Registration Dossier

Administrative data

Description of key information

Skin irritation:

The dermal irritation potential of target chemical was assessed in various in-vivo experimental studies.Based on the available studies,it can be concluded that the test chemical is unable to cause skin irritation and considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified''.

Eye irritation:

The ocular irritation potential of target chemical was assessed in various in-vivo experimental studies.Based on the available studies,it can be concluded that the test chemical is unable to cause eye irritation and considered as not irritating. 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 study report
Qualifier:
according to guideline
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 or test system and environmental conditions:
TEST ANIMALS
- Source:Procured from LIVEON BIOLABS PVT. LTD.
- Age at study initiation:3 to 3.5 Months (Approximately)
- Weight (Prior to Treatment):Minimum: 1.956 kg and Maximum: 2.398 kg (Prior to Treatment)
- Health Status : Healthy young adults rabbits were used for the study. 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 8 days (Animal No.-1) and 10 days (Animal No.-2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 19.90 °C Maximum: 22.30 °C
- Humidity (%):Minimum: 52.80 % Maximum: 67.40 %
- 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:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):0.5 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:
4 hours
Observation period:
72 hours
Number of animals:
3 female 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:
0.33
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 following were observed in treated rabbits.
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.
Animal No.1 revealed Very slight erythema (barely perceptible) and no edema at 1 & 24 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. Animal no. 1 recovered to normal at 48 hours observation period.
After 4 hours of exposure, animal no. 2 and 3 revealed very slight erythema (barely perceptible) and no edema at 1 & 24 hours observation. Animal no. 2 and 3 recovered to normal at 48 hours observation period.
The individual mean score at 24, 48 and 72 hours for animal nos. 1, 2 and 3 were 0.33, 0.33, 0.33 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.


 

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

Left

1

1

0

0

0

0

0

0

2

Confirmatory

Right

1

1

0

0

0

0

0

0

3

Right

1

1

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

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

Key:h = Hour.

 

Erythema                                                                                                       Oedema

0 =No erythema                                                                                          0 =No oedema

1 = Very slight erythema (barely perceptible)
Table 1 Continued…

Mean Individual Animal Score at 24, 48 and 72 hours

 

                     Animal Number                  

Observations                      

1

2

3

Erythema

0.33

0.33

0.33

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

1.962

2.042

2

1.956

1.988

3

2.398

2.470

 

 

Table 3

Individual AnimalClinicalSigns

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 0.33, 0.33, 0.33 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 Female 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 in Rabbits, . This study was performed as per OECD guideline No. 404. Three healthy young adult female rabbits were used for conducting acute dermal irritation study.Body weights were re­corded on day 0 (prior to application) and at termination.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 of0.5 ml of test item (as such) wasappliedto the skin,over an area of approximately 6 x 6 cm clippedof haironone 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, animal no. 1 revealed very slight erythema (barely perceptible)and no edema at 1, 24 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. Animal no. 1 recovered to normal at 48 hours observation.After 4 hours of exposure, animal no. 2 and 3 revealed very slight erythema (barely perceptible)and no edema at 1, 24 hours observation which recovered to 48 hours observation.The patch was removed after 4 hours and animals wereobservedfor erythema and oedemaat 1, 24, 48 and 72 hours after patch removal, evaluatedand graded as per draize method. The individual mean score at 24, 48 and 72 hoursfor animal nos. 1, 2 and 3 were 0.33, 0.33, 0.33 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 Female New Zealand White rabbits under the experimental conditions tested and is thus not classified as a skin irritant.

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 report
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 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 LIVEON BIOLABS PVT. LTD.
- Age at study initiation:2 to 3.5 Months (Approximately)
- Weight (Prior to Treatment):Minimum: 2.048 kg and Maximum: 2.168 kg (Prior to Treatment)
- Health Status : Healthy young adults rabbits were used for the study. 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 7 days (Animal No.-1) and 10 days (Animal No.-2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 20.10 °C Maximum: 22.20 °C
- Humidity (%):Minimum: 57.20 % Maximum: 69.20 %
- 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):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, 72 hours and at day7 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 fluorescent strips.
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:
3
Reversibility:
not specified
Remarks on result:
other: 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:
other: 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:
other: 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:
other: 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:
other: 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:
other: Treated eye
Irritation parameter:
conjunctivae score
Basis:
animal: #1,#2 and #3
Time point:
other: 24, 48, 72 Hours
Score:
0
Max. score:
2
Reversibility:
not specified
Remarks on result:
other: Treated eye
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:
other: 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 the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) in all the animals; Chemosis: No swelling (Normal) were 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 - Blood vessels recovered to normal 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 %, 0% and 0% damage in Animal Nos 1, 2 and 3, respectively.
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 (normal) was observed in all the Animals.
Untreated eye of the treated rabbits was normal throughout the experimental period of 72 hours.
The individual mean score for Animal Nos. 1, 2 and 3 at 24, 48, 72 hours for Corneal opacity, iris, conjunctiva, chemosis were found 0.00, 0.00, 0.00, 0.00 ; 0.00, 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, 0.00, 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
All rabbits were weighed on test day 0 (prior to application) and at termination. The body weights increased in all the animals as compared with pre-treatment body weight

