Mucorpepsin

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1982-04-20 to 1982-06-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test method was proposed by OECD Guidelines for Testing Chemicals and evaluation made according to the Code of Federal Regulations, title 16, §1500.42.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1982

Report date:

1982

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD Guidelines for Testing Chemicals and evaluation made according to the Code of Federal Regulations, title 16, §1500.42.

Deviations:

no

GLP compliance:

yes

Remarks:

GLP equivalent at the time where the conduct of the study was subject to appropriate inspection by the quality assurance unit.

Test material

Test material information

Constituent 1

Reference substance name:

Active enzyme protein of mucorpepsin (CAS no.148465-73-0, EC no 642-981-3, enzyme class 3.4.23.23)

Molecular formula:

Not applicable

IUPAC Name:

Active enzyme protein of mucorpepsin (CAS no.148465-73-0, EC no 642-981-3, enzyme class 3.4.23.23)

Constituent 2

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 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:

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 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:

solid: particulate/powder

Details on test material:

- Lot/batch No.: PPR 1289
- Expiration date of the lot/batch: No specific expiry date. At least 10 years or as long as enzyme activity is preserved

Test animals / tissue source

Species:

rabbit

Strain:

other: Danish Landstrain

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Bred by Novozymes A/S (previously Novo Industri A/S)
- Weight at study initiation: 2.9-3.2 kg
- Housing: Individually in galvanized steel cages with mesh floors and automatic watering
- Diet (e.g. ad libitum): Standard diet (Ewos Brood Stock Feed for Rabbits and Guinea Pigs).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 22 °C
- Humidity (%): 30 - 35%

IN-LIFE DATES: From: 1982-04-20 To: 1982-04-23

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

Test batch used as is:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL into the left eye

Duration of treatment / exposure:

0.1 mL of the test substance was installed into the left eye of each rabbit by gently pulling the lower lid away from the eye ball, and the test substance was carefully introduced into the resulting cup. The lids were then gently held together for one second.
The right eye, remaining untreated, served as a control.

Observation period (in vivo):

The eyes were examined at 1, 24, 48 and 72 hours after treatment.

Number of animals or in vitro replicates:

6 rabbits (3 female + 3 male)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: No
SCORING SYSTEM: The ocular reactions were recorded at 1, 24, 48 and 72 hours after instillation according to the Code of Federal Regulations, title 16 §1500.42

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 2

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 3

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal: #1, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 4

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible

Results 5

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 6

Irritation parameter:

conjunctivae score

Remarks:

Ulceration

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

No corneal opacity or iris reactions were noted at any time during the test. Mild redness and slight chemosis were seen in all of the rabbits estimated at 1 hour.
At 24 hours mild redness was seen in 1 rabbit.
Both observed reactions were fully reversible and at 48 and 72 hours no conjunctival reactions were seen in any of the rabbits.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Not classified, Criteria used for interpretation of results: OECD GHS

Conclusions:

In conclusion, Rennilase SP 252 was regarded as negative to potential for eye injury.

Executive summary:

In accordance with the proposed test method by OECD Guidelines for Testing Chemicals and with the evaluation conducted according to the Code of Fereral Regulation, title 1500.42, a test for eye irritation in rabbits was performed with rennilase SP 252, batch PPR 1289.

0.1 mL of rennilase SP 252, batch PPR 1289 was placed in the conjunctival sac of the left eye of six rabbits by pulling the lower eyelid away from the eyeball. The lids were gently held together for about one second after instillation. The untreated eye served as control. The ocular reactions were recorded at 1, 24, 48 and 72 hours after instillation according to the specified guideline.

No corneal opacity or iris reactions were noted at any time during the test. Mild redness and slight chemosis were seen in all of the rabbits estimated at 1 hour. At 24 hours mild redness was seen in 1 rabbit. At 48 and 72 hours no conjunctival reactions were seen in any of the rabbits.

In conclusion, rennilase SP 252 was classified as a 'non-irritant' to the rabbit eye.