Catalase

Eye irritation

Administrative data

Endpoint:

eye 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 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2012

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 / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Received from Robinson Services, Inc. Mocksville, NC, on July 30 and August 6, 2014.
- Sex: Female. All animals assigned to test were nulliparous and non-pregnant
- Age at study initiation: Young adult (11 weeks)
- Weight at study initiation: 2509-2644 grams
- Housing: The animals were singly 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: 7 or 12 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19-22ºC
- Humidity: 46-52%
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12 hour light/dark cycle

IN-LIFE DATES: From August 11, 2014 to August 16, 2014

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

other: The other eye of each rabbit remained untreated with the test substance and served as a control.

Amount / concentration applied:

0.1 mL

Duration of treatment / exposure:

Eyelids were held together for about one second and then released

Observation period (in vivo):

Ocular irritation was evaluated using a white light source at 1, 24, 48, and 72 hours post-instillation.

Number of animals or in vitro replicates:

3

Details on study design:

Preparation and Selection of Animals:
Prior to instillation (within 24 hours), both eyes of a group of animals were examined using a white light source and a fluorescein dye procedure. One drop of 2% ophthalmic fluorescein sodium was instilled into both eyes of each rabbit. The eyes were rinsed with physiological saline (0.9% NaCl) after instillation of the fluorescein and then evaluated for corneal damage using an ultraviolet light source. Just prior to instillation, the eyes were re-examined with a white light source and scored for abnormalities according to the "Scale for Scoring Ocular Lesions" Draize et al. Three healthy, naive animals (not previously tested) without pre-existing ocular irritation were selected for test. Initially, only one rabbit was placed on test. In the absence of any irritation in this animal, the remaining two animals were tested to confirm the result. A systemic analgesic (Buprenorphine SR®) was administered to relieve potential discomfort associated with eye irritation which provides therapeutic relief for periods of up to 76 hours. Prior to test substance instillation, 0.1 mg/kg of body weight of the analgesic was administered to the animals and at appropriate intervals to maintain therapeutic blood levels.

Instillation:
Prior to instillation, 2-3 drops of ocular anesthetic (Tetracaine Hydrochloride Ophthalmic Solution USP, 0.5%) were placed into both the treated and control eye of each animal. One-tenth of a milliliter of the test substance was then instilled into the conjunctival sac of the right eye of each rabbit by pulling the lower lid away from the eyeball. The upper and lower lids were then gently held together for about one second before releasing to minimize loss of the test substance. The other eye of each rabbit remained untreated with the test substance and served as a control. The rabbits were then returned to their designated cages.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1, 2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 2

Irritation parameter:

iris score

Basis:

animal: #1, 2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: #1, 2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 4

Irritation parameter:

chemosis score

Basis:

animal: #1, 2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

There was no ocular irritation noted in any treated eye during this study.

Other effects:

All animals appeared active and healthy and gained body weight during the study.

Any other information on results incl. tables

Table 1. INDIVIDUAL SCORES FOR OCULAR IRRITATION

	Rabbit No.: 3401 (Female)				Rabbit No.: 3402 (Female)				Rabbit No.: 3403 (Female)
	Hour				Hour				Hour
	1	24	48	72	1	24	48	72	1	24	48	72
I. Cornea
A. Opacity	0	1	0	0	0	1	0	0	0	1	0	0
B. Area	4	4	4	4	4	4	4	4	4	4	4	4
(AxB)x5	0	0	0	0	0	0	0	0	0	0	0	0
II. Iris
A. Values	0	0	0	0	0	0	0	0	0	0	0	0
Ax5	0	0	0	0	0	0	0	0	0	0	0	0
III. Conjunctivae
A. Redness	0	0	0	0	0	0	0	0	0	0	0	0
B. Chemosis	0	0	0	0	0	0	0	0	0	0	0	0
C. Discharge	0	0	0	0	0	0	0	0	0	0	0	0
(A+B+C)x2	0	0	0	0	0	0	0	0	0	0	0	0
Total	0	0	0	0	0	0	0	0	0	0	0	0

Table 2. MEAN SCORES FOR INDIVIDUAL RABBITS

Rabbit Number	Corneal Opacitya	Iritisa	Conjunctival Redness	ConjunctivalChemosis
3401	0	0	0	0
3402	0	0	0	0
3403	0	0	0	0
Calculated from the 24-, 48-, and 72-hour scores (EEC/GHS).

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, Catalase should not be classified as an eye-irritant.

Executive summary:

The study was conducted to assess the ocular irritation potential of Catalase according to the OECD Guideline No. 405 in compliance with GLP.

 

The study was conducted in a stepwise fashion. Initially, one-tenth of a milliliter of the test substance was instilled into the conjunctival sac of the right eye of one healthy rabbit by pulling the lower lid away from the eyeball. The upper and lower lids were then gently held together for about one second before releasing to minimize loss of the test substance. The left eye remained untreated and served as a control. Since there was no irritation observed in this animal, the test was completed on two additional animals. There was no ocular irritation noted in any treated eye during this study.

 

Under the test conditions, Catalase should not be classified as an eye-irritant.