Table 1 : Individual Animal Eye Irritation Scores

 

In Treated area Dose:100 mg 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

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

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

1

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

0.00

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

Sex :Female

Animal No.

Animal Body Weight (kg)

Prior to application

At termination

1

2.186

2.246

2

2.048

2.148

3

2.132

2.196

Key:kg = Kilogram

Interpretation of results:
not irritating
Conclusions:
Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbit no. 1, 2 and 3 was observed till 72 hours which were recovered on day 7.Hence, test chemical is “Non Irritant” to New Zealand White Female rabbit eyes and is thus not classified as a eye irritant.
Executive summary:

Acute Eye Irritation/Corrosion Study of test chemical in Rabbits. This study was performed as per OECD guideline No. 405. 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 (as such)was instilled in the other (treated) eye of rabbits.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 itemwas applied into the conjunctival sac of the right eye of Animal No.1. The left eye of the rabbit served as the control. As no severe ocular lesions were observed till 24 hour observation period, hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3);0.1 ml of test itemwas instilled into the conjunctival sac of right eye and left eye served as the control. Ocular lesions were observed at 1 hour in animal number 2 and 3. Untreated eye of the treated 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) in all the animals;Chemosis:No swelling (Normal) were 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 -Blood vessels recovered to normal 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 %, 0% and 0% damage in animal nos. 1, 2 and 3, respectively. 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 (normal) 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, chemosis were found 0.00, 0.00, 0.00, 0.00 ; 0.00, 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, 0.00, respectively. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbit no. 1, 2 and 3 was observed till 72 hours which were recovered on day 7.Hence, test chemical is “Non Irritant” to New Zealand White Female rabbit eyes and is thus not classified as a eye irritant.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Irritation:

In different studies, the test chemical has been investigated for potential for dermal irritation to a greater or lesser extent. The studies are based on in-vivo experimental conducted in human and rabbits which have been summarized as below:

Acute Dermal Irritation/corrosion Study of test chemical in Rabbits, . This study was performed as per OECD guideline No. 404. Three healthy young adult female rabbits were used for conducting acute dermal irritation study.Body weights were re­corded on day 0 (prior to application) and at termination.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 of0.5 ml of test item (as such) wasappliedto the skin,over an area of approximately 6 x 6 cm clippedof haironone 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, animal no. 1 revealed very slight erythema (barely perceptible)and no edema at 1, 24 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. Animal no. 1 recovered to normal at 48 hours observation.After 4 hours of exposure, animal no. 2 and 3 revealed very slight erythema (barely perceptible)and no edema at 1, 24 hours observation which recovered to 48 hours observation.The patch was removed after 4 hours and animals wereobservedfor erythema and oedemaat 1, 24, 48 and 72 hours after patch removal, evaluatedand graded as per draize method. The individual mean score at 24, 48 and 72 hoursfor animal nos. 1, 2 and 3 were 0.33, 0.33, 0.33 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 Female New Zealand White rabbits under the experimental conditions tested and is thus not classified as a skin irritant.

 

The above studies were supported by ,a dermal irritation study which was conducted on New Zealand white rabbits in accordance with OECD 404 to assess the irritation parameter of the test chemical.In the initial test one healthy rabbit of body weight 2.14 kg was selected for the study after acclimatization.The test compound in the amount of 0.5 ml was applied at the different sites on the shaven back skin of animal. The hairs of back sides were removed (approximately 6 cm2) one day earlier before the treatment.The site of application was covered with a imperviousdressing securedin position with adhesive tape.The first patch was applied on the shaven back skin of rabbit and removed after three minutes. No serious reaction was observed at the site of application. The second patch was applied on the different shaven back side and removed after one hour. There were no signs of skin reaction observed at this site of application. Finally, a third patch was applied at a different site and was removed after four hour.No reaction was observed after four hour patch removal .Finally, the animal was observed for 14 days, for any irritation and corrosion.Because no corrosive effect observed in the initial test, a confirmatory test was done in order to confirm the irritant or negative response of the test substance by using two additional animals.In the confirmatory test the test compound in the amount of 0.5 ml was applied on the shaven back skin of two animals, each with one patch, for an exposure period of four hours.After four hours the patch was removed and the skin reactions were graded according to draize's method.The test compound when applied at the dose level of 0.5 gm on shaven back skin (approximately 6 cm2) of rabbit did not produced any dermal irritation in terms of inflammatory changes after 4 hours patch test removal. However, there was no corrosivity of the test compound was observed during the entire observation period of 14 days.The dermal irritation index of the test chemical in New Zealand white rabbits was calculated as 0.00 and test compound can be classified under "Not Classified".

