Registration Dossier

Skin Irritation:

The Primary Irritation Index (PII) for the test chemical after 14 days of observation was 0.0. Also the test chemical did not produce any clinical signs of toxicity throughout the examination period of 14 days.

Hence, under the test conditions, the test chemical can be concluded to be not irritating to New Zealand White rabbit skin.

Eye Irritation:

The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.00, 0.00; 0.00, 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, 0.00, respectively. Hence under the experimental test conditions, the test chemical can be considered as “Non Irritant” to New Zealand White female rabbit eyes.

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

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:

To assess the dermal irritation parameter of the test chemical in accordance with OECD 404

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Sex: Female
- Age at study initiation: 10-12 weeks
- Weight at study initiation: 2 kg+ 200g
- Housing:animals were housed individually in stainless steel cages provided with steel mesh bottom and facilities for food and water
- Diet (e.g. ad libitum): Pelleted feed supplied by Pranav Agro
- Water (e.g. ad libitum):Community tap water kept in glass bottles, ad libitum
- Acclimation period: The healthy rabbits selected for study were acclimatized to laboratory conditions one week prior to testing
- Identification: By cage tag and corresponding color body marking

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-25 degC
- Humidity (%): relative humidity - 40-60%
- Air changes (per hr): Air conditioned rooms with 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

0.5gm

Duration of treatment / exposure:

4 hours

Observation period:

24,48 and 72 hours after patch removal till 14days

Number of animals:

3- [1- initial test; 2- confirmatory test]

Details on study design:

TEST SITE
- Area of exposure: 6 sq.cm of the dorsal area of the trunk
- % coverage: 6 sq.cm
- Type of wrap if used: impervious dressing with an adhesive tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: the patches were removed and the site of application was cleaned with lukewarm water after patch removal

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : The treated sites were examined and scored at 60 minutes, 24,48 and 72 hours after application.

SCORING SYSTEM:
- Method of calculation: Draize method

Irritation parameter:

erythema score

Basis:

mean

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:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The test compound didnot produce any signs of irritation through out the observation period

Other effects:

The test compound did not produce any clinical signs of toxicity through out the study.

TABLE - 1

INDIVIDUAL ANIMAL DERMAL IRRITATION SCORES

Rabbit No.	Sex	INTACT SKIN
		3 Min.		4 Hours		24 Hours		48 Hours		72 Hours		14 days
		Erythema	Oedema	Erythema	Oedema	Erythema	Oedema	Erythema	Oedema	Erythema	Oedema	Erythema	Oedema
01	F	0	0	0	0	0	0	0	0	0	0	0	0
02	F	-	-	0	0	0	0	0	0	0	0	0	0
03	F	-	-	0	0	0	0	0	0	0	0	0	0
Total		0	0	0	0	0	0	0	0	0	0	0	0
Mean		0	0	0	0	0	0	0	0	0	0	0	0
Grand Total		0.00

Dermal Irritation Index: 0.0/4 = 0.0

CLINICAL SIGNS

SEX	ANIMAL NO.	Time (Min.)	Time (Hours)					Time (Day)
SEX	ANIMAL NO.	3	1	4	24	48	72	14
FEMALE	01	N	N	N	N	N	N	N
	02	N	N	N	N	N	N	N
	03	N	N	N	N	N	N	N

N: No Clinical Signs

C: Clinical Signs Observed

Interpretation of results:

other: not irritating

Conclusions:

The Primary Irritation Index (PII) for the test chemical after 14 days of observation was 0.0. Also the test chemical did not produce any clinical signs of toxicity throughout the examination period of 14 days.
Hence, under the test conditions,the test chemical can be concluded to be not irritating to New Zealand White rabbit skin.

Executive summary:

A dermal irritation study was conducted on New Zealand white rabbits in accordance with OECD 404 to assess the irritation parameter of the test chemical. 3 female New Zealand White rabbits were used for the study. The animals 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 6 square cms 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 Primary Irritation Index (PII) for the test chemical after 14 days of observation was 0.0. Also the test chemical did not produce any clinical signs of toxicity throughout the examination period of 14 days.

Hence, under the test conditions,the test chemical can be concluded to be not irritating to New Zealand White rabbit skin.

Endpoint conclusion:

no adverse effect observed (not irritating)

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

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:

To assess the eye irritation potential of the test chemical according OECD 405 Guidelines

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Sex: Female
- Age at study initiation: 10-12 weeks
- Weight at study initiation: 2 kg+ 200g
- Housing:animals were housed individually in stainless steel cages provided with steel mesh bottom and facilities for food and water
- Diet (e.g. ad libitum): Pelleted feed supplied by Pranav Agro
- Water (e.g. ad libitum):Community tap water kept in glass bottles, ad libitum
- Acclimation period: The healthy rabbits selected for study were acclimatized to laboratory conditions one week prior to testing
- Identification: By cage tag and corresponding color body marking

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-25 degC
- Humidity (%): relative humidity - 40-60%
- Air changes (per hr): Air conditioned rooms with 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 gm

Duration of treatment / exposure:

single exposure

Observation period (in vivo):

1,24,48 and 72 hours after test substance application

Duration of post- treatment incubation (in vitro):

no data available

Number of animals or in vitro replicates:

3- [Initial test =1; and confirmatory tes = 2]

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: The eyes of the test animal were washed

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein: The examination of the reactions were facilatated by use of biomicroscope and hand slit lamp.

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

110

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

0.1gm of the test chemical when applied to the eyes of New Zealand White rabbits did not produce any lesions such as pannus, staining through out the observation period of 72 hours.

