Registration Dossier

Administrative data

Description of key information

Skin Irritation:

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 and being classified as “Not Classified” as per CLP Classification.

Eye Irritation:

The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours (at treated site)for corneal opacity, iris, conjunctiva and chemosis were determined to be 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively.

Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed within 72 hours. 

Hence, it was concluded that the test chemical was Non-Irritating to the eyes of Male New Zealand White rabbits under the experimental conditions tested and is Classified as “Category- Not Classified" as per CLP criteria for Classification.

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 reports
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 the test chemical after topical 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 GENTOX BIOSERVICES PVT. LTD., HYDERABAD, India.
- Age at study initiation:4.0 to 5.5 Months (Approximately)
- Weight (Prior to Treatment):Minimum: 1.792 kg & Maximum: 2.138 g
- Health Status : Healthy young adult 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. Batch No.: 200009.
- 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 10 days (Animal No. 1) and 13 days (Animal No. 2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 19.00 °C and Maximum: 21.40 °C
- Humidity (%):Minimum: 41.90% and Maximum: 65.80%
- 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:
not specified
Controls:
yes, concurrent vehicle
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 (Oryctolagus cuniculus)
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
Remarks:
:For treated area dose
Basis:
animal: #1, #2 and #3
Time point:
other: At 1, 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Remarks:
:For treated area dose
Basis:
animal: #1 ,#2 and #3
Time point:
other: At 1, 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Remarks:
:For Control area Dose
Basis:
animal: #1 ,#2 and #3
Time point:
other: At 1, 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Remarks:
:For Control area Dose
Basis:
animal: #1 ,#2 and #3
Time point:
other: At 1, 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.

After 4 hours of exposure in Animal No. 1, no erythema and oedema was observed at 1 hour of observation. At 24, 48 and 72 hours observation no erythema and no oedema was observed

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 gradedas per draize method.

In Animal No. 2 & 3, at 1, 24, 48 and 72 hours, Animal Nos. 2 and 3 showed no erythema and no oedema.
The other untreated side revealed no erythema and no oedema and was found to be normal throughout the experimental 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 weights: Increase in body weight at terminal sacrifice as compared to day 0 in all the three animals

Table 1

Skin Reaction

 

In Treated area Dose:0.5 ml (as such, undiluted)                                                Sex:Female

 

Animal

No.

Test

Treated

 area

Erythema score

Oedema score

1h

24h

48h

72h

1h

24h

48h

72h

1

Initial

Right

0

0

0

0

0

0

0

0

2

Confirmatory

Left

0

0

0

0

0

0

0

0

3

Left

0

0

0

0

0

0

0

0

 

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

 

Animal

No.

Test

Treated area

Erythema score

Oedema score

1h

24h

48h

72h

1h

24h

48h

72h

1

Initial

Left

0

0

0

0

0

0

0

0

2

Confirmatory

Right

0

0

0

0

0

0

0

0

3

Right

0

0

0

0

0

0

0

0

Key: h = Hour.

Erythema                                                                                                       Oedema

0 =No erythema                                                                                           0 =No oedema

Mean Individual Animal Score at 24, 48 and 72 hours (Treated Site)

 

 

                     Animal Number                  

Observations                      

1

2

3

Erythema

0.00

0.00

0.00

Oedema

0.00

0.00

0.00

 

Table 2

Individual Animal BodyWeight

Sex:Female

Animal

No.

Body Weight (kilogram)

Prior to Dosing

At termination

1

1.792

1.866

2

2.138

2.180

3

1.920

2.020

 

Table 3

Individual Animal Clinical Signs

Sex:Female

Animal

No.

Days (Post dosing Observation)

0

1

2

3

1

1

1

1

1

2

1

1

1

1

3

1

1

1

1

Key: 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, it was concluded that the test chemical was Non-Irritating to the skin of male New Zealand White rabbits under the experimental conditions tested and being classified as “Not
Classified” as per CLP Classification.
Executive summary:

Acute Dermal Irritation/corrosion Study of the test chemical was performed on Rabbits as per OECD Guideline No. 404.

