Registration Dossier

Administrative data

Description of key information

Skin irritation:

The dermal irritation potential of target chemical was assessed in in-vivo experimental study.Based on the available study,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 in-vivo experimental study.Based on the available study,it can be concluded that the test chemical is able to cause eye irritation and considered as irritating. 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 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 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:4 to 5 Months (Approximately)
- Weight (Prior to Treatment):Minimum: 2.066 kg and Maximum: 2.418 kg
- Health Status : Healthy young adult 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 10 days (Animal No.-2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 19.30 °C and Maximum: 22.50 °C
- Humidity (%):Minimum: 43.80 % 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
- Concentration (if solution):N/A

VEHICLE
- Amount(s) applied (volume or weight with unit):0.5 ml 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 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.

In Animals Nos.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 weights of all Animals were comparable 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

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 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

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:Male

Animal

No.

Body Weight (kg)

Prior to Dosing

At termination

1

2.418

2.464

2

2.066

2.080

3

2.274

2.296

 

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:
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 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 male 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 gm of test item moistened with 0.5 ml of distilled water 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 in Animal No. 1, there was no erythema and oedema observed at 1, 24, 48 and 72 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 Nos. 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 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 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.

 

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:
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 Animal Facility, A. P., India.
- Age at study initiation:3.5 to 4.5 Months (Approximately)
- Weight at study initiation:Minimum: 1.576 kg and Maximum: 2.746 kg (Prior to Treatment)
- Health Status :Healthy young adult.
- 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 9 days (Animal No. 2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 19.20 °C and Maximum: 22.70 °C
- Humidity (%):Minimum: 42.00 % and Maximum: 74.70 %.
- 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 gm
- 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, day 7, 14 and 21 after instillation of test item.
Number of animals or in vitro replicates:
3 male rabbits
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done):The treated eye of rabbit was washed with normal saline.
- Time after start of exposure:24 hours

SCORING SYSTEM:Draize Method

TOOL USED TO ASSESS SCORE: Ophthalmoscope and fluorescein strips.
Irritation parameter:
cornea opacity score
Basis:
animal: #1,#2 and #3
Time point:
other: 24,48,72 hrs,7days,14 days and 21 days
Score:
0
Max. score:
4
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,7days,14 days and 21 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,7days,14 days and 21 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,7days,14 days and 21 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,Day 7 and Day 14
Score:
1
Max. score:
4
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
cornea opacity score
Basis:
animal: #1,#2 and #3
Time point:
other: Day 21
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
iris score
Basis:
animal: #1,#2 and #3
Time point:
other: After 24, 48, 72 Hours,day 7 ,day 14 and day 21
Score:
0
Max. score:
2
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: After 24 Hours
Score:
1
Max. score:
3
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
other: After 48 Hours
Score:
1.67
Max. score:
3
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: After 72 Hours
Score:
1.33
Max. score:
3
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
conjunctivae score
Basis:
animal: #1,#2
Time point:
other: Day 7 and Day 14
Score:
2
Max. score:
3
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: Day 7 and Day 14
Score:
1
Max. score:
3
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
conjunctivae score
Basis:
animal: #1,#2 and #3
Time point:
other: Day 21
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: After 24 Hours
Score:
1
Max. score:
4
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
chemosis score
Basis:
animal: #2 and #3
Time point:
other: After 48 Hours and 72 Hours
Score:
1.33
Max. score:
4
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: Day 7
Score:
2
Max. score:
4
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
chemosis score
Basis:
animal: #2,#3
Time point:
other: Day 7
Score:
1
Max. score:
4
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
chemosis score
Basis:
animal: #1 and #2
Time point:
other: Day 14
Score:
1
Max. score:
4
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: Day 14
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
other: For Treated eye
Irritation parameter:
chemosis score
Basis:
animal: #1,#2 and #3
Time point:
other: Day 21
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
other: For treated eye
Irritant / corrosive response data:
The following were observed in treated rabbits.
In the initial test, 0.1 gm 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. As animal no. 1 showed ocular lesions hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 0.1 gm 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. 1, 2 and 3 at till day 14observation which recovered at day 21.

