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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The study was conducted between 05 June 2015 and 16 June 2015.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study is considered to be a reliability 1 as it has been conducted according to OECD Test Guideline 405 using an in vivo method and in compliance with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Liquid
Details on test material:
Identification: FRET 08-0338
Chemical name: 4,7-Methano-1H-inden-5-ol, octahydro-3,4,5-trimethyl and 4,7-Methano-1H-inden-5-ol, octahydro-2,4,5-trimethyl and isomers
CAS number: 1340502-93-3 & 1340502-69-3
Intended use: Industrial fragrance
Description: Clear liquid
Storage conditions: Room temperature in the dark

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
The animal for this study was selected from a stock supply of healthy adult rabbits of the New Zealand White strain. The animal weighed 2.71 kg and was approximately 23 weeks of age, prior to treatment (Day 1). She had been acclimatised to the experimental environment for a period of approximately ten weeks prior to the start of the study.
The animal was housed individually in a plastic cage with perforated floors and was offered 150 g of a standard laboratory rabbit diet per day; drinking water was provided ad libitum.
The batch of diet used for the study is analysed for nutrients, possible contaminants and micro-organisms likely to be present in the diet and which, if in excess of specified amounts, might have an undesirable effect on the test system. A dietary supplement of hay was offered during acclimatisation until three days prior to dose instillation, for the remainder of acclimatisation and throughout the study observation period wholemeal bread was offered.
During the acclimatisation and study period the animal was given a small, soft white untreated wood block for environmental enrichment, replaced at appropriate intervals.
Results of routine physical and chemical examination of drinking water, as conducted by the supplier are made available to Huntingdon Life Sciences.
Animal room environmental controls were set to maintain temperature within the range 16 to 20°C, and relative humidity within 40 to 70%. These environmental parameters were recorded and the permanent record archived with other departmental raw data. Lighting was controlled by means of a time switch to give 12 hours of artificial light (06:00 to 18:00 GMT) in each 24 hour period.
The animal was identified by a numbered tag placed through the edge of one ear. This identification was unique within the Department throughout the duration of the study. The cage was identified by a coloured label displaying the study number and animal number.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
The test substance was not removed
Observation period (in vivo):
Seven days after instillation
Number of animals or in vitro replicates:
One
Details on study design:
Treatment procedure
The eyes of the animal were examined prior to instillation of the test substance to ensure that there was no pre-existing corneal damage, iridial inflammation or conjunctival irritation. The animal was gently restrained and the dose was instilled into the right eye by pulling the lower eyelid away from the eye ball to form a cup into which the test substance was dropped. The eyelids were then gently held together for one second before releasing; the left eye remained untreated.
A single animal was treated in advance; the presence of a severe effect in this animal prevented further animals being committed to the study.

Clinical signs
The behaviour of the rabbit was observed immediately following instillation of the test substance to allow assessment of the initial pain response.
The animal was returned to the cage and checked at least twice during the first hour after dosing and at regular intervals throughout the day to ensure no severe injury passed unnoticed. Ocular reactions to treatment were assessed 1, 24, 48 and 72 hours and seven days after treatment, according to the criteria below. Reactions not included overleaf were described in detail.

Ocular responses
The untreated eye was used as a comparison with the treated eye during assessment of ocular lesions.
A pencil beam torch was available for use to facilitate inspection of the eyes.

Termination
Following completion of the observation period the animal was humanely killed by an intravenous injection of sodium pentobarbital.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: 24, 48 and 72 hours
Score:
1
Reversibility:
not reversible
Remarks on result:
other: Pannus formation (corneal neovascularisation) is considered to be an irreversible effect
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: 24, 48 and 72 hours
Score:
0.3
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #1
Time point:
other: 24, 48 and 72 hours
Score:
2
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 24, 48 and 72 hours
Score:
0.3
Irritant / corrosive response data:
Ocular responses
A crimson-red conjunctival appearance was evident throughout the study period and very-slight or slight chemosis was apparent during the first 24 hours after instillation, with discharge evident at 1 hour after instillation only. Iritis was evident during the first 24 hours after instillation and also seven days later.
Scattered or diffuse areas of opacity covering up to the entire corneal surface were apparent throughout the first 72 hours after instillation. Seven days after instillation an easily discernible translucent area of opacity covering approximately one quarter of the corneal surface and an area of scattered or diffuse opacity covering approximately half the corneal surface were evident. In addition, two areas of pannus formation (corneal neovascularisation) were apparent; pannus formation is considered to be an irreversible effect and the animal was humanely killed immediately after this observation.
Instillation of the test material gave rise to no initial pain response.
Other effects:
Clinical signs
There was no sign of toxicity or ill health in the study rabbit during the observation period.

Any other information on results incl. tables

Mean values for ocular lesions for Kay and Calandra classification

Mean irritation scores after instillation of FRET 08-0338

Area of eye

1 hour

24 hours

48 hours

72 hours

7 days

Cornea

20

20

20

15

20

Iritis

5

5

0

0

5

Conjunctiva

12

6

4

4

4

Total mean score

37

31

24

19

29

 

Mean values for ocular lesions for EC (Regulation 1272/2008) and GHS classification

24, 48 and 72 hours after instillation of FRET 08-0338

Animal number and sex

Corneal opacity

Iridial lesions

Redness of conjunctiva

Chemosis

188 Female

1.0

0.3

2.0

0.3

 

Grades for ocular irritation responses following instillation of FRET 08-0338

Animal number and sex: 188 F

Pain evaluation response: 0

Region of the eye

Response

Grade of response at time after instillation

Hours

Day

1

24

48

72

7

Cornea

Opacity (A)

1

1F+

1F+

1F+

2F+AP

Area (B)

4

4

6

3

1

Ulceration

-

-

-

-

-

Stippling

-

-

-

-

-

Corneal score (A x B x 5)

20

20

20

15

10 + 10

Iris

Value (C)

1

1

0

0

1

Iridial score(C x 5)

5

5

0

0

5

Conjunctiva

Redness (D)

2

2

2

2

2

Chemosis (E)

2

1

0

0

0

Discharge (F)

2

0

0

0

0

Necrosis

-

-

-

-

-

Ulceration

-

-

-

-

-

Conjunctival score ((D + E + F) x 2)

12

6

4

4

4

F = Female

F+ = Fluorescein positive

A = Additional area of opacity Grade 1 and Area 2 (fluorescein negative)

P = Two areas of pannus formation

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye)
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The highest total mean score was 37 occurring at the 1 hour observation; accordingly under the criteria Kay and Calandra (1962) FRET 08-0338 was classified as “moderately irritating” to the eye.
Pannus formation (corneal neovascularisation) is considered to be an irreversible effect; accordingly, FRET 08-0338 was classified as Category 1 in accordance with European Commission regulation 1272/2008.
Executive summary:

The eye irritation potential of the test substance (FRET 08-0338) was assessed according to OECD Test Guideline 405 using an in vivo method. Pannus formation (corneal neovascularisation) is considered to be an irreversible effect; accordingly, FRET 08-0338 was classified as Category 1 in accordance with European Commission regulation 1272/2008.