Both the above studies were further supported by Acute Dermal Irritation/corrosion Study,in which 6 rabbits were used . The undiluted test chemical was applied dermally to the skin of rabbits at a dose of 20 ml/kg bw for 4 hours under 14 days observation period. Since no signs of irritation were detected neither during contact with the substances nor 24 hours later, it was considered as not irritating to skin after 4 hours exposure period.

All these studies lead to a conclusion that Test chemical is indeed not irritating to skin. Hence, comparing the above annotations with the criteria of CLP regulation, Test chemical can be classified under the category “Not Classified”.

 

Eye Irritation:

In different studies, the test chemical has been investigated for potential for ocular irritation to a greater or lesser extent. The studies are based on in-vivo experimental conducted in rabbits conducted which have been summarized as below:

Acute Eye Irritation/Corrosion Study of test chemical in Rabbits. This study was performed as per OECD guideline No. 405. 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 (as such)was instilled in the other (treated) eye of rabbits.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 itemwas applied into the conjunctival sac of the right eye of Animal No.1. The left eye of the rabbit served as the control. As no severe ocular lesions were observed till 24 hour observation period, hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3);0.1 ml of test itemwas instilled into the conjunctival sac of right eye and left eye served as the control. Ocular lesions were observed at 1 hour in animal number 2 and 3. Untreated eye of the treated 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) in all the animals;Chemosis:No swelling (Normal) were 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 -Blood vessels recovered to normal 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 %, 0% and 0% damage in animal nos. 1, 2 and 3, respectively. 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 (normal) 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, chemosis were found 0.00, 0.00, 0.00, 0.00 ; 0.00, 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, 0.00, respectively. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbit no. 1, 2 and 3 was observed till 72 hours which were recovered on day 7.Hence, test chemical is “Non Irritant” to New Zealand White Female rabbit eyes and is thus not classified as a eye irritant.

In addition to another study, the acute eye irritation study of the test chemical was conducted in New Zealand White Rabbit as per the OECD- 405 Guideline. 3 female young adult New Zealand White rabbits were used for the study.Both eyes of each experimental animal provisionally selected and examined for any eye irritation, ocular defects, and pre-existing corneal injury within 24 hours before testing of compound. The test chemical in the amount of 0.1 ml placed in the conjunctival sac of one eye of each animal after gentle pulling the lower eye lid away from the eye ball. The eye lids were then gently held together for about one second to prevent the loss of the material. The other eyes which remains untreated, served as control.The eyes of the test animal did not washed for at least 24 hours following instillation of the test compound. At 24 hours a washout may be used if considered appropriate. In the confirmatory test, the test compound when applied to the conjunctival sac of the rabbits did not produced any eye irritation or inflammation and any discharge after application of test compound. Furthermore, no other clinical signs of toxicity such as pain, distress etc. were not recorded upto the entire observation period of 21 days.Based on above findings, it can be concluded that the test compound was not irritating when applied the amount of 0.1 ml in the conjunctival sac of the rabbits under test condition.

Both the above studies are further supported by acute Eye Irritation/Corrosion Study,in which 0.1 ml test chemical applied to rabbit's eye for 24 hours of exposure period. Since the test chemical did not caused eye lesions, it was considered as not irritating to animal eye.

The above studies were further supported by Acute Eye Irritation/Corrosion Study,in which rabbit's eye were treated with test chemical at dose concentration of 20%.The test chemical was applied on the mucous membrane of the rabbit eyelid which was given in the form of capillary injection. Since the test chemical did not showed any corneal injuries, it was considered as  not irritating to animal eye.

All these studies lead to a conclusion that Test chemical is indeed not irritating to eye. Hence, comparing the above annotations with the criteria of CLP regulation, Test chemical can be classified under the category “Not Classified”.

Justification for classification or non-classification

The skin and eye irritation potential of test chemical was observed in various 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 “Not Classified” for skin and eye as per CLP.