Untreated eye of all the three rabbits was normal throughout the experimental period of 21 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 - Some blood vessels definitely hyperaemic (injected) was seen in all 3 animals; Chemosis: some swelling above normal (includes nictitating membranes) was seen in animal numbers 2 and 3 whereas animal number 1 was normal.

Observation at 24 hours after instillation of test item revealed: Cornea- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible in all 3 animals; Area of Opacity- One quarter (or less) but not zero; Iris: Normal in all 3 animals; Conjunctivae - Some blood vessels definitely hyperaemic (injected) in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) 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 30 %, 30 % and 20 % damage in animal no. 1, 2 and 3, resectively.

Observation at 48 hours after instillation of test item revealed: Cornea- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible in all 3 animals; Area of Opacity- One quarter (or less) but not zero; Iris: Normal in all 3 animals; Conjunctivae - Some blood vessels definitely hyperaemic (injected) was seen in animal number 1 whereas diffuse, crimson color; individual vessels not easily discernible in animal number 2 and 3; Chemosis: Obvious swelling with partial eversion of lids was seen in animal numbers. 2 and 3 whereas some swelling above normal (includes nictitating membranes) was seen in animal no.1.

Observation at 72 hours after instillation of test item revealed: Cornea- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible in all 3 animals; Area of Opacity- One quarter (or less) but not zero; Iris: Normal in all 3 animals; Conjunctivae - Some blood vessels definitely hyperaemic (injected) was seen in animal numbers 1 and 3 whereas diffuse, crimson color; individual vessels not easily discernible in animal number 2; Chemosis: some swelling above normal (includes nictitating membranes) was noticed in all the animals.

Observation at day 7 after instillation of test item revealed: Cornea- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible in all 3 animals; Area of Opacity- One quarter (or less) but not zero; Iris: Normal in all 3 animals; Conjunctivae - Some blood vessels definitely hyperaemic (injected) was seen in animal number 3 whereas diffuse, crimson color; individual vessels not easily discernible in animal numbers 1 and 2; Chemosis: some swelling above normal (includes nictitating membranes) was noticed in animal numbers 2 and 3 whereas obvious swelling with partial eversion of lids was seen in animal number 1.

Observation at day 14 after instillation of test item revealed: Cornea- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible in animal numbers 1 and 2 whereas animal number 3 recovered to normal; Area of Opacity- One quarter (or less) but not zero in animal numbers 1 and 2 whereas animal number 3 recovered to normal; Iris: Normal in all 3 animals; Conjunctivae - Some blood vessels definitely hyperaemic (injected) was seen in animal number 3 whereas diffuse, crimson color; individual vessels not easily discernible in animal numbers 1 and 2; Chemosis: some swelling above normal (includes nictitating membranes) was noticed in animal numbers 1 and 2 whereas animal number 3 recovered to normal.

Observation at day 21 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: recovered to 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 1.00, 0.00, 1.00, 1.00; 1.00, 0.00, 1.67, 1.33 and 1.00, 0.00, 1.33,1.33, 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
Animal Number 1 revealed marginal decrease in body weight where as Animal Numbers 2 and 3 were observed with marginally body weight gain on day 14 as compared to day 0.

Table 1 : Individual Animal Eye Irritation Scores

 

In Treated area Dose:0.1 gm of test item                                                             Sex:Male

Animal Numbers

1

2

3

Application Side

Right

Right

Right

Eye Reactions

At hour

At hour

At hour

*

1

24

48

72

Day 7

Day 14

Day 21

*

1

24

48

72

Day 7

Day 14

Day 21

*

1

24

48

72

Day 7

Day 14

Day 21

Corneal Opacity

0

0

1

1

1

1

1

0

0

0

1

1

1

1

1

0

0

0

1

1

1

1

1

0

Area of Opacity

0

0

1

1

1

1

1

0

0

0

1

1

1

1

1

0

0

0

1

1

1

1

1

0

Iris

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctiva

0

1

1

1

1

2

2

0

0

1

1

2

2

2

2

0

0

1

1

2

1

1

1

0

Chemosis

0

0

1

1

1

2

1

0

0

1

1

2

1

1

1

0

0

1

1

2

1

1

0

0

Corneal Damage%

30

30

20

 

