Lyase, pectate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 1999-02-15 to 1999-05-05

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

1992

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EEC

Version / remarks:

EEC (1992). Acute Toxicity (Eye irritation). Section B5 (L383A/127-L383A/130) of the Annex to
the European Communities Commision Directive 92/69/EEC. The Official Journal of the European
Communities L.383A ( Vol. 35), 29 December 1992 (ISBN 0378-6978).

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Lyase, pectate

EC Number:

232-766-9

EC Name:

Lyase, pectate

Cas Number:

9015-75-2

Molecular formula:

Not available

IUPAC Name:

Pectate lyase IUBMB 4.2.2.2

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:

liquid

Details on test material:

- Lot/batch No.: PPE 6345
- Expiration date of the lot/batch: 2008-11-30
- Stability under test conditions: Stability at ambient temperature >4 hours
- Storage condition of test material: Stored below minus 18 degrees of C in the dark.

Test animals / tissue source

Species:

rabbit

Strain:

other: Danish Landrace, albino

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Bred by Statens Seruminstitut "Hvidesten", Denmark
- Weight at study initiation: between 2.14 - 2.56 kg
- Age at study initiation: Young adults
- Housing: Individually, in animal room with control of temperature (20±3°C) and humidity (30-70%)
- Diet: 110 g standard diet per day (Altromin 2110, Standard diet for rabbits) ad libitum
- Water: Tap water ad libitum
- Acclimation period: 14 days
- Photoperiod (hrs dark / hrs light): 12 hrs/12hrs

IN-LIFE DATES:
From: 1999-02-22 To: 1999-03-12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

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

Duration of treatment / exposure:

The test material was applied to the conjunctival sac. The lids were gently held together for approximately 1 second. The untreated eye served as a control.

Observation period (in vivo):

The reactions were judged 1, 24, 48 and 72 hours after instillation of the test material.

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: Not specified
SCORING SYSTEM: The ocular reactions were recorded at 1, 24, 48 and 72 hours after instillation according to the guideline.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal: #2, #4

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 2

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #5

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible

Results 3

Irritation parameter:

chemosis score

Basis:

animal: #2, #4, #5

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 4

Irritation parameter:

cornea opacity score

Basis:

animal: #2, #4, #5

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 5

Irritation parameter:

iris score

Basis:

animal: #2, #4, #5

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 6

Irritation parameter:

other: Tear secretion

Basis:

animal: #2, #4, #5

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

No corneal opacity, iris reactions or tear secretion were noted at any time during the test. Conjunctiva! reaction (redness, score 1) was observed in two rabbits at the 1 hour reading and in one rabbit at the 24 hour reading. Chemosis (score 1) was observed at the 1 hour reading in two animals. All signs were reversible, and no reactions were seen after 48 or 72 hours. The rabbits showed no signs of systemic reaction to the treatment.

Other effects:

None

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

No corneal opacity, iris reactions or tear secretion were noted at any time during the test. Slight conjunctival reaction and chemosis were observed earlier on. However, all signs were reversible, and no reactions were seen after 48 or 72 hours. The rabbits showed no signs of systemic reaction to the treatment.
In conclusion, pectate lyase can be classified as a ‘non irritant’ to the eye.

Executive summary:

In accordance with the OECD guideline 405, a test for eye irritation in rabbits was performed with pectate lyase, batch PPE 6345.

The eyes of three animals were checked for irritation before instillation and only healthy animals were used. 0.5 mL of pectate lyase, batch PPE 6345 was placed in the conjunctival sac of the left eye 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, iris reactions or tear secretion were noted at any time during the test. Conjunctival reaction (redness, score 1) was observed in two rabbits at the 1 hour reading and in one rabbit at the 24 hour reading. Chemosis (score 1) was observed at the 1 hour reading in two animals. All signs were reversible, and no reactions were seen after 48 or 72 hours. The rabbits showed no signs of systemic reaction to the treatment.

In conclusion, pectate lyase can be classified as a 'non-irritant' to the rabbit eye.