Phenethyl phenylacetate

Endpoint summary

Description of key information

Skin Irritation    

Acute Dermal Irritation/corrosion Study of the test chemical in Rabbits, was performed as per OECD guideline No. 404. Three healthy young adult New Zealand White female rabbits were used for conducting acute dermal irritation/corrosion study.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. 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 and can be Classified under the category "Not Classified” as per CLP criteria for Classification.

Eye Irritation

Acute Eye Irritation/Corrosion Study of the test chemical in Rabbits, was performed 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 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 considered to be Non Irritant to New Zealand White male rabbit eyes and was thus Classified as "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

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 or test system and environmental conditions:

TEST ANIMALS
- Source: Procured from GENTOX BIOSERVICES PVT. LTD.
- Age at study initiation: 4.0 to 5.5 Months (Approximately)
- Weight (Prior to Treatment): Minimum: 1.990 kg & Maximum: 2.186 kg
- 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, 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 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:

unchanged (no vehicle)

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

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 #2

Time point:

other: At 1 hours

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 24 hrs

Remarks on result:

other: Very slight erythema (barely perceptible)

Irritation parameter:

erythema score

Basis:

animal: #1 ,#2 and #3

Time point:

24/48/72 h

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:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The following were observed in treated rabbits.
The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method.

Animal No. 1, showed no erythema and oedema at 1, 24, 48 and 72 hours of observation. In animal no. 2, at 1 hour observation post patch removal, revealed very slight erythema (barely perceptible) and no oedema whereas animal no. 3 showed no erythema and oedema. At 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 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

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 was observed

Table 1: Skin Reaction

In Treated area Dose: 0.5 ml of test item (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	1	0	0	0	0	0	0	0
3		Right	0	0	0	0	0	0	0	0

In Control area Dose: 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		Left	0	0	0	0	0	0	0	0

Key: h = Hour.

Erythema                                                                                                       Oedema

0 =No erythema                                                                                           0 =No oedema

1 = Very slight erythema(barely perceptible)

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 Body Weight

Sex: Female

Animal No.	Body Weight (kilogram)
	Prior to Dosing	At termination
1	2.136	2.178
2	1.990	2.080
3	2.186	2.240

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:

No erythema and no oedema (skin irritation) were found at the end of 72 hour observation period after patch removal.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. 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 and can be Classified under the category "Not Classified” as per CLP criteria for Classification.

Executive summary:

Acute Dermal Irritation/corrosion Study of the test chemical in Rabbits, was performed as per OECD guideline No. 404. Three healthy young adult New Zealand White 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 chemical 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 chemical application site was wiped with water without altering the integrity of the epidermis.Initially, the test chemical 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 chemical. The patch was removed after 4 hours and rabbits were observed for erythema and oedemaat 1, 24, 48 and 72 hours after patch removal, evaluatedand graded as per Draize method. In animal no. 2, at 1 hour observation post patch removal, revealed very slight erythema (barely perceptible) and no oedema whereas animal no. 3 showed no erythema and oedema. At 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 observed to be normal throughout the experimental period. No erythema and no oedema (skin irritation) were observed at the end of 72 hour observation period after patch removal.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. 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 and can be Classified under the category "Not Classified” as per CLP criteria for 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

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.
- Age at study initiation: 3.0 to 4.5 Months (Approximately)
- Weight at study initiation: Minimum: 1.822 kg and Maximum: 2.146 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, 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 treated 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:

7 d

Score:

0

Max. score:

3

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:

7 d

Score:

0

Max. score:

3

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:

7 d

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:

7 d

Score:

0

Max. score:

3

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:

24/48/72 h

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:

24/48/72 h

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,#2 and #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

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:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

not specified

Remarks on result:

no indication of irritation

Remarks:

For treated eyes

Irritant / corrosive response data:

The following were observed in treated rabbits.
Observation at 1 hour after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) 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 experimental 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 Side	Right					Right					Right
Eye Reactions	At hour					At hour					At hour
	*	1	24	48	72	*	1	24	48	72	*	1	24	48	72
Corneal Opacity	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Area of Opacity	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Iris	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Conjunctiva	0	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	Right					Right					Right
Eye Reactions	At hour					At hour					At hour
	*	1	24	48	72	*	1	24	48	72	*	1	24	48	72
Corneal Opacity	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Area of Opacity	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Iris	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Conjunctiva	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Chemosis	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Corneal Damage%	0	./.	0			0	./.	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 Animal Clinical Signs

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

Animal No.	Animal Body Weight (kg)
	Prior to application	At termination
1	2.146	2.180
2	2.146	2.198
3	1.822	1.886

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 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 considered to be Non Irritant to New Zealand White male rabbit eyes and is thus Classified as “Category- Not Classified as Eye Irritant” as per CLP criteria for Classification.