Three healthy young adult female rabbits were used for the 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 of 0.5ml test item (as such, undiluted) 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, no erythema and oedema was observed at 1 hour of observation. At 24, 48 and 72 hours observation no erythema and no oedema was observed.Hence the 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 wereobservedfor erythema and oedemaat 1, 24, 48 and 72 hours after patch removal, evaluatedand graded as per draize method.

In Animal No. 2 & 3, at 1, 24, 48 and 72 hours, animal nos. 2 and 3 showed no erythema and no oedema.

The other untreated side revealed no erythema and no oedema and was found to be normal throughout the experimental 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.

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 and being classified as “Not

Classified” as per CLP Classification.

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

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
data is from experimental reports
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 the 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
- Sex: male
- Source:Procured from LIVEON BIOLABS PVT. LTD., Karnataka, India
- Age at study initiation:3.0 to 4.5 Months (Approximately)
- Weight at study initiation: Minimum: 1.568 kg and Maximum: 1.742 kg (Prior to Treatment)
- Health Status :Healthy young adult animals 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 6 days (Animal No.-1) and 9 days (Animal No. 2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 19.00 °C and Maximum: 20.80 °C
- Humidity (%):Minimum: 45.10 % and Maximum: 65.80 %
- 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 and 72 hours after instillation of test item.
Duration of post- treatment incubation (in vitro):
no data available
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 fluorescent strips.
Irritation parameter:
cornea opacity score
Basis:
animal: #1,#2 and #3
Time point:
other: After 1, 24, 48 and 72 Hours
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: After 1, 24, 48 and 72 Hours
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: After 1, 24, 48 and 72 Hours
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: After 1, 24, 48 and 72 Hours
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 1, 24, 48 and 72 Hours
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
iris score
Basis:
animal: #1,#2 and #3
Time point:
other: After 1, 24, 48 and 72 Hours
Score:
0
Max. score:
2
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: After 1 and 24 Hours
Score:
1
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
other: After 1 and 24 Hours
Score:
1
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: After 1 and 24 Hours
Score:
1
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
conjunctivae score
Basis:
animal: #1,#2 and #3
Time point:
other: After 48 and 72 Hour
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: After 1 Hours
Score:
1
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: After 1 Hours
Score:
1
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: After 1 Hours
Score:
1
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
chemosis score
Basis:
animal: #1,#2 and #3
Time point:
other: After 1, 24, 48 and 72 Hours
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritant / corrosive response data:
Untreated eye of all the three rabbits was normal throughout the experimental period of 72 hours.

The following grading scores were observed in treated eye of tested rabbits.
Observation at 1 hour after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was 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 - Some blood vessels definitely hyperaemic (injected) was observed 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% damage in all the animals.

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 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 3 at 24, 48, 72 hours (at treated site) for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively.
Other effects:
Clinical Observation
No systemic toxicity was observed in treated rabbits during the experimental period

Mortality
No mortality was observed during the observation period.

Body weight
Increase in body weights in all the treated animals at termination when compared to day 0 was observed

Table 1 : Individual Animal Eye Irritation Scores

 

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

Animal numbers

1

2

3

Application site

Right

Right

Right

Eye reactions

At hours

At hours

At hours

*

1

24

48

72

*

1

24

48

72

*

1

24

48

72

Corneal opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Area of opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Iris

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctiva

0

1

1

0

0

0

1

1

0

0

0

1

1

0

0

Chemosis

0

1

0

0

0

0

1

0

0

0

0

1

0

0

0

Corneal damage %

0

./.

0

0

./.

0

0

./.

0

In Treated area Dose:Untreated                                                                             Sex:Male

Animal Numbers

1

2

3

Application Side

Left

Left

Left

Eye Reactions

At hour

At hour

At hour

*

1

24

48

72

*

1

 24

48

72

*

1

24

48

72

Corneal Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Area of Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Iris

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctiva

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Chemosis

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Corneal Damage%

0

./.

0

0

./.

0

0

./.

0

Key:*= Pre-exposure eye examination.

      ./.= Not Applicable

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

0.33

0.33

0.33

Chemosis

0.00

0.00

0.00

 

 

Eye Irritation Scores - Mean Values after 24, 48, 72 Hours (Untreated 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 reaction at 24, 48 and 72 hours

Number of the Observations (3)

Table 2 : Individual AnimalClinicalSigns

 

Sex:Male

Animal No.

