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 dose of 0.5 gm of test item moistened with 0.5 ml of distilled water 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 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”.

Eye Irritation:

Acute Eye Irritation/Corrosion Study of Test chemical in Rabbits, was performed as per OECD guideline No. 405. The 100 mg of test item was instilled in the other (treated) eye of each rabbit. The eye was observed at 1, 24, 48 and 72 hour 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, 0.00, 0.00; 0.00, 0.00, 0.00, 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 rabbit no. 1 and 2 was observed at 24 hours and rabbit no. 3 at 48 hours. Hence, the test chemical is considered as Non-Irritating to the eyes of female New Zealand White rabbits under the experimental conditions tested and Classified as “Category- Not Classified” as per CLP Regulation.

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 study
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 in Rabbits.
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:Procured from RABBI ROOF, Hyderabad
- Age at study initiation:3.5 to 4.5 Months (Approximately)
- Weight (Prior to Treatment):Minimum: 2.049 kg and Maximum: 2.336 kg
- Health Status : Healthy young adults rabbits 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 5 days (Animal No.-1) and 8 days (Animal No.-2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 20.00 °C and Maximum: 22.50 °C
- Humidity (%):Minimum: 47.20 % and Maximum: 68.40 %
- Air changes (per hr):More than 12 changes per hour
- Photoperiod (hrs dark / hrs light):12:12

Type of coverage:
occlusive
Preparation of test site:
clipped
Vehicle:
other: distilled water
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):0.5 gm

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

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

SCORING SYSTEM:Draize Method
Irritation parameter:
erythema score
Basis:
animal: #1 ,#2 and #3
Time point:
other: At 24, 48 and 72 hours
Score:
0
Max. score:
3
Reversibility:
no data
Remarks on result:
other: not irritating
Irritation parameter:
edema score
Basis:
animal: #1 ,#2 and #3
Time point:
other: At 24, 48 and 72 hours
Score:
0
Max. score:
3
Reversibility:
no data
Remarks on result:
other: not irritating
Irritant / corrosive response data:
The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method.

In Animals No.1, 2 and 3 after post patch removal, revealed no erythema and oedema at 24 and 48 and 72 hour during the observation period.

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.
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.

Body weight
Body weight of Animal No. 1 was marginally decreased and in Animal No. 2 and 3 it was increase as compared with pre-treatment body weight.

Table 1

Skin Reaction

In Treated area Dose:0.5 gm of test item                                                                                          Sex:Male

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

Left

0

0

0

0

0

0

0

0

3

Left

0

0

0

0

0

0

0

0

 

In Control area                

 Dose:0.5 ml of distilled water                              Sex:Male

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

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:Male

Animal

No.

Body Weight (kg)

Prior to Dosing

At termination

1

2.049

1.954

2

2.178

2.378

3

2.336

2.396

 

Table 3

Individual AnimalClinicalSigns

Sex:Male

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: 1 = Normal.

Interpretation of results:
other: not irritating
Conclusions:
No erythema and oedema (skin irritation) were found at the end of 72 hour observation period after patch removal.
Hence, under the experimental test conditions it was concluded that the test chemical was Non-Irritating to the skin of Male New Zealand White rabbits under the experimental conditions tested.
Executive summary:

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

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 gm of test item moistened with 0.5 ml of distilled water 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 and0.5 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 in Animal No. 1, there was no erythema and oedema observed at 1, 24 and 48 hours observation. Hence the confirmatory test was conducted on additional two rabbits (No. 2 and 3)to confirm the non irritant nature of the test item.

 

In Animals No. 2 and 3 after post patch removal, revealed no erythema and oedema at 1, 24, 48 and 72 hours observation.

The patch was removed after 4 hours and rabbits 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, under the experimental test condition it was concluded that the test chemical was Non-Irritating to the skin of Male New Zealand White rabbits under the experimental conditions tested and Classified as “Category- Not Classified” as per CLP Regulation.

 

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 the ocular route in rabbits.
GLP compliance:
yes
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:4.5 to 5.5 Months (Approximately)
- Weight at study initiation:Minimum: 2.036 kg and Maximum: 2.408 kg (Prior to Treatment)
- Health Status :Healthy young adult, Females were nulliparous and non pregnant.
- 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: 19.50 °C and Maximum: 22.20 °C
- Humidity (%)::Minimum: 41.20 % and Maximum: 59.20 %
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light):: 12:12
Vehicle:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):100 mg
- 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.
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 hours

SCORING SYSTEM:Draize Method

TOOL USED TO ASSESS SCORE: Ophthalmoscope and fluorescein strips.
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:
2
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eye
Irritation parameter:
conjunctivae score
Basis:
animal: #1 and #2
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 #3
Time point:
24/48/72 h
Score:
0.33
Max. score:
3
Reversibility:
fully reversible
Remarks on result:
no indication of irritation
Remarks:
treated group
Irritation parameter:
chemosis 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
Irritant / corrosive response data:
The following were observed in treated 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 the animals; Iris: Normal in all the animals. Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible in animal no. 1 and some blood vessels definitely hyperaemic (injected) animal no. 2 whereas in animal no. 3 blood vessels were normal; Chemosis: Some swelling above normal (includes nictitating membranes) in animal no. 1 whereas no swelling was observed 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 the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in animal no.3 and normal in Animal Numbers 1 and 2; Chemosis: No swelling was observed in all the 3 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 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 were normal in all the animals; Chemosis: No swelling was observed 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, 0.00, 0.00; 0.00, 0.00, 0.00, 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
All rabbits were weighed on test day 0 (prior to application) and at termination.

