Registration Dossier

Toxicological information

Eye irritation

Currently viewing:

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from experimental study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report date:
2014

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Principles of method if other than guideline:
The objective of the study was to assess the irritant and/or corrosive effects of test chemical on eye when exposed by an ocular route in rabbits.
GLP compliance:
not specified

Test material

Constituent 1
Chemical structure
Reference substance name:
3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexan-1-ol
EC Number:
222-294-1
EC Name:
3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexan-1-ol
Cas Number:
3407-42-9
Molecular formula:
C16H28O
IUPAC Name:
.
Details on test material:
- Name of test material (as cited in study report):3-(5, 5, 6-trimethylbicyclo [2.2.1] hept-2-yl) cyclohexan-1-ol - Molecular formula : C16H28O- Molecular weight :236.39 g/mol- Substance type:Organic- Physical state:Colourless Liquid- Lot/batch No.:Lot 1/14- Storage condition of test material:Store in tightly closed container in a dry, cool and well-ventilated place. Protect from light, including direct sunrays. - Other:Handling and DisposalSafety precautions : Avoided contact with skin and eyes. Kept away from sources of ignition. Use aprons, masks, caps, gloves and goggles to ensure the health and safety of the personnel.Disposal : The remaining unused test item was disposed as per internal SOPs of sa-FORD.

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS- Source:Procured from RABBI ROOF, Hyderabad
.- Age at study initiation:4 to 5 Months (Approximately)
- Weight at study initiation: 1.999-2.268 kg
- Housing:The animals were housed individually in stainless steel cages in a controlled environment.
- Diet (e.g. ad libitum): Conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune), ad libitum
- Water (e.g. ad libitum):Aqua guard filtered tap water, ad libitum-
Acclimatization period: Rabbits were acclimatized to the test conditions for a period of 5 days (Animal No. 1) and 9 days (Animal No. 2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.20-22.60°C
- Humidity (%): 42.00-74.70%
- Air changes (per hr):More than 12 changes per hour
- Photoperiod (hrs dark / hrs light):12-hrs light/12-hrs dark

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml
- Concentration (if solution): No data available

VEHICLE
- Amount(s) applied (volume or weight with unit): No data available
- Concentration (if solution): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
All the animals were observed at 1, 24, 48, 72 hours and on day 7 after instillation of test item.
Number of animals or in vitro replicates:
3 female rabbits
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated eye of rabbit was washed with normal saline
- Time after start of exposure: 24 hrs

SCORING SYSTEM: Grading of irritation lesions was carried out as per Draize Method