Days (Post application observation)

0

1

2

3

1

1

1

1

1

2

1

1

1

1

3

1

1

1

1

Key:1 = Normal

Table 3: Individual Animal Body Weight

Sex :Male

Animal No.

Animal Body Weight (kg)

Prior to application

At termination

1

1.568

1.608

2

1.590

1.674

3

1.742

1.818

Key:kg = Kilogram

Interpretation of results:
other: not irritating
Conclusions:
The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours (at treated site)for corneal opacity, iris, conjunctiva and chemosis were determined to be 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively. Hence, it was concluded that the test chemical was Non-Irritating to the eyes of Male New Zealand White rabbits under the experimental conditions tested and is Classified as “Category- Not Classified" as per CLP criteria for Classification.
Executive summary:

An Acute Eye Irritation/Corrosion Study for the test substance was performed in Rabbit as per OECD guideline No. 405. 3 male New Zealand White rabbits were used for the study.

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 as such test itemwas instilled in the other (treated) eye of each rabbit. The eye was observed at 1, 24, 48 and 72 hour after test item instillation.Ophthalmoscope was used for scoring of eye lesions. In the initial test,0.1 ml of as such test itemwas instilled 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 hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3);0.1 ml of as such test itemwas 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 was normal throughout the experimental period of 72 hours. The following grading scores were observed in treated eye of tested rabbits.

Observation at 1 hour after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity-Zero in all the animals;Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was 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 - Some blood vessels definitely hyperaemic (injected) was observed 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% damage in all the animals.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 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 3 at 24, 48, 72 hours (at treated site)for corneal opacity, iris, conjunctiva and chemosis were determined to be 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively.

Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed within 72 hours. 

Hence, it was concluded that the test chemical was Non-Irritating to the eyes of Male New Zealand White rabbits under the experimental conditions tested and is Classified as “Category- Not Classified" as per CLP criteria for Classification.

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 to cause dermal irritation in living organisms to a greater or lesser extent. The studies are based on in vivo experiments in rabbits, guinea pigs, humans as well as estimated data for the test chemical. The results are summarized as follows:

Acute Dermal Irritation/corrosion Study of the test chemical was performed on Rabbits as per OECD Guideline No. 404.

Three healthy young adult female rabbits were used for the 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 of 0.5ml test item (as such, undiluted) 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 oedema was observed at 1 hour of observation. At 24, 48 and 72 hours observation no erythema and no oedema was observed.Hence the 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 after patch removal, evaluated and graded as per Draize method.

In Animal No. 2 & 3, at 1, 24, 48 and 72 hours, animal nos. 2 and 3 showed no erythema and no oedema.

The other untreated side revealed no erythema and no oedema and was found to be normal throughout the experimental 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.

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 and being classified as “Not Classified” as per CLP Classification.

This is supported by the results of a skin irritation study performed in humans to assess the irritation potential of the test chemical. The test chemical was tested 5% in petrolatum on human volunteers in a 48 hours closed patch test.

The test chemical was not irritating to humans after 48 hours exposure.

These results are supported by a skin irritation study performed in rabbits to assess the irritation potential of the test chemical. 500 mg undiluted test chemical was applied on the intact and abraded skin of rabbits under occlusion for 24 hours.

Slight irritation was observed when rabbits were tested with undiluted test chemical for 24 hours. Hence, the test chemical can be considered to be irritating to skin.

Skin irritation effects were also estimated by four different models i.e, Battery, leadscope, SciQSAR and CASE Ultra used within Danish QSAR database for test chemical.

Based on estimation, No severe skin irritation effect were known when test chemical was exposed to rabbit skin.

Hence, the test chemical can be considered not irritating to skin.

The estimated result is further supported by a preliminary irritation test performed on guinea pigs to determine the maximum non-irritating concentration for the Open Epicutaneous test. 6-8 male/female guinea pigs were used for the study.

0.025 ml of each test concentration (e.g, 100, 30, 10 and 3%) inethanol, acetone, H20, petroleum, PEG and/or other suitable vehicles was applied to the one of the areas measuring 2 cm2 of the flank skin previously clipped and marked with a circular stamp. Reactions are read 24 h after the application of the test material. The minimal irritating and the maximal nonirritating concentrations were determined by an all-or-none criterion. The minimal irritating concentration is defined as the lowest one causing skin irritation. The maximal nonirritating concentration is defined as the highest one not causing macroscopic skin reactions in any of the animals.