 

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

Day 7

Day 14

Day 21

*

1

24

48

72

Day 7

Day 14

Day 21

*

1

24

48

72

Day 7

Day 14

Day 21

Corneal Opacity

0

0

0

0

0

0

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

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

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

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

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

1.00

1.00

1.00

Iris

0.00

0.00

0.00

Conjunctiva

1.00

1.67

1.33

Chemosis

1.00

1.33

1.33

 

 

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

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

2

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

3

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

Key:1 = Normal

 

  

Table 3: Individual Animal Body Weight

Sex :Male

Animal No.

Animal Body Weight (kg)

Prior to application

At termination

1

1.576

1.506

2

1.746

1.891

3

1.686

1.799

Key:kg = Kilogram

 


Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of all 3 animals was observed at day 21.Hence, test chemical is “Irritant” to New Zealand White Female rabbit eyes
and is thus 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 gm of test item was instilled in the other (treated) eye of each rabbit.The eye was observed at 1, 24, 48, 72 hours, day 7, 14 and 21 after test item instillation.Ophthalmoscope was used for scoring of eye lesions. In the initial test,0.1 gm 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. As animal no. 1 showed ocular lesions, hence confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 0.1 gmof test itemwas 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. 1, 2 and 3 till day 14 observations which recovered at day 21. Untreated eye of all the three rabbits was normal throughout the experimental period of 14 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 -Some blood vessels definitely hyperaemic (injected) was seen in all 3 animals;Chemosis:some swelling above normal (includes nictitating membranes) was seen in animal numbers 2 and 3 whereas animal number 1 was normal. Observation at 24 hours after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visiblein all 3 animals; Area of Opacity-One quarter (or less) but not zero; Iris: Normal in all 3 animals;Conjunctivae-Some blood vessels definitely hyperaemic (injected) in all the animals;Chemosis:Some swelling above normal (includes nictitating membranes) 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 30 %, 30 % and 20 % damage in animal no. 1, 2 and 3, resectively.Observation at 48 hours after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visiblein all 3 animals; Area of Opacity-One quarter (or less) but not zero; Iris: Normal in all 3 animals; Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal number 1 whereas diffuse, crimson color; individual vessels not easily discernible in animal number 2 and 3;Chemosis:Obvious swelling with partial eversion of lids was seen in animal numbers. 2 and 3 whereas some swelling above normal (includes nictitating membranes) was seen in animal no.1. Observation at 72 hours after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visiblein all 3 animals; Area of Opacity-One quarter (or less) but not zero; Iris: Normal in all 3 animals; Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal numbers 1 and 3 whereas diffuse, crimson color; individual vessels not easily discernible in animal number 2;Chemosis:some swelling above normal (includes nictitating membranes) was noticed in all the animals. Observation at day 7 after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visiblein all 3 animals; Area of Opacity-One quarter (or less) but not zero; Iris: Normal in all 3 animals; Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal number 3 whereas diffuse, crimson color; individual vessels not easily discernible in animal numbers 1 and 2;Chemosis:some swelling above normal (includes nictitating membranes) was noticed in animal numbers 2 and 3 whereas obvious swelling with partial eversion of lids was seen in animal number 1. Observation at day 14 after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visiblein animal numbers 1 and 2 whereas animal number 3 recovered to normal; Area of Opacity-One quarter (or less) but not zeroin animal numbers 1 and 2 whereas animal number 3 recovered to normal; Iris: Normal in all 3 animals; Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal number 3 whereas diffuse, crimson color; individual vessels not easily discernible in animal numbers 1 and 2;Chemosis:some swelling above normal (includes nictitating membranes) was noticed in animal numbers 1 and 2 whereas animal number 3 recovered to normal. Observation at day 21 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:recovered to 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 1.00, 0.00, 1.00, 1.00; 1.00, 0.00, 1.67, 1.33 and 1.00, 0.00, 1.33,1.33, respectively. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of all 3 animals was observed at day 21. Hence, test chemical is “Irritant” to New Zealand White Female rabbit eyes and is thus classified as a eye irritant.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

The test chemical has been investigated for potential for dermal irritation to a greater or lesser extent. The study is based on in-vivo experiment conducted in 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 male 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 gm of test item moistened with 0.5 ml of distilled water 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 in Animal No. 1, there was no erythema and oedema observed at 1, 24, 48 and 72 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 Nos. 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 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 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.