TOOL USED TO ASSESS SCORE: Ophthalmoscope and fluorescein strips.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal: #1,#2 and #3
Time point:
other: 24,48,72 hrs and 7 days
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
Untreated (Control Eye)
Irritation parameter:
iris score
Basis:
animal: #1,#2 and #3
Time point:
other: 24,48,72 hrs and 7 days
Score:
0
Max. score:
2
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
Untreated (Control Eye)
Irritation parameter:
conjunctivae score
Basis:
animal: #1,#2 and #3
Time point:
other: 24,48,72 hrs and 7 days
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
Untreated (Control Eye)
Irritation parameter:
chemosis score
Basis:
animal: #1,#2 and #3
Time point:
other: 24,48,72 hrs and 7 days
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
Untreated (Control Eye)
Irritation parameter:
cornea opacity score
Basis:
animal: #1,#2 and #3
Time point:
other: After 24, 48, 72 Hours
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eye
Irritation parameter:
iris score
Basis:
animal: #1,#2 and #3
Time point:
other: After 24, 48, 72 Hours
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eye
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: After 24 Hours
Score:
1.67
Max. score:
4
Reversibility:
not specified
Remarks on result:
other: For treated eyes
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
other: After 48 Hours
Score:
1
Max. score:
4
Reversibility:
not specified
Remarks on result:
other: For treated eyes
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: After 72 Hours
Score:
4
Max. score:
3
Reversibility:
not specified
Remarks on result:
other: For treated eyes
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: After 24 Hours
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
other: For treated eye
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: After 48 Hours
Score:
1
Max. score:
3
Reversibility:
not specified
Remarks on result:
other: For treated eye
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: After 72 Hours
Score:
1
Max. score:
3
Reversibility:
not specified
Remarks on result:
other: For treated eyes
Irritant / corrosive response data:
The following were observed in treated rabbits.In the initial test, 0.1 ml of test item was applied into the conjunctival sac of the right eye of animal no.1 whereas the left eye of the rabbit served as the control. As animal no. 1 showed no severe ocular lesions a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 0.1 ml of test item was instilled into the conjunctival sac of right eye of both the rabbits and left eye served as the control. Ocular lesions were seen in animal no. 2 and 3 at till 72 hour observation which recovered at day 7.Untreated eye of all the three rabbits was normal throughout the experimental period of 7 days.The following grading scores were observed in treated eye of tested rabbits.Observation at 1 hour after instillation of test item revealed:Cornea: No ulceration or opacity in all 3 animals; Area of Opacity: Zero in all 3 animals; Iris: Normal in all 3 animals. Conjunctivae: Blood vessels normal was seen in all 3 animals; Chemosis: No swelling (Normal) was seen in animal no. 1 whereas some swelling above normal (includes nictitating membranes) was seen in animal no. 2 and 3.Observation at 24 hours after instillation of test item revealed: Cornea: No ulceration or opacity in all 3 animals; Area of Opacity: Zero in all 3 animals; Iris: Normal in all 3 animals; Conjunctivae: Some blood vessels definitely hyperaemic (injected) was seen in animal no. 1 and 2 whereas obvious swelling with partial eversion of lids was seen in animal no. 3; Chemosis: No swelling (Normal) was seen in animal no. 1 whereas some swelling above normal (includes nictitating membranes) was seen in animal no. 2 and 3.At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 0 %, 15 % and 20 % damage in animal no. 1, 2 and 3 resectively. Observation at 48 and 72 hours after instillation of test item revealed: Cornea: No ulceration or opacity in all 3 animals; Area of Opacity: Zero in all 3 animals; Iris: Normal in all 3 animals; Conjunctivae: Some blood vessels definitely hyperaemic (injected) was seen in animal no. 2 whereas obvious swelling with partial eversion of lids was seen in animal no. 1 and 3; Chemosis: No swelling (Normal) was seen in animal no. 1 whereas some swelling above normal (includes nictitating membranes) was seen in animal no. 2 and 3.Observation at day 7 after instillation of test item revealed: Cornea: No ulceration or opacity in all 3 animals; Area of Opacity: No opacity was seen in all 3 animals; Iris: Normal in all 3 animals. Conjunctivae: Blood vessels normal was seen in all 3 animals; Chemosis: No swelling (Normal) was seen in all 3 animals.The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 1.67, 0.00; 0.00, 0.00, 1.00, 1.00 and 0.00, 0.00, 2.00, 1.00, respectively.
Other effects:
No systemic toxicity or mortality was observed in treated rabbits during the experimental period. All the rabbits were observed with a body weight gain on day 7 as compared to day 0.

Any other information on results incl. tables

Table 1 : Individual Animal Eye Irritation Scores

 

In Treated area Dose:0.1 ml of test item  Sex:Female

Animal Numbers

1

2

3

Application Side

Right

Right

Right

Eye Reactions

At hour

At hour

At hour

*

1

24

48

72

7 days

*

1

24

48

72

7 days

*

1

24

48

72

7 days

Corneal Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Area of Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Iris

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctiva

0

1

1

2

2

0

0

1

1

1

1

0

0

1

2

2

2

0

Chemosis

0

0

0

0

0

0

0

1

1

1

1

0

0

1

1

1

1

0

Corneal Damage%

0

15

20

 

 

Dose:Untreated (Control Eye)                                                                         Sex:Female

Animal Numbers

1

2

3

Application Side

Left

Left

Left

Eye Reactions

At hour

At hour

At hour

*

1

24

48

72

Day 7

*

1

24

48

72

Day 7

*

1

24

48

72

Day 7

Corneal Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Area of Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Iris

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctiva

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Chemosis

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Corneal Damage%

0

0

0

Key:*= Pre-exposure eye examination.

 

 

 

Table 1 (Continued):Eye Irritation Scores - Mean Values after 24, 48, 72 Hours (Treated eye)

            Animal No.

 Eye Reaction

1

2

3

Corneal Opacity

0.00

0.00

0.00

Iris

0.00

0.00

0.00

Conjunctiva

1.67

1.00

2.00

Chemosis

0.00

1.00

1.00

 

 

Formula :

 

Mean Eye Irritation Score =

                          

Sum of the Individual Animal Score for eye reactionat24, 48 and 72 hours

Number of the Observations (3)

 

Table 2: Individual AnimalClinicalSigns

 

Sex:Female

Animal No.