Table 1 : Individual Animal Eye Irritation Scores

 

In Treated area Dose:100 mg (0.1 gm) 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

*

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

2

0

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctiva

0

1

0

0

0

0

1

0

0

0

0

0

1

0

0

Chemosis

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Corneal Damage%

0

0

0

 

 

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

*

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

0.00

0.00

0.33

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:Female

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 :Female

Animal No.

Animal Body Weight (kg)

Prior to application

At termination

1

2.340

2.432

2

2.408

2.508

3

2.036

2.048

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 within 72 hours.
Hence, the test chemical is “Non Irritant” to New Zealand White Female rabbit eyes and is being classified as “Not Classified” as per the 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 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; 100 mg of test item was 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,100 mg of test item (Pulverized Form) 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. Ocular lesions were seen in animal no.1 at 1 hour observation which recovered at 24 hour hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 100 mg 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 at 1 hour observation which was recover at 24 hour and in animal no.3 at 24 hour which was recover at 48 hour 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 3 animals; Area of Opacity-Zero in all the animals;Iris:Normal in all the animals.Conjunctivae -Diffuse, crimson color; individual vessels not easily discerniblein animal no. 1 and some blood vessels definitely hyperaemic (injected) animal no. 2 whereas in animal no. 3 blood vessels were normal;Chemosis:Some swelling above normal (includes nictitating membranes) in animal no. 1 whereas no swelling was observed 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 the animals;Iris:Normal in all the animals.Conjunctivae -Some blood vessels definitely hyperaemic (injected) was observed in animal no.3 and normal in Animal Numbers 1 and 2;Chemosis:No swelling was observed in all the 3 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 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 were normal in all the animals;Chemosis:No swelling was observed 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, 0.00, 0.00; 0.00, 0.00, 0.00, 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 rabbit no. 1 and 2 was observed at 24 hours and rabbit no. 3 at 48 hours. Hence, the test chemical is considered as Non-Irritating to the eyes of female New Zealand White rabbits under the experimental conditions tested and Classified as “Category- Not Classified” as per 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 male.

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 gm of test item moistened with 0.5 ml of distilled water 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 and0.5 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 in Animal No. 1, there was no erythema and oedema observed at 1, 24 and 48 hours observation. Hence the confirmatory test was conducted on additional two rabbits (No. 2 and 3) to confirm the non-irritant nature of the test item.

 

In Animals No. 2 and 3 after post patch removal, revealed no erythema and oedema at 1, 24, 48 and 72 hours observation.

The patch was removed after 4 hours and rabbits 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, under the experimental test condition it was concluded that the test chemical was Non-Irritating to the skin of Male New Zealand White rabbits under the experimental conditions tested and Classified as “Category- Not Classified” as per CLP Regulation.

 

 

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 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; 100 mg of test item was 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,100 mg of test item (Pulverized Form) 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. Ocular lesions were seen in animal no.1 at 1 hour observation which recovered at 24 hour hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 100 mg 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 at 1 hour observation which was recover at 24 hour and in animal no.3 at 24 hour which was recover at 48 hour 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 3 animals; Area of Opacity-Zero in all the animals;Iris:Normal in all the animals. Conjunctivae -Diffuse, crimson color; individual vessels not easily discerniblein animal no. 1 and some blood vessels definitely hyperaemic (injected) animal no. 2 whereas in animal no. 3 blood vessels were normal;Chemosis:Some swelling above normal (includes nictitating membranes) in animal no. 1 whereas no swelling was observed 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 the animals;Iris:Normal in all the animals. Conjunctivae -Some blood vessels definitely hyperaemic (injected) was observed in animal no.3 and normal in Animal Numbers 1 and 2;Chemosis:No swelling was observed in all the 3 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 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 were normal in all the animals;Chemosis:No swelling was observed in all the animals.

 

The individual mean score for animal nos. 1, 2 and 3at 24, 48, 72 hours for Corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.00, 0.00; 0.00, 0.00, 0.00, 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 rabbit no. 1 and 2 was observed at 24 hours and rabbit no. 3 at 48 hours. Hence, the test chemical is considered as Non-Irritating to the eyes of female New Zealand White rabbits under the experimental conditions tested and Classified as “Category- Not Classified” as per 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 chemical is not likely to cause skin and eye irritation. Hence the test chemical can be classified under the category “Not Classified” for skin and eye as per CLP.