Executive summary:

Acute Eye Irritation/Corrosion Study of the test chemical in Rabbits, was performed 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 test chemical (as such) was instilled in the other (treated) eye of rabbits. The eye was observed at 1, 24, 48, 72 hours after test chemical instillation. Ophthalmoscope was used for scoring of eye lesions. In the initial test,0.1 ml of as such test chemical 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 chemical 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 chemical 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 chemical 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 chemical 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 considered to be Non Irritant to New Zealand White male rabbit eyes and was thus Classified as "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               

Various studies have been reviewed to evaluate the dermal irritation/corrosion potential of the test chemical. These include in vivo experimental studies performed on rabbits as well as humans for the test chemical. The results are mentioned below:

Acute Dermal Irritation/corrosion Study of the test chemical in Rabbits, was performed as per OECD guideline No. 404. Three healthy young adult New Zealand White 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 chemical 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 chemical application site was wiped with water without altering the integrity of the epidermis. Initially, the test chemical 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 were observed at 1 hour of observation. At 24, 48- and 72-hours observation no erythema and no oedema were observed. Hence the confirmatory test was conducted on additional two rabbits (No. 2 and 3) to confirm the non-irritant nature of the test chemical. 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, at 1-hour observation post patch removal, revealed very slight erythema (barely perceptible) and no oedema whereas animal no. 3 showed no erythema and oedema. At 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 observed to be normal throughout the experimental period. No erythema and no oedema (skin irritation) were observed at the end of 72-hour observation period after patch removal. 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. 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 and can be Classified under the category "Not Classified” as per CLP criteria for Classification.

This result is supported by similar OECD 404 Guideline study performed to assess the irritation parameter of the test chemical.3 female New Zealand White rabbits were used for the study. The rabbits were prepared 24 hrs prior to application of test product. The furs from the dorsal area of trunk of animals were removed with electric clippers exposing an area measuring approximately 6 cm2 of body surface area of animal. The care was taken such that abrasion penetrated the Stratum corneum only and not dermis. 0.5 gm of the test compound was applied on a small area 6cm2 of the intact test site. Each site of application was covered with an impervious dressing which was secured with an adhesive tape. The animals were housed individually and restrained by the use of plastic collars. After patch removal [4 hours later] the unabsorbed test chemical was removed and the test site was washed with lukewarm water. The intact skin site of application was observed for erythema and edema at 1, 24,48 and 72 hours till 14 days after application and scored according to Draize method. The test chemical did not produce erythema or edema throughout the observation period of 14 days. The dermal irritation index was calculated to be 0.00. The test chemical did not produce any clinical signs of toxicity throughout the study. Based on the score and observations, the test chemical can be considered not irritating to skin and can be classified under the category 'Not Classified' as per CLP Regulation.

The results from the above Guideline studies are supported by a study performed on humans. 2% test chemical in petrolatum was applied under occlusive conditions for 48 hours to the skin of human volunteers. The test sites were observed for signs of irritation. The test chemical failed to produce any signs of irritation after 48 hours closed patch test. Hence, it can be considered to be not irritating to skin.

These results are further supported by a repeated insult patch test performed to evaluate the dermal irritation potential of the test chemical. 2% test chemical was applied to the skin of humans and observed for effects. No signs of dermal irritation were observed when human volunteers were exposed to the test chemical in a repeated insult patch procedure. Hence, the test chemical was considered to be not irritating to skin.

Available results indicate that the test chemical indeed lacks the potential to cause skin irritation in living organisms. Hence, the test chemical can be considered to be not irritating to skin and can be classified under the category "Not Classified" as per CLP Regulation.

Eye Irritation

Various studies have been reviewed to evaluate the eye irritation/corrosion potential of the test chemical. These include in vivo experimental studies performed on the test chemical. The results are as follows:

Acute Eye Irritation/Corrosion Study of the test chemical in Rabbits, was performed 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 test chemical (as such) was instilled in the other (treated) eye of rabbits. The eye was observed at 1, 24, 48, 72 hours after test chemical instillation. Ophthalmoscope was used for scoring of eye lesions. In the initial test,0.1 ml of as such test chemical 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 chemical 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 chemical 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 chemical 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 chemical 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 considered to be Non Irritant to New Zealand White male rabbit eyes and was thus Classified as "Not Classified ”as per CLP criteria for Classification.

This result is supported by a similar OECD 405 Guideline study performed on female New Zealand White rabbits to assess the irritation parameter of the test chemical. 3 female New Zealand White rabbits were used for the study. 0.1 ml of the test chemical was moistened with water was instilled in the conjunctival sac of rabbits after gently pulling the lower lid away from the eyeball. The other eye which remained untreated served as a control. The ocular lesions were evaluated at 1, 24, 48 and 72 hours after the treatment. The grades of ocular reactions (conjunctiva, cornea and iris) were recorded at each observation. To determine the reversibility of the effect the animals were observed normally for 21 days. Any other lesions in the eye viz pannus, staining were observed and scored accordingly. Examination of reactions was facilitated by use of biomicroscope and hand slit lamp. Individual animal weights before and during the study was observed. The overall irritation index of the test chemical was 0.0 after 72 hours. Also, the test chemical did not produce any clinical signs of toxicity throughout the examination period of 21 days. Hence, under the test conditions, the test chemical can be concluded to be not irritating to New Zealand White rabbit eyes. Based on these observations and scores, the test chemical can be classified under the category "Not Classified" as per CLP Regulation.

Available results from the Guideline studies indicate that the test chemical indeed lacks the potential to cause eye irritation/damage to test animals. Hence, the test chemical can be considered to be not irritating to eyes and can be classified under the category "Not Classified" as per CLP Regulation.

Justification for classification or non-classification

Available results indicate that the test chemical indeed lacks the potential to cause irritation/damage to skin and eyes. Hence, the test chemical can be considered to be not irritating to skin and eyes and can be classified under the category "Not Classified" as per CLP Regulation.