Registration Dossier

Administrative data

Description of key information

Skin Irritation:

Acute Dermal Irritation/corrosion Study of test chemical in Rabbits, was performed as per OECD guideline No. 404. A volume of 0.5 ml of test item (as such) was applied to the skin for 4 hours and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hours post patch removal, evaluated and graded as per draize method. The individual mean score at 24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively Hence, it was concluded that the test chemical was Non-Irritating to the skin of female New Zealand White rabbits under the experimental conditions tested. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

Eye Irritation:

Acute Eye Irritation/Corrosion Study of Test chemical in Rabbits, was performed as per OECD guideline No. 405. The. 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. 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, 0.67, 0.00; 0.00, 0.00, 0.67, 0.00 and 0.00, 0.00, 0.67, 0.00, respectively Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed at 72 hours. Hence, the test chemical was considered as Non-Irritating to the eyes of male New Zealand White rabbits under the experimental conditions tested. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin 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 report
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal 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 after dermal application on the intact skin of rabbits.
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
Species :Rabbits (Oryctolagus cuniculus)
Strain :New Zealand White
Age :4 to 4.5 Months (Approximately)
Sex :Female
Number of Animals:Three
Supplier/Source:Procured from LIVEON BIOLABS PVT. LTD. (CPCSEA Reg. No. 1610/RO/bc12/CPCSEA).
Health Status :Healthy young adults rabbits were used for the study. Females were nulliparous and non pregnant.
Body weight of animals: Minimum: 2.168 kg and Maximum: 2.404 kg (Prior to Treatment)

Acclimatisation: Rabbits were acclimatised to the test conditions for a period of 6 days (Animal No.-1) and 8 days (Animal No.-2 and 3) prior to the application of the test item.
Identification: During acclimatization marking was done with non toxic marker pen in the inside of left ear of rabbits and after acclimatization, animals were marked with permanent number in the inner side of right ear of rabbits. Permanent marker and cage card was used for identification. The individual cage cards were labelled with at least project No., species, strain, group, sex, animal number, experiment start and end date.

Husbandry Conditions
Diet : All animals were provided conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No.: 200004.
Water : Aqua guard filtered tap water was provided ad libitum.
Husbandry : 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.

Experimental Room Condition
Temperature : Minimum: 20.50 °C, Maximum: 22.50 °C
Relative humidity: Minimum: 51.50 %, Maximum: 66.30 %
Light-dark-rhythm: 12:12
Air Changes: More than 12 changes per hour
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):0.5 ml
- Concentration (if solution): No data available

VEHICLE
- Amount(s) applied (volume or weight with unit):0.5 ml distilled water
- Concentration (if solution): No data available
- Lot/batch no. (if required): No data available- Purity: No data available
Duration of treatment / exposure:
4 hours
Observation period:
72 hours
Number of animals:
3 female rabbits
Details on study design:
TEST SITE
- Area of exposure: approximately 6 X 6 cm at contralateral sites
- Type of wrap if used: porous gauze dressing and non-irritating tape (Micropore 3”)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): residual test item was removed by using cotton soaked in distilled water
- Time after start of exposure: 4hr

Control site: 0.5 ml distilled water was applied at control site.
Irritation parameter:
erythema score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: Not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: Not applicable
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method. The individual mean score at 24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively.

No erythema and edema (skin irritation) were observed at the end of 72 hour observation period after patch removal.
Other effects:
Clinical Observation
No systemic toxicity was observed at treated rabbits during the experimental period.
Mortality
No mortality was observed during the observation period.

Table 1

Skin Reaction

 

In Treated area Dose:0.5 ml of test item                                                                      Sex:Female

 

Animal

No.

Test

Treated

 area*

Erythema score

Oedema score

1h

24h

48h

72h

1h

24h

48h

72h

1

Initial

Right

0

0

0

0

0

0

0

0

2

Confirmatory

Left

0

0

0

0

0

0

0

0

3

Left

0

0

0

0

0

0

0

0

 

 

 

 

 

In Control areaDose:0.5 ml of distilled water                                                         Sex:Female

 

Animal

No.

Test

Treated area*

Erythema score

Oedema score

1h

24h

48h

72h

1h

24h

48h

72h

1

Initial

Left

0

0

0

0

0

0

0

0

2

Confirmatory

Right

0

0

0

0

0

0

0

0

3

Right

0

0

0

0

0

0

0

0

Key:h = Hour.

 

Erythema                                                                                                       Oedema

0 =No erythema                                                                                           0 =No oedema


Table 1 Continued…

Mean Individual Animal Score at 24, 48 and 72 hours

 

                     Animal Number                  

Observations                      

1

2

3

Erythema

0.00

0.00

0.00

Oedema

0.00

0.00

0.00

 

Table 2

Individual Animal BodyWeight

Sex:Female

Animal

No.

