Registration Dossier

Diss Factsheets

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 and on day 7 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. 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, 1.67, 0.00; 0.00, 0.00, 1.00, 1.00 and 0.00, 0.00, 2.00, 1.00, respectively. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of all 3 animals was observed at day 7. Hence, the test chemical is considered as mildly irritating to the eyes of female New Zealand White rabbits, and thus comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Category 2”.

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
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 or test system and environmental conditions:
TEST ANIMALS- Source:Procured from RABBI ROOF, Hyderabad
.- Age at study initiation:4 to 5 Months (Approximately)
- Weight at study initiation: 1.874-2.272 kg
- Housing:The animals were housed individually in stainless steel cages in a controlled environment.
- Diet (e.g. ad libitum): Conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune), ad libitum
- Water (e.g. ad libitum):Aqua guard filtered tap water, ad libitum-
Acclimatization period: Rabbits were acclimatized to the test conditions for a period of 5 days (Animal No. 1) and 9 days (Animal No. 2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.30-22.50 °C
- Humidity (%): 45.00-68.40 %
- Air changes (per hr):More than 12 changes per hour
- Photoperiod (hrs dark / hrs light):12-hrs light/12-hrs dark
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 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 male 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:
animal: #1 ,#2 and #3
Time point:
other: At 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal: #1 ,#2 and #3
Time point:
other: At 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
not specified
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 1, 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method. In Animal Nos. 1, 2 and 3 at 1 hour observation after post patch removal, revealed no erythema and oedema at 1, 24, 48 and 72 hour observation. 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 all animals was comparable with pre-treatment body weight.

Table 1

Skin Reaction

 

In Treated area Dose:0.5 ml of test item                                                                                             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

Left

0

0

0

0

0

0

0

0

 

In Control areaDose: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

Left

0

0

0

0

0

0

0

0

2

Confirmatory

Left

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

2.272

2.330

2

1.874

1.912

3

1.938

1.948

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:
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 male. The hairs of all the rabbits were clipped at contralateral sites, approximately 24 hours prior to treatment. A dose of 0.5 g of test item (moistened with 0.5 ml 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, no erythema and no oedema observed at 1, 24 and 48 hour of observation hence a confirmatory test was conducted on additional two rabbits (No. 2 and 3) to confirm the non-irritant nature of the test item.

The patch was removed after 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. In animal no. 2 and 3 at 1 hour observation post patch removal, revealed no erythema and no oedema. At 24, 48 and 72 hour post patch removal, animals showed no erythema and no oedema.

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.

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
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Principles of method if other than guideline:
The objective of the study was to assess the irritant and/or corrosive effects of test chemical on eye when exposed by an ocular route in rabbits.
GLP compliance:
not specified
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS- Source:Procured from RABBI ROOF, Hyderabad
.- Age at study initiation:4 to 5 Months (Approximately)
- Weight at study initiation: 1.999-2.268 kg
- Housing:The animals were housed individually in stainless steel cages in a controlled environment.
- Diet (e.g. ad libitum): Conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune), ad libitum
- Water (e.g. ad libitum):Aqua guard filtered tap water, ad libitum-
Acclimatization period: Rabbits were acclimatized to the test conditions for a period of 5 days (Animal No. 1) and 9 days (Animal No. 2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.20-22.60°C
- Humidity (%): 42.00-74.70%
- Air changes (per hr):More than 12 changes per hour
- Photoperiod (hrs dark / hrs light):12-hrs light/12-hrs dark
Vehicle:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml
- Concentration (if solution): No data available

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

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

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

Table 1 : Individual Animal Eye Irritation Scores

 

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

Animal Numbers

1

2

3

Application Side

Right

Right

Right

Eye Reactions

At hour

At hour

At hour

*

1

24

48

72

7 days

*

1

24

48

72

7 days

*

1

24

48

72

7 days

Corneal Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Area of Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Iris

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctiva

0

1

1

2

2

0

0

1

1

1

1

0

0

1

2

2

2

0

Chemosis

0

0

0

0

0

0

0

1

1

1

1

0

0

1

1

1

1

0

Corneal Damage%

0

15

20

 

 

Dose:Untreated (Control Eye)                                                                         Sex:Female

Animal Numbers

1

2

3

Application Side

Left

Left

Left

Eye Reactions

At hour

At hour

At hour

*

1

24

48

72

Day 7

*

1

24

48

72

Day 7

*

1

24

48

72

Day 7

Corneal Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Area of Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Iris

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctiva

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Chemosis

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Corneal Damage%

0

0

0

Key:*= Pre-exposure eye examination.

 

 

 

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

            Animal No.

 Eye Reaction

1

2

3

Corneal Opacity

0.00

0.00

0.00

Iris

0.00

0.00

0.00

Conjunctiva

1.67

1.00

2.00

Chemosis

0.00

1.00

1.00

 

 

Formula :

 

Mean Eye Irritation Score =

                          

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

Number of the Observations (3)

 

Table 2: Individual AnimalClinicalSigns

 

Sex:Female

Animal No.

Days (Post application observation)

0

1

2

3

4

5

6

7

1

1

1

1

1

1

1

1

1

2

1

1

1

1

1

1

1

1

3

1

1

1

1

1

1

1

1

Key:1 = Normal

 

  

Table 3: Individual Animal Body Weight

Sex :Female

Animal No.

Animal Body Weight (kg)

Prior to application

At termination

1

2.268

2.361

2

1.999

2.201

3

2.092

2.213

Key:kg = Kilogram

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

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

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

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

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

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

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

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

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

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

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

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (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. The hairs of all the rabbits were clipped at contralateral sites, approximately 24 hours prior to treatment. A dose of 0.5 g of test item (moistened with 0.5 ml 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, no erythema and no oedema observed at 1, 24 and 48 hour of observation hence a confirmatory test was conducted on additional two rabbits (No. 2 and 3) to confirm the non-irritant nature of the test item.

The patch was removed after 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. In animal no. 2 and 3 at 1 hour observation post patch removal, revealed no erythema and no oedema. At 24, 48 and 72 hour post patch removal, animals showed no erythema and no oedema.

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.

 

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; 0.1 ml of test item was instilled in the other (treated) eye of each rabbit. The eye was observed at 1, 24, 48, 72 hour, day 7, day 14 and day 21 for animal no. 1 and 3 whereas at 1, 24, 48 and 72 hour after test item instillation. Ophthalmoscope was used for scoring of eye lesions.

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

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

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

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

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

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

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

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

 

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 irritation but can cause mild eye damage. Hence the test chemical can be classified under the category for “Not Classified” skin and “Category 2” for eye as per CLP.