cyclomaltodextrin glucanotransferase

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Remarks:

in vitro

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07-09-2007 to 13-11-2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Branching enzyme and cyclomaltodextrin glucanotransferase belong to the same enzyme sub-subclass and based on the similarity of the two enzymes, read-across is considered fully applicable.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

Adopted on 24 April 2002.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Active enzyme protein of branching enzyme (CAS no.9001-97-2 , EC name 1,4-alpha-glucan branching enzyme , Enzyme class no. 2.4.1.18)

Molecular formula:

n.a.

IUPAC Name:

Active enzyme protein of branching enzyme (CAS no.9001-97-2 , EC name 1,4-alpha-glucan branching enzyme , Enzyme class no. 2.4.1.18)

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. 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

Details on test material:

- Lot/batch No.: PPY27209
- Expiration date of the lot/batch: June 2017

Test animals / tissue source

Species:

rabbit

Strain:

other: SPF bred New Zealand White albino rabbits

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River, Germany
- Age at study initiation: males, young adult
- Weight at study initiation: 1993, 1911 and 1886 g
- Housing: Individually in stainless steel cages with perforated floor.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): An amount of ca 0.1 mL of the test substance was instilled in the conjunctiva! cul-de-sac of the right eye.
- Concentration (if solution): undiluted test sample, 7.3% TOS.

Duration of treatment / exposure:

After administration, the upper and lower eye lid were carefully closed and subsequently held together for at least one second before releasing, to prevent loss of material.

Observation period (in vivo):

1, 24, 48, 72 h after treatment.

Number of animals or in vitro replicates:

3

Details on study design:

Only rabbits without observable eye defects were used. The study was carried out with three rabbits (started with one rabbit and continued with two rabbits ca 1 hour later) and each rabbit was treated as follows: An amount of ca 0.1 mL of the test substance was instilled in the conjunctiva! cul-de-sac of the right eye. After administration, the upper and lower eye lid were carefully closed and subsequently held together for at least one second before releasing, to prevent loss of material. The other eye remaining untreated, served as a control. The reactions of the test eyes were judged at circa 1, 24, 48, 72 h after treatment

SCORING SYSTEM:

CORNEA
Opacity-degree of density (area most dense taken for reading)
No opacity ............................................................................................................................ 0
Scattered or diffuse areas, details of iris clearly visible ..................................................... 1
Easily discernible translucent area, details of iris slightly obscured ................................... 2
Opalescent areas, no details of iris visible, size of pupil barely discernible ....................... 3
Opaque, iris invisible ............................................................................................................ 4
Area of cornea affected
One quarter (or less) but no zero ........................................................................................ 1
Greater than one quarter but less than half.......................................................................... 2
Greater than half, but less than three quarters .................................................................... 3
Greater than three quarters, up to whole area ..................................................................... 4

IRIS
Normal .................................................................................................................................... 0
Folds above normal, congestion, swelling, circumcorneal injection (any or all of these
or combination of any thereof) iris still reacting to light (sluggish reaction is positive).......... 1
No reaction to light, haemorrhage, gross destruction (any or all of these) ........................... 2

CONJUNCTIVAE
Redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris)
Vessels normal ....................................................................................................................... 0
Vessels definitely injected above normal ............................................................................... 1
More diffuse, deeper crimson red, individual vessels not easily discernible ......................... 2
Diffuse beefy red .................................................................................................................... 3
Chemosis
No swelling ...............................................................................................................................0
Any swelling above nominal (including nictitating membrane) ................................................. 1
Obvious swelling with partial eversion of lids ......................................................................... 2
Swelling with lids about half closed ..... ................................................................................... 3
Swelling with lids about half closed to completely closed .. .................................................... 4

OCULAR DISCHARGE
No discharge ..............................................................................................................................0
Any amount different from normal (does not include amounts
observed in inner canthus of normal animals) ...........................................................................1
Discharge with moistening of the lids and hairs just adjacent to lids ........... ............................ 2
Discharge with moistening of the lids and hairs, and considerable area around the eye ........ 3

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Results 2

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 h

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

According to the EC-standards (as published in the Official Journal of the European Communities, L 110 A, Volume 36, 4 May 1993) and according to the Globally Harmonised System of Classification and Labelling of Chemicals (GHS; UN/ECE 2003) Branching Enzyme, batch PPY 27209 is not irritating to (human) eyes.

Executive summary:

Branching enzyme, batch PPY 27209 was tested for acute eye irritating properties in an experiment with three albino rabbits, according to EC Directive 92/69/EC, method B.5 and OECD Guideline no. 405. Branching Enzyme, batch PPY 27209 caused slight redness of the conjunctivae in the three rabbits. At 48 h after treatment, all eye effects had cleared completely.

According to the EC-standards (as published in the Official Journal of the European Communities, L 110 A, Volume 36, 4 May 1993) and according to the Globally Harmonised System of Classification and Labelling of Chemicals (GHS; UN/ECE 2003) Branching Enzyme, batch PPY 27209 is not irritating to (human) eyes.