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 reports

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
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 the test chemical on eye, when exposed by the ocular route in rabbits.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study report): methyl phenylacetate
- Molecular Formula: C9H10O2
- Molecular Weight: 150.176 g/mol
- Substance type: Organic
- Physical state: Liquid

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Sex: male
- Source:Procured from LIVEON BIOLABS PVT. LTD., Karnataka, India
- Age at study initiation:3.0 to 4.5 Months (Approximately)
- Weight at study initiation: Minimum: 1.568 kg and Maximum: 1.742 kg (Prior to Treatment)
- Health Status :Healthy young adult animals were used for the study.
- Housing:The animals were housed individually in stainless steel cages.
- Room Sanitation:The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
- Cages and water bottle:All the cages and water bottles were changed minimum twice a week.
- Diet (e.g. ad libitum):All animals were provided conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune) ad libitum.
- Water (e.g. ad libitum):Aqua guard filtered tap water was provided ad libitum.
- Acclimation period: Rabbits were acclimatised to the test conditions for a period of 6 days (Animal No.-1) and 9 days (Animal No. 2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 19.00 °C and Maximum: 20.80 °C
- Humidity (%):Minimum: 45.10 % and Maximum: 65.80 %
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12:12

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):N/A

VEHICLE
- Amount(s) applied (volume or weight with unit):N/A
- Concentration (if solution):N/A
- Lot/batch no. (if required):N/A
- Purity:N/A
Duration of treatment / exposure:
24 hrs
Observation period (in vivo):
All the animals were observed at 1, 24, 48 and 72 hours after instillation of test item.
Duration of post- treatment incubation (in vitro):
no data available
Number of animals or in vitro replicates:
3 male 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 hours

SCORING SYSTEM:Draize Method

TOOL USED TO ASSESS SCORE: Ophthalmoscope and fluorescent strips.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal: #1,#2 and #3
Time point:
other: After 1, 24, 48 and 72 Hours
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: After 1, 24, 48 and 72 Hours
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: After 1, 24, 48 and 72 Hours
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: After 1, 24, 48 and 72 Hours
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 1, 24, 48 and 72 Hours
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
iris score
Basis:
animal: #1,#2 and #3
Time point:
other: After 1, 24, 48 and 72 Hours
Score:
0
Max. score:
2
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: After 1 and 24 Hours
Score:
1
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
other: After 1 and 24 Hours
Score:
1
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: After 1 and 24 Hours
Score:
1
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
conjunctivae score
Basis:
animal: #1,#2 and #3
Time point:
other: After 48 and 72 Hour
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: After 1 Hours
Score:
1
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: After 1 Hours
Score:
1
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: After 1 Hours
Score:
1
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
chemosis score
Basis:
animal: #1,#2 and #3
Time point:
other: After 1, 24, 48 and 72 Hours
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritant / corrosive response data:
Untreated eye of all the three rabbits was normal throughout the experimental period of 72 hours.

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 the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all the animals.

Observation at 24 hours after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: No swelling (normal) was observed in all the animals.
At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 0% damage in all the animals.

Observation at 48 and 72 hours after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Blood vessels normal in all the animals; Chemosis: No swelling (normal) was observed in all the animals.

The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours (at treated site) for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively.
Other effects:
Clinical Observation
No systemic toxicity was observed in treated rabbits during the experimental period

Mortality
No mortality was observed during the observation period.

Body weight
Increase in body weights in all the treated animals at termination when compared to day 0 was observed

Any other information on results incl. tables

Table 1 : Individual Animal Eye Irritation Scores

 

In Treated area Dose:0.1 ml of as such test item                                                 Sex:Male

Animal numbers

1

2

3

Application site

Right

Right

Right

Eye reactions

At hours

At hours

At hours

*

1

24

48

72

*

1

24

48

72

*

1

24

48

72

Corneal opacity

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

Iris

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctiva

0

1

1

0

0

0

1

1

0

0

0

1

1

0

0

Chemosis

0

1

0

0

0

0

1

0

0

0

0

1

0

0

0

Corneal damage %

0

./.

0

0

./.

0

0

./.

0

In Treated area Dose:Untreated                                                                             Sex:Male

Animal Numbers

1

2

3

Application Side

Left

Left

Left

Eye Reactions

At hour

At hour

At hour

*

1

24

48

72

*

1

 24

48

72

*

1

24

48

72

Corneal Opacity

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

Iris

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

Chemosis

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Corneal Damage%

0

./.

0

0

./.

0

0

./.

0

Key:*= Pre-exposure eye examination.

      ./.= Not Applicable

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

0.33

0.33

0.33

Chemosis

0.00

0.00

0.00

 

 

Eye Irritation Scores - Mean Values after 24, 48, 72 Hours (Untreated eye)

 

            Animal No.

 Eye Reaction

1

2

3

Corneal Opacity

0.00

0.00

0.00

Iris

0.00

0.00

0.00

Conjunctiva

0.00

0.00

0.00

Chemosis

0.00

0.00

0.00

 

Formula :

 

Mean Eye Irritation Score =

Sum of the Individual Animal Score for eye reaction at 24, 48 and 72 hours

Number of the Observations (3)

Table 2 : Individual AnimalClinicalSigns

 

Sex:Male

Animal No.

Days (Post application observation)

0

1

2

3

1

1

1

1

1

2

1

1

1

1

3

1

1

1

1

Key:1 = Normal

Table 3: Individual Animal Body Weight

Sex :Male

Animal No.

Animal Body Weight (kg)

Prior to application

At termination

1

1.568

1.608

2

1.590

1.674

3

1.742

1.818

Key:kg = Kilogram

Applicant's summary and conclusion

Interpretation of results:
other: not irritating
Conclusions:
The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours (at treated site)for corneal opacity, iris, conjunctiva and chemosis were determined to be 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively. Hence, it was concluded that the test chemical was Non-Irritating to the eyes of Male New Zealand White rabbits under the experimental conditions tested and is Classified as “Category- Not Classified" as per CLP criteria for Classification.
Executive summary:

An Acute Eye Irritation/Corrosion Study for the test substance was performed in Rabbit as per OECD guideline No. 405. 3 male New Zealand White rabbits were used for the study.

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 as such test itemwas instilled in the other (treated) eye of each rabbit. The eye was observed 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 as such test itemwas instilled 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 hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3);0.1 ml of as such test itemwas 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 was normal throughout the experimental period of 72 hours. 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 the animals; Area of Opacity-Zero in all the animals;Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all the animals.

Observation at 24 hours after instillation of test item revealed:Cornea- No ulceration or opacity in all the animals; Area of Opacity-Zero in all the animals;Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis:No swelling (normal) was observed in all the animals.

At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 0% damage in all the animals.Observation at 48 and 72 hours after instillation of test item revealed:Cornea- No ulceration or opacity in all the animals; Area of Opacity-Zero in all the animals;Iris: Normal in all the animals. Conjunctivae - Blood vessels normal in all the animals; Chemosis:No swelling (normal) was observed in all the animals. The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours (at treated site)for corneal opacity, iris, conjunctiva and chemosis were determined to be 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively.

Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed within 72 hours. 

Hence, it was concluded that the test chemical was Non-Irritating to the eyes of Male New Zealand White rabbits under the experimental conditions tested and is Classified as “Category- Not Classified" as per CLP criteria for Classification.