signs of toxicity.

SCORING SYSTEM:

Erythema and eschar fonnation Grade
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:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 6 days

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 5

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 9 days

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:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Dermal reaction was observed at the test site of all animals at the readings 1, 24 and 48 hours after test substance removal. The scores never exceeded 2. At 72 hours after test substance removal and the following two days two animals showed redness at the site of application. From day 6 until day 8 skin reaction (redness) was seen in only one animal. The reaction never exceeded a score of one. Nine days after removal of the test substance no reaction was observed.

Other effects:

No other effects.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Conclusions:

Under the conditions of this test and criteria of the EEC Guideline, peroxidase batch PPX 5156, was classified as 'non-irritant to skin' as the reaction never exceeded a score higher than 1 and as the skin reaction had disappeared within the first 9 days of the 21-day observation period.

Executive summary:

The acute dermal irritant effect of peroxidase was investigated according to OECD TG 404 (1992) “Acute Dermal Irritation/Corrosion”. The study was conducted in accordance with GLP.

 

The study was performed on three New Zealand White rabbits. 0.5 g test substance (equivalent to 0.49 g enzyme concentrate dry matter) 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 observed in two animals at the reading 72 hours after test substance removal. No irritation was observed nine days after test substance removal. No systemic effects were observed.

 

Under the conditions of this test and criteria of the EEC Guideline, peroxidase batch PPX 5156, was classified as 'non-irritant to skin' as the reaction never exceeded a score higher than 1 and as the skin reaction had disappeared within the 21 day observation period.