4-phytase

Eye irritation

Administrative data

Endpoint:

eye 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:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Active enzyme of 4-phytase (CAS 9001-89-2, IUBMB 3.1.3.26)

Molecular formula:

Not applicable, see remarks.

IUPAC Name:

Active enzyme of 4-phytase (CAS 9001-89-2, IUBMB 3.1.3.26)

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:

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 4

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 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:

% Total Organic Solids: 25.27%

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River, D-88353, Germany
- Age at study initiation: not specified
- Weight at study initiation: 3.1 - 5.1 kg
- Housing: Kept individually in PPO- plastic cages (floor area: 4200 cm2) with perforated floor. The tray under the floor was cleaned twice per week.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 4 months for first animal and 2 weeks for additional animals

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21°C ± 3C°
- Humidity (%): 55% ± 15%
- Air changes (per hr): 10 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL of undiluted test item was used (equivalent to 26.51 mg enzyme concentrate dry matter/mL or 15.64 mg active enzyme protein/mL)

Duration of treatment / exposure:

Single exposure

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 females

Details on study design:

REMOVAL OF TEST SUBSTANCE: Test substance was not removed after application to the eye.

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: At 24 hours after treatment, instillation of Fluorescein followed by flushing with 0.9% NaCl was included. Both eyes were examined again using UV light to detect corneal damage.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: ALL

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

animal: ALL

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

chemosis score

Basis:

animal: ALL

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal: ALL

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No irritation was observed in any of the animals.

Other effects:

No clinical signs were observed in daily observations

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

No irritation was observed in any animal tested.

Executive summary:

The objective of this study was to assess the ocular irritation potential of the test substance following OECD Guideline 405.  The initial test, with 0.1 mL of test substance (equivalent to 26.51 mg enzyme concentrate dry matter/mL or 15.64 mg active enzyme protein/mL). applied to the left eye and the right eye as control, was conducted on one rabbit. After 24 hours, fluorescein was instilled and rinsed with 0.9% NaCl. The eye was examined with a UV light to detect corneal damage. The eye was also examined at 48 and 72 hours. Changes in the eye were graded according to a numerical scale 1, 24, 48, and72 hours after treatment. The confirmatory tests were carried out on an additional 2 rabbits and the tests and examinations were performed in a similar manner.

 

No clinical signs were observed in the animals during the daily observations. The mean values for all three rabbits were 0 for cornea opacity, iris lesion, chemosis of conjunctiva, and redness of conjunctiva.