Days (Post application observation)

0

1

2

3

4

5

6

7

1

1

1

1

1

1

1

1

1

2

1

1

1

1

1

1

1

1

3

1

1

1

1

1

1

1

1

Key:1 = Normal

 

  

Table 3: Individual Animal Body Weight

Sex :Female

Animal No.

Animal Body Weight (kg)

Prior to application

At termination

1

2.268

2.361

2

1.999

2.201

3

2.092

2.213

Key:kg = Kilogram

Applicant's summary and conclusion

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
Under the experimental conditions tested, was considered as mildly irritating to eyes to New Zealand White female rabbit eyes.
Executive summary:

Acute eye irritation/corrosion study was conducted in rabbits to evaluate the eye irritant nature of the test chemical. The study was performed as per OECD 405 Guidelines using 3 female New Zealand White rabbits.

Rabbits free from injury of eye were selected for the study. The eyes of all the rabbits were examined 24 hours prior to treatment. One eye of each rabbit served as control and other as treated. Control eye was left untreated whereas; 0.1 ml of test item was instilled in the other (treated) eye of each rabbit. The eye was observed at 1, 24, 48, 72 hour, day 7, day 14 and day 21 for animal no. 1 and 3 whereas at 1, 24, 48 and 72 hour after test item instillation. Ophthalmoscope was used for scoring of eye lesions.

In the initial test, 0.1 ml of test item (as such) was applied into the conjunctival sac of the right eye of animal no.1 whereas the left eye of the rabbit served as the control. As animal no. 1 showed no severe ocular lesions a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 0.1g of test item (in pulverized form) was instilled into the conjunctival sac of right eye of both the rabbits and left eye served as the control.

Untreated eye of all the three rabbits were normal throughout the experimental period of 7 days.

The following grading scores were observed in treated eye of tested rabbits.

Observation at 1 hour after instillation of test item revealed: Cornea: No ulceration or opacity in all 3 animals; Area of Opacity: Zero in all 3 animals; Iris: Normal in all 3 animals. Conjunctivae: Blood vessels normal was seen in all 3 animals; Chemosis: No swelling (Normal) was seen in animal no. 1 whereas some swelling above normal (includes nictitating membranes) was seen in animal no. 2 and 3.

Observation at 24 hours after instillation of test item revealed: Cornea: No ulceration or opacity in all 3 animals; Area of Opacity: Zero in all 3 animals; Iris: Normal in all 3 animals; Conjunctivae: Some blood vessels definitely hyperaemic (injected) was seen in animal no. 1 and 2 whereas obvious swelling with partial eversion of lids was seen in animal no. 3; Chemosis: No swelling (Normal) was seen in animal no. 1 whereas some swelling above normal (includes nictitating membranes) was seen in animal no. 2 and 3.At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 0 %, 15 % and 20 % damage in animal no. 1, 2 and 3 respectively.

Observation at 48 and 72 hours after instillation of test item revealed: Cornea: No ulceration or opacity in all 3 animals; Area of Opacity: Zero in all 3 animals; Iris: Normal in all 3 animals; Conjunctivae: Some blood vessels definitely hyperaemic (injected) was seen in animal no. 2 whereas obvious swelling with partial eversion of lids was seen in animal no. 1 and 3; Chemosis: No swelling (Normal) was seen in animal no. 1 whereas some swelling above normal (includes nictitating membranes) was seen in animal no. 2 and 3.

Observation at day 7 after instillation of test item revealed: Cornea: No ulceration or opacity in all 3 animals; Area of Opacity: No opacity was seen in all 3 animals; Iris: Normal in all 3 animals. Conjunctivae: Blood vessels normal was seen in all 3 animals; Chemosis: No swelling (Normal) was seen in all 3 animals. The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 1.67, 0.00; 0.00, 0.00, 1.00, 1.00 and 0.00, 0.00, 2.00, 1.00, respectively.

Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of all 3 animals was observed at day 7. Hence, the test chemical is considered as mildly irritating to the eyes of female New Zealand White rabbits, and thus the substance is classified as an eye irritant in Category 2 as per the CLP regulation.