(7R)-7-(4-carboxybutanamido)cephalosporanate amidohydrolase (enzyme substance)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 13-JUN-2002 to 01-JUL-2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The test was performed according to OECD guideline and GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

Deviations from the maximum level of temperature (with a maximum of 1°C) and from the maximum level for relative humidity (up to 23%) occurred . These protocol deviations were considered not to have affected the study integrity.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

deoxyribose-phosphate aldolase EC 4.1.2.4.

Cas Number:

9026-97-5

Molecular formula:

Not applicable, please see remarks

IUPAC Name:

deoxyribose-phosphate aldolase EC 4.1.2.4.

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:

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 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:

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

Remarks:

Tests based on dissolved material in the concentrate

Details on test material:

- Name of test material (as cited in study report): Aldolase (IUB 4.1.2.4)
- Substance type: enzyme
- Physical state: liquid
- Stability under test conditions: data not available
- Storage condition of test material: In freezer in the dark

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan, Horst, The Netherlands
- Age at study initiation: at least 6 weeks old
- Weight at study initiation: 1309g, 1499g and 2367g
- Housing: individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56x44x37.5cm )
- Diet: Standard laboratory rabbit diet (Teklad Global Rabbit Diet, code 2030, Harlan, Blackthorn, England) approx. 100 g. per day
- Water: Free access to tap-water
- Acclimation period: at least 5 days before start of treatment under laboratory conditions


ENVIRONMENTAL CONDITIONS
- Temperature: 21 +/- 3°C
- Humidity: 30-70%
- Air changes: approximately 15 air changes per hour
- Photoperiod: 12 hours artificial fluorescent light and 12 hours dark per day


IN-LIFE DATES: data not available

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The other eye remained untreated and served as the reference control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 mL
- Concentration: undiluted

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 male animals

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no

SCORING SYSTEM: according to OECD and EU guidelines


TOOL USED TO ASSESS SCORE: Immediately after the 24 hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24 - 72h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Remarks:

no effect observed

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24 - 72h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Remarks:

no effect observed

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 24 - 72h

Score:

0

Max. score:

3

Reversibility:

fully reversible within: 24h

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24 - 72h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Remarks:

no effect observed

Irritant / corrosive response data:

Irritation:
Instillation of 0.1 ml of the test substance into one eye of each of three rabbits resulted in irritation of the conjunctivae in 2 animals, which was seen as redness. The irritation had completely resolved within 24 hours in both animals.
No iridial irritation or corneal opacity was observed, and treatment of the eyes with 2% fluorescein, 24 hours after test substance instillation revealed no corneal epithelial damage in any of the animals.

Corrosion:
There was no evidence of ocular corrosion.

Other effects:

Colouration:
No staining of (peri) ocular tissues by the test substance was observed.

Toxicity / Mortality:
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Any other information on results incl. tables

Table 1: Irritant/corrosive response data for each animal at each observation time up to removal of each animal from the test

Score at time point / Reversibility	Cornea	Iris	Conjunctivae	Chemosis
	Max. score: 4	Max. score: 2	Max. score: 3	Max. score: 4
60 min	0 / 0 / 0	0 / 0 / 0	1 / 1 / 0	0 / 0 / 0
24 h	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0
48 h	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0
72 h	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0
Average 24h, 48h, 72h	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0
Maximum average score (including area affected, max 110)	0	0	0	0
Reversibility	-	-	yes	-
Average time (h) for reversion	-	-	24h	-

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

ALDOLASE (I.U.B. 4.1.2.4) does not have to be classified and has no obligatory labelling requirement for eye irritation according to EU criteria.

Executive summary:

The study was carried out based on the guidelines described in: EC Commission Directive 92/69/EEC, 8.5, "Acute Toxicity - Eye irritation" and OECD No.405, "Acute Eye Irritation/Corrosion".

Single samples of 0.1 ml of ALDOLASE (I.U.B. 4.1.2.4) were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours after instillation.

Instillation of the test substance resulted in irritation of the conjunctivae in 2 animals, which was seen as redness. The irritation had completely resolved within 24 hours in both animals. No iridial irritation or corneal opacity was observed, and treatment of the eyes with 2% fluorescein, 24 hours after test substance instillation revealed no corneal epithelial damage in any of the animals.

Based on these results and according to the EC criteria for classification and labelling requirements for dangerous substances and preparations (Council Directive 67/548/EEC), ALDOLASE (I.U.B. 4.1.2.4) does not have to be classified and has no obligatory labelling requirement for eye irritation.