Catalase

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 21, 2014 to October 24, 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Active enzyme protein of catalase (EC no. 232-577-1, CAS no. 9001-05-2, EC name Catalase, Enzyme class no. 1.11.1.6)

Molecular formula:

Not applicable, see remarks

IUPAC Name:

Active enzyme protein of catalase (EC no. 232-577-1, CAS no. 9001-05-2, EC name Catalase, Enzyme class no. 1.11.1.6)

Constituent 2

Reference substance name:

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

Molecular formula:

Not available. See remarks.

IUPAC Name:

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

Constituent 3

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

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:

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

Test material form:

liquid

Details on test material:

- Substance type: UVCB
- Physical state: light brown liquid
- Stability under test conditions: The neat test substance was demonstrated to be stable under the conditions of the study.

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

SOURCE ANIMAL
- Source: Received from Robinson Services, Inc. Mocksville, NC, on July 23 and 30, 2014
- Sex: Female. The animals assigned to test were nulliparous and non-pregnant.
- Age at study initiation (in days): Young adult. (10-11 weeks)
- Weight at study initiation: 2524-2818 grams
- Housing: The animals were individually housed in suspended stainless steel perforated bottom caging, which conform to the size recommendations in the most recent Guide for the Care and Use of Laboratory Animals. Enrichment (e.g., toy) was placed in each cage. Litter paper was placed beneath the cage and was changed at least three times per week.
- Diet (e.g. ad libitum): Harlan Teklad Global High Fiber Rabbit Diet® 2031. A designated amount of the diet (approximately 150 grams/day) and a Premium Timothy CubeTM (Ontario Dehy Inc.) were available to each rabbit.
- Water (e.g. ad libitum): Filtered tap water was supplied ad libitum.
- Acclimation period: 8 or 13 days

ENVIRONMENTAL CONDITIONS
- Temperature: 18-22°C
- Humidity: 46-60%,
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12 hour light/dark cycle

IN-LIFE DATES: From August 5, 2014 to August 10, 2014

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.5 mL

Duration of treatment / exposure:

4 hours

Observation period:

Individual dose sites were scored according to the Draize scoring system immediately following patch removal (initial rabbit only) and at approximately 30-60 minutes, 24, 48, and 72 hours after patch removal.

Number of animals:

3

Details on study design:

Preparation and Selection of Animals:
Approximately 24 hours prior to application, the animals were prepared by clipping the dorsal area and the trunk. On the day of dosing, but prior to application, the animals were examined for health and the skin checked for any abnormalities. Three healthy naive animals (not previously tested) without pre-existing skin irritation were selected for test. Initially, only one rabbit was placed on test. Since there was only slight dermal irritation noted at the dose site of this animal at 24 hours after patch removal, the test was completed with two additional rabbits. A systemic analgesic (Buprenorphine SR®) was administered to relieve potential discomfort associated with skin irritation which provides therapeutic relief for periods of up to 76 hours. Prior to test substance application, 0.1 mg/kg of body weight of the analgesic was administered to the animals and at appropriate intervals to maintain therapeutic blood levels.

Application of Test Substance:
Five-tenths of a milliliter of the test substance was applied to one 6-cm2 intact dose site on each animal and covered with a 1-inch x 1-inch, 4-ply gauze pad. The pad and entire trunk of each animal were then wrapped with semi-occlusive 3-inch Micropore tape to avoid dislocation of the pad. Elizabethan collars were placed on each rabbit and they were returned to their designated cages. No other substances were tested on these rabbits. After 4 hours of exposure to the test substance, the pads and collars were removed and the test sites were gently cleansed with a 3% soap solution followed by tap water and a clean paper towel to remove any residual test substance.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal: #1, 3

Time point:

24/48/72 h

Score:

0.7

Max. score:

2

Reversibility:

fully reversible

Remarks:

72 hours

Results 2

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 3

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.7

Max. score:

1

Reversibility:

fully reversible within:

Remarks:

48 hours

Results 4

Irritation parameter:

edema score

Basis:

animal: #2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

1

Irritant / corrosive response data:

Erythema (scores of 1 and 2) was noted for two of three treated sites and edema (score of 1) was noted at one treated site at the 30-60 minute scoring interval. Erythema and edema cleared from the dose sites by 72 hours.

Other effects:

All animals appeared active and healthy during the study. One rabbit lost body weight during the study. There were no other clinical signs observed

Any other information on results incl. tables

Table 1. INDIVIDUAL SKIN IRRITATION SCORES ( ERYTHEMA/EDEMA)

Animal No.	Sex	Time After Patch Removal
		Immediately after patch removal	30-60 mins	24 hrs	48 hrs	72 hrs
3501	F	2/1	2/1	1/0	1/0	0/0
3502	F	N/A	0/0	0/0	0/0	0/0
3503	F	N/A	1/0	1/0	1/0	0/0

N/A – Not Applicable

Table 2: SUMMARY OF MEAN SCORES FOR DERMAL RESPONSES

RABBIT NUMBER	ERYTHEMA	EDEMA
3501	0,7	0
3502	0	0
3503	0,7	0
Calculated from the 24-, 48-, and 72-hour dermal responses(EEC/GHS).

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, Catalase should not be classified as a skin-irritant.

Executive summary:

A primary skin irritation test was conducted with rabbits to determine the potential of Catalase to produce irritation after a single topical application. This study was conducted according to the OECD Guideline 404 in compliance with the GLP.

 

The study was conducted in a stepwise manner using a single patch applied initially to one rabbit for 4 hours. Five-tenths of a milliliter of the test substance was applied to the skin of one healthy rabbit for 4 hours. The dose site was evaluated for dermal irritation immediately after patch removal and at 30-60 minutes and 24, 48, and 72 hours after patch removal. Since there was only slight dermal irritation noted at the dose site of this animal at 24 hours after patch removal, the test was completed on two additional animals, as described above, except the dose sites were not evaluated immediately after patch removal. Erythema (scores of 1 and 2) was noted for two of three treated sites and edema (score of 1) was noted at one treated site at the 30-60 minute scoring interval. Erythema and edema cleared from the dose sites by 72 hours.

Under the study conditions, Catalase should not be classified as a skin-irritant.