The above study leads 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 - not irritant”.

Eye Irritation:

The test chemical has been investigated for potential for ocular irritation to a greater or lesser extent. The study is based on in-vivo experiment conducted in rabbits 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 gm of test item was instilled in the other (treated) eye of each rabbit.The eye was observed at 1, 24, 48, 72 hours, day 7, 14 and 21 after test item instillation.Ophthalmoscope was used for scoring of eye lesions. In the initial test,0.1 gm 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. As animal no. 1 showed ocular lesions, hence confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 0.1 gmof test itemwas 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. 1, 2 and 3 till day 14 observations which recovered at day 21. Untreated eye of all the three rabbits was normal throughout the experimental period of 14 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 -Some blood vessels definitely hyperaemic (injected) was seen in all 3 animals;Chemosis:some swelling above normal (includes nictitating membranes) was seen in animal numbers 2 and 3 whereas animal number 1 was normal. Observation at 24 hours after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visiblein all 3 animals; Area of Opacity-One quarter (or less) but not zero; Iris: Normal in all 3 animals;Conjunctivae-Some blood vessels definitely hyperaemic (injected) in all the animals;Chemosis:Some swelling above normal (includes nictitating membranes) 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 30 %, 30 % and 20 % damage in animal no. 1, 2 and 3, resectively.Observation at 48 hours after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visiblein all 3 animals; Area of Opacity-One quarter (or less) but not zero; Iris: Normal in all 3 animals; Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal number 1 whereas diffuse, crimson color; individual vessels not easily discernible in animal number 2 and 3;Chemosis:Obvious swelling with partial eversion of lids was seen in animal numbers. 2 and 3 whereas some swelling above normal (includes nictitating membranes) was seen in animal no.1. Observation at 72 hours after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visiblein all 3 animals; Area of Opacity-One quarter (or less) but not zero; Iris: Normal in all 3 animals; Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal numbers 1 and 3 whereas diffuse, crimson color; individual vessels not easily discernible in animal number 2;Chemosis:some swelling above normal (includes nictitating membranes) was noticed in all the animals. Observation at day 7 after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visiblein all 3 animals; Area of Opacity-One quarter (or less) but not zero; Iris: Normal in all 3 animals; Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal number 3 whereas diffuse, crimson color; individual vessels not easily discernible in animal numbers 1 and 2;Chemosis:some swelling above normal (includes nictitating membranes) was noticed in animal numbers 2 and 3 whereas obvious swelling with partial eversion of lids was seen in animal number 1. Observation at day 14 after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visiblein animal numbers 1 and 2 whereas animal number 3 recovered to normal; Area of Opacity-One quarter (or less) but not zeroin animal numbers 1 and 2 whereas animal number 3 recovered to normal; Iris: Normal in all 3 animals; Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal number 3 whereas diffuse, crimson color; individual vessels not easily discernible in animal numbers 1 and 2;Chemosis:some swelling above normal (includes nictitating membranes) was noticed in animal numbers 1 and 2 whereas animal number 3 recovered to normal. Observation at day 21 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:recovered to 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 1.00, 0.00, 1.00, 1.00; 1.00, 0.00, 1.67, 1.33 and 1.00, 0.00, 1.33,1.33, respectively. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of all 3 animals was observed at day 21. Hence, test chemical is “Irritant” to New Zealand White Female rabbit eyes and is thus classified as a eye irritant.

The above study leads to a conclusion that Test chemical is indeed irritating to eye. Hence, comparing the above annotations with the criteria of CLP regulation, Test chemical can be classified under the category “category 2 - irritant”.

Justification for classification or non-classification

The skin and eye irritation potential of test chemical was observed in experimental study. The results obtained from these studies indicate that the chemical is not likely to cause skin irritation while irritant to eye. Hence the test chemical can be classified under the category “Not Classified” for skin irritation and “Category 2” for eye as per CLP.