Body Weight (kg)

Prior to Dosing

At termination

1

2.292

2.340

2

2.404

2.464

3

2.168

2.248

Table 3

Individual AnimalClinical Signs

Sex:Female

Animal

No.

Days (Post dosing Observation)

0

1

2

3

1

1

1

1

1

2

1

1

1

1

3

1

1

1

1

Key: ./. = Not Applicable. 1 = Normal.


Interpretation of results:
other: not irritating
Conclusions:
The individual mean score at 24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively.Hence, the test chemical was regarded as non-Irritating to the skin of male New Zealand White rabbits under the experimental conditions tested and is thus not classified as a skin irritant.
Executive summary:

Acute Dermal Irritation/corrosion Study of test chemical was performed as per OECD guideline No. 404 using three healthy young adult female.

Rabbits with good intact skin were selected for the study. The hairs of all the rabbits were clipped at contralateral sites, approximately 24 hours prior to treatment. A dose of 0.5 ml of test item (as such) was applied to the skin, over an area of approximately 6 x 6 cm clipped of hair on one side of rabbits. The other untreated side was kept as control area and 0.5 ml of distilled water was applied at this site. At the end of 4 hours, the gauze patch was removed and test item application site was wiped with water without altering the integrity of the epidermis. 

Initially, the test item was applied to the clipped area of skin of one rabbit. The test site was covered with gauze patch. After 4 hours of exposure, animal no. 1 revealed no erythema and oedema at 1, 24, 48 and 72 hours observation. Hence the confirmatory test was conducted on additional two animals (no. 2 and 3) to confirm the non-irritant nature of the test item.

After 4 hours of exposure, animal no. 2 and 3 revealed no erythema and oedema at 1, 24, 48 and 72 hours observation.

The patch was removed after 4 hours and animals were observed for erythema and oedema at 1, 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method.

The individual mean score at 24, 48 and 72 hours for animal nos. 1, 2 and 3 were 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively.  

Hence, it was concluded that the test chemical was Non-Irritating to the skin of female New Zealand White rabbits under the experimental conditions tested and Classified as “Category- Not Classified as Skin Irritant” as per CLP Classification.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
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
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 test chemical on eye when exposed by an ocular route in rabbits.
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Species : Rabbit (Oryctolagus cuniculus)
Strain : New Zealand White
Age : 2.5 to 4.0 Months (Approximately)
Sex : Male
Number of Animals: Three
Supplier/Source: Procured from Liveon Biolabs Pvt. Ltd., Karnataka, India (CPCSEA Reg. No. 1610/RO/bc12/CPCSEA).
Health Status : Healthy young adult animals were used
Body weight of animals : Minimum: 2.026 kg and Maximum: 2.308 kg (Prior to Treatment)

Acclimatisation: Rabbits were acclimatised to the test conditions for a period of 7 days (Animal No.-1) and 10 days (Animal No. 2 and 3) prior to the application of the test item.
Identification : During acclimatization marking was done with non toxic marker pen in the inside of left ear of rabbits and after acclimatization, animals were marked with permanent number in the inner side of right ear of rabbits. Permanent marker and cage card was used for identification. The individual cage card was labelled with at least study no., study type, test system, sex, dose, experiment start date and experiment completion date..

Husbandry Conditions
Diet: All animals were provided conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No.: 200004 and 200005.
Water : Aqua guard filtered tap water was provided ad libitum.
Husbandry: 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.

Experimental Room Condition
Temperature : Minimum: 20.10 °C, Maximum: 22.20 °C
Relative humidity: Minimum: 57.20 % Maximum: 69.20%
Light-dark-rhythm: 12:12
Air Changes: More than 12 changes per hour
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.1 ml test item (as such) was placed in the conjunctival sac of three rabbits
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
at 1, 24, 48 and 72 hours after instillation of test item.
Number of animals or in vitro replicates:
three 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 hrs

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

TOOL USED TO ASSESS SCORE: Ophthalmoscope and fluorescein strips.
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: Not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal: #1, #2 and #3
Time point:
24/48/72 h
Score:
0.67
Max. score:
3
Reversibility:
fully reversible within: 72 hrs
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: Not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: Not applicable
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
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 seen in all the animals.

Observation at 24 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:No swelling (normal) was seen 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 animal no. 1, 2 and 3.

 
Observation at 48 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:No swelling (normal) was seen in all the animals.

Observation at 72 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 - Blood vessels normal was observed in all the animals;Chemosis:No swelling (normal) was seen in all the animals.