The test chemical was considered to be not irritating to guinea pig skin.

The above results are supported by an acute Dermal Irritation/corrosion Study was performed in Rabbits, as per OECD guideline No. 404 to evaluate the irritation potential of test chemical. Three healthy young adult female rabbits were used for conducting acute dermal irritation/corrosion 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 of 0.5 ml test item was applied to the skin, over an area of approximately 6 x 6 cm clipped of hair on one side of rabbits. The other untreated side was kept as control area and 0.5 ml of distilled water was applied at this site. At the end of 4 hours, the gauze patch was removed and test item application site was wiped with water without altering the integrity of the epidermis.  Initially, the test item was applied to the clipped area of skin of one rabbit. The test site was covered with gauze patch. After 4 hours of exposure in animal no.1, no erythema and oedema was observed at 1 hour of observation. At 24, 48 and 72 hours observation no erythema and no oedema was observed. Hence the 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 after patch removal, evaluated and graded as per Draize method. The individual mean score at 24, 48 and 72 hours for treated site for animal nos. 1, 2 and 3 were 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively. No erythema and no oedema (skin irritation) were observed at the end of 72 hour observation period after patch removal. 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.

Even though the results of rabbit study substantiate the claim that undiluted test chemical can indeed be irritating to skin, but the results from the remaining in vivo as well as estimation study present a stronger evidence of its not irritating properties.

Hence, taking all these factors into consideration, the test chemical can be considered to be not irritating to skin. Comparing the above annotations with the criteria of CLP Regulation, the 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 in living organisms to a greater or lesser extent. The studies are based on in vivo experiments in rabbits for the test chemicals. The results are summarized as follows:

An Acute Eye Irritation/Corrosion Study for the test substance was performed in Rabbit as per OECD guideline No. 405. 3 male New Zealand White rabbits were used for the study.

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 as such test itemwas instilled in the other (treated) eye of each rabbit. The eye was observed at 1, 24, 48 and 72 hour after test item instillation.Ophthalmoscope was used for scoring of eye lesions. In the initial test,0.1 ml of as such test itemwas instilled 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 hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3);0.1 ml of as such test item 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 was normal throughout the experimental period of 72 hours. The following grading scores were observed in treated eye of tested rabbits.

Observation at 1 hour after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity-Zero in all the animals;Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was 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 - Some blood vessels definitely hyperaemic (injected) was observed 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% damage in all the animals. 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 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 3 at 24, 48, 72 hours (at treated site)for corneal opacity, iris, conjunctiva and chemosis were determined to be 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively.

Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed within 72 hours. 

Hence, it was concluded that the test chemical was Non-Irritating to the eyes of Male New Zealand White rabbits under the experimental conditions tested and is Classified as “Category- Not Classified" as per CLP criteria for Classification.

This is supported by the results of a similar OECD guideline no. 405 was performed in Rabbits to evaluate the eye irritation potential of the test chemical. 3 male New Zealand White rabbits were used for the study.

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, 72 hours after test item instillation. Ophthalmoscope was used for scoring of eye lesions.In the initial test,0.1 ml of as such test item was instilled 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 hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3);0.1 ml of as such test item 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 was normal throughout the experimental period of 72 hours.

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

Observation at 1 hour after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity-Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was 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 - Some blood vessels definitely hyperaemic (injected) was observed 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% damage in all the animals. 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 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 3 at 24, 48, 72 hours (at treated site) for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively.

Under the experimental conditions tested, the test chemical was observed to be Non Irritant to New Zealand White male rabbit eyes.

The results from the Guideline studies indicate a very strong possibility that the test chemical cannot cause any irritation to eyes.The test chemical can be considered to be not irritating to eyes. Comparing the above annotations with the criteria of CLP Regulation, the test chemical can be classified under the category “Not Classified”.

Justification for classification or non-classification

Based on the available resutls the test chemical can be considered to lack the potential to cause irritation/corrosion to eyes and skin. Thus the test chemical can be considered to be not irritating to skin and eyes. Hence, the test chemical can be classified under the category "Not Classified" as per CLP Regulation.