Trehalose

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 Mar. 2001 -19 Jul. 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Trehalose

EC Number:

202-739-6

EC Name:

Trehalose

Cas Number:

99-20-7

Molecular formula:

C12H22O11

IUPAC Name:

trehalose

Test material form:

solid: crystalline

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 9F22
- Expiration date of the lot/batch: 22 Jun 2002
- Purity test date: not stated

RADIOLABELLING INFORMATION (if applicable)
not applicable

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Stability under test conditions: assumed stable
- Solubility and stability of the test substance in the solvent/vehicle: assumed stable and soluble
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: assumed non-reactive

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
Trehalose was administered as supplied by the Sponsor.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Highgate Farm, Market Rasen, Lincolnshire, England
- Age at study initiation: 9 weeks
- Weight at study initiation: 2.6 to 2.9 kg
- Housing: individually
- Diet (e.g. ad libitum): standard laboratory diet, ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: seven days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 +- 2°C
- Humidity (%): 30 - 70 %
- Air changes (per hr): not stated
- Photoperiod (hrs dark / hrs light): 12 / 12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.1 mL of volume applied, correcponding to mean weight of 93 mg

Duration of treatment / exposure:

no fixed exposure time, no washing

Observation period (in vivo):

3 days

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): not done
- Time after start of exposure:

SCORING SYSTEM:

Ocular irritation was assessed using the prescribed numerical system:
Cornea Opacity: degree of density (area most dense taken for reading)
0 No ulceration or opacity
1 Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details
of iris clearly visible
2 Easily discernible translucent areas, details of iris slightly obscured
3 Nacreous areas, no details of iris visible, size of pupil barely discernible
4 Opaque cornea, iris not discernible through the opacity

Area of cornea involved:
0 None
1 One quarter (or less) but not zero
2 Greater than one quarter, but less than half
3 Greater than half. but less than three quarters
4 Greater than three quarters, up to whole area

Iris:
0 Normal
1 Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or
injection, any of these or combination of any thereof, iris still reacting to light (sluggish
reaction is positive)
2 No reaction to light, haemorrhage, gross destruction (any or all of these) 2

Conjunctivae Redness: (refers to the most severe reading of palpebral and bulbar conjunctivae, as
compared to the control eye)
0 Blood vessels normal
1 Some blood vessels definitely hyperaemic (injected)
2 Diffuse, crimson colour, individual vessels not easily discernible ·
3 Diffuse beefy red

Chemosis: (lids and/or nictating membranes)
0 No swelling
1 Any swelling above nonnal (includes nictating membranes)
2 Obvious swelling with partial eversion of lids
3 Swelling with lids about half-closed
4 Swelling with lids more than half-closed

TOOL USED TO ASSESS SCORE: handheld lamp

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Results 5

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

fully reversible

Results 6

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritant / corrosive response data:

A diffuse crimson colouration of the conjunctivae was seen in one animal from approximately one hour
after instillation, resolving completely by two days after instillation. Transient hyperaemia of the blood
vessels of the conjunctivae was observed in the remaining two animals from approximately one hour
after instillation, resolving completely by either one or two days after instillation.

Other effects:

There were no signs of toxicity or ill health in any rabbit during the observation period.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

A single instillation of Trehalose into the eye of the rabbit elicited transient, very slight to well-defined
conjunctival irritation. Other effects related to irritation to the eyes were not observed.

Executive summary:

A study was performed to assess the eye irritation potential of Trehalose to the rabbit. The method

followed was that described in:

EEC Methods for the determination of toxicity, Annex to Directive 92/69/EEC (Official Journal

No. L383A 29.12.92), Part B, Method B.5. Acute toxicity (eye irritation).

OECD Guideline for the Testing of Chemicals No. 405, "Acute Eye Irritation/Corrosion", Adopted

24 February 1987.

EPA Health Effects Test Guidelines OPPTS 870.2400 Acute Eye Irritation EPA 712-C-98-195.

August 1998.

Three rabbits were each administered a single ocular dose of a volume of 0.1 ml of the test substance

(mean weight 93 mg) and observed for three days after instillation.

A single instillation of Trehalose into the eye of the rabbit elicited transient very slight to well-defined

conjunctiva! irritation, resolving completely by two days after treatment.

The means of scores for the ocular reactions at approximately 24, 48 and 72 hours after administration,

calculated separately for each animal. are summarised below:

Animal number	Corneal opacity	Iridial lesions	Conjunctival
			Redness	Chemosis
#1 (2558)	0	0	0.3	0
#2 (2556)	0	0	0.3	0
#3 (2557)	0	0	0	0