The individual mean score for animal nos. 1, 2 and 3at 24, 48, 72 hoursfor corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.67, 0.00; 0.00, 0.00, 0.67, 0.00 and 0.00, 0.00, 0.67, 0.00, respectively.

Table 1 : Individual Animal Eye Irritation Scores

 

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

Animal Numbers

1

2

3

Application Side

Right

Right

Right

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

1

1

1

0

0

1

1

1

0

0

1

1

1

0

Chemosis

0

1

0

0

0

0

1

0

0

0

0

1

0

0

0

Corneal Damage%

0%

0%

0%

 

 

 

Dose:Untreated (Control Eye)                                                                       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%

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

0.67

0.67

0.67

Chemosis

0.00

0.00

0.00

 

 

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: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

2.308

2.366

2

2.268

2.462

3

2.026

2.126

Key:kg = Kilogram

Interpretation of results:
other: not irritating
Conclusions:
Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed at 72 hours. Hence, the test chemical was considered as “Non Irritant” to New Zealand White male rabbit eyes and being classified as “Not Classified as an Eye Irritant” as per the criteria of CLP regulation.
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 three male 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 and 72 hours after test item instillation. Ophthalmoscope was used for scoring of eye lesions.

 

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 till 24 hour observation hence 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 which were recovered till 72 hours observation.

 

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 seen in all the animals.

Observation at 24 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: No swelling (normal) was seen 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 animal no. 1, 2 and 3.

 

Observation at 48 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: No swelling (normal) was seen in all the animals.

 

Observation at 72 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 - Blood vessels normal was observed in all the animals; Chemosis: No swelling (normal) was seen in all the 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, 0.67, 0.00; 0.00, 0.00, 0.67, 0.00 and 0.00, 0.00, 0.67, 0.00, respectively.

 

Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed at 72 hours. Hence, the test chemical was considered as “Non Irritant” to New Zealand White male rabbit eyes and being classified as “Not Classified as an Eye Irritant” as per the criteria of CLP regulation.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Irritation:

Acute Dermal Irritation/corrosion Study of test chemical was performed as per OECD guideline No. 404 using three healthy young adult female.

Rabbits with good intact skin were selected for the study. The hairs of all the rabbits were clipped at contralateral sites, approximately 24 hours prior to treatment. A dose of 0.5 ml of test item (as such) was applied to the skin, over an area of approximately 6 x 6 cm clipped of hair on one side of rabbits. The other untreated side was kept as control area and 0.5 ml of distilled water was applied at this site. At the end of 4 hours, the gauze patch was removed and test item application site was wiped with water without altering the integrity of the epidermis. 

Initially, the test item was applied to the clipped area of skin of one rabbit. The test site was covered with gauze patch. After 4 hours of exposure, animal no. 1 revealed no erythema and oedema at 1, 24, 48 and 72 hours observation. Hence the confirmatory test was conducted on additional two animals (no. 2 and 3) to confirm the non-irritant nature of the test item.

After 4 hours of exposure, animal no. 2 and 3 revealed no erythema and oedema at 1, 24, 48 and 72 hours observation.

The patch was removed after 4 hours and animals were observed for erythema and oedema at 1, 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method.

The individual mean score at 24, 48 and 72 hours for animal nos. 1, 2 and 3 were 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively.  

Hence, it was concluded that the test chemical was Non-Irritating to the skin of female New Zealand White rabbits under the experimental conditions tested and Classified as “Category- Not Classified as Skin Irritant” as per CLP Classification.

 

Eye Irritation:

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 three male 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 and 72 hours after test item instillation. Ophthalmoscope was used for scoring of eye lesions.

 

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 till 24 hour observation hence 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 which were recovered till 72 hours observation.

 

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 seen in all the animals.

Observation at 24 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: No swelling (normal) was seen 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 animal no. 1, 2 and 3.

 

Observation at 48 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: No swelling (normal) was seen in all the animals.

 

Observation at 72 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 - Blood vessels normal was observed in all the animals; Chemosis: No swelling (normal) was seen in all the 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, 0.67, 0.00; 0.00, 0.00, 0.67, 0.00 and 0.00, 0.00, 0.67, 0.00, respectively.

 

Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed at 72 hours. Hence, the test chemical was considered as “Non Irritant” to New Zealand White male rabbit eyes and being classified as “Not Classified as an Eye Irritant” as per the criteria of CLP regulation.

Justification for classification or non-classification

The skin and eye irritation potential of test chemical was observed in experimental studies. The results obtained from these studies indicate that the test chemical is not likely to cause skin and eye irritation. Hence the test chemical can be classified under the category for “Not Classified” for skin and eye as per CLP.