Other effects:

There were no other signs observed through out the observation period of 21 days.

TABLE- 1 GRADING OF OCULAR LESIONS

S.NO/ SEX		OBSERVATION	Score				Total	Total Score
1/F		OBSERVATION	1 hr	24hrs	48 hrs	72 hrs	Total	Total Score
	Cornea	A. Opacity-Degree of Density	0	0	0	0	0	0×0×5=0
	Cornea	B. Area of Cornea Involved	0	0	0	0	0	0×0×5=0
	Iris	A. Values	0	0	0	0	0	0×5=0
	Conjunctivae	A. Redness	1	0	0	0	0	0+0 +0×5=0
		B. Chemosis	0	0	0	0	0
		C. Discharge	0	0	0	0	0
2/F	Cornea	A. Opacity-Degree of Density	0	0	0	0	0	0×0×5=0
	Cornea	B. Area of Cornea Involved	0	0	0	0	0	0×0×5=0
	Iris	A. Values	0	0	0	0	0	0×5=0.0
	Conjunctivae	A. Redness	1	0	0	0	0	0+0 +0×5=0
		B. Chemosis	0	0	0	0	0
		C. Discharge	0	0	0	0	0
3/F	Cornea	A. Opacity-Degree of Density	0	0	0	0	0	0×0×5=0.0
	Cornea	B. Area of Cornea Involved	0	0	0	0	0	0×0×5=0.0
	Iris	A. Values	0	0	0	0	0	0×5=0.0
	Conjunctivae	A. Redness	1	0	0	0	0	0+0 +0×5=0
		B. Chemosis	0	0	0	0	0
		C. Discharge	1	1	0	0	2
Grand total								0
Mean								0
Eye Irritation Scoring Index								0

Interpretation of results:

other: not irritating

Conclusions:

0.1gm of the test chemical when applied to the eyes of New Zealand White rabbits did not produce any lesions such as pannus, staining through out the observation period of 72 hours. The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.00, 0.00; 0.00, 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, 0.00, respectively. Hence under the experimental test conditions, the test chemical can be considered as “Non Irritant” to New Zealand White female rabbit eyes.

Executive summary:

Acute Eye Irritation/Corrosion Study of test chemical in Rabbits was performed as per OECD guideline no. 405.

3 female New Zealand White Rabbits free from injury of eye were selected for the study. The eyes of all the rabbits were examined 24 hours prior to treatment.

The test compound in the amount of 0.1 gm was applied as such and then placed in the conjunctival sac of one eye of each animal after gentle pulling the lower eye lid away from the eye ball. The eye lids were then gently held together for about one second to prevent the loss of the material. The other eyes which remains untreated, served as control. The eyes of the test animal were not washed for at least 24 hours following instillation of the test compound. After 24 hours, eyes were washed gently with the luke warm water. The eye was observed at 1, 24, 48 and 72 hours after test item instillation.

In the initial test, 0.1 ml of test item was applied into the conjunctival sac of the right eye of animal no.1 whereas the left eye of the rabbit served as the control. As animal no. 1 showed no severe ocular lesions till 24 hour observation hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 0.1 ml of test item was instilled into the conjunctival sac of right eye of both the rabbits and left eye served as the control. There were no other signs observed through out the observation period of 21 days.

The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.00, 0.00; 0.00, 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, 0.00, respectively. Hence under the experimental test conditions, the test chemical can be considered as “Non Irritant” to New Zealand White female rabbit eyes.

Endpoint conclusion:

no adverse effect observed (not irritating)

Endpoint conclusion:

no study available

Skin Irritation

A dermal irritation study was conducted on New Zealand white rabbits in accordance with OECD 404 to assess the irritation parameter of the test chemical. 3 female New Zealand White rabbits were used for the study. The animals 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 6 square cms 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 Primary Irritation Index (PII) for the test chemical after 14 days of observation was 0.0. Also the test chemical did not produce any clinical signs of toxicity throughout the examination period of 14 days.

Hence, under the test conditions, the test chemical can be concluded to be not irritating to New Zealand White rabbit skin.

Eye Irritation

Acute Eye Irritation/Corrosion Study of test chemical in Rabbits was performed as per OECD guideline no. 405.

3 female New Zealand White Rabbits free from injury of eye were selected for the study. The eyes of all the rabbits were examined 24 hours prior to treatment.

The test compound in the amount of 0.1 gm was applied as such and then placed in the conjunctival sac of one eye of each animal after gentle pulling the lower eye lid away from the eye ball. The eye lids were then gently held together for about one second to prevent the loss of the material. The other eyes which remains untreated, served as control. The eyes of the test animal were not washed for at least 24 hours following instillation of the test compound. After 24 hours, eyes were washed gently with the luke warm water. The eye was observed at 1, 24, 48 and 72 hours after test item instillation.

In the initial test, 0.1 ml of test item was applied into the conjunctival sac of the right eye of animal no.1 whereas the left eye of the rabbit served as the control. As animal no. 1 showed no severe ocular lesions till 24 hour observation hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 0.1 ml of test item was instilled into the conjunctival sac of right eye of both the rabbits and left eye served as the control. There were no other signs observed through out the observation period of 21 days.

The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.00, 0.00; 0.00, 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, 0.00, respectively. Hence under the experimental test conditions, the test chemical can be considered as “Non Irritant” to New Zealand White female rabbit eyes.

Available results from the Guideline studies indicate a very strong possibility that the test chemical lacks the potential to cause any irritation to eyes and skin. Hence, the test chemical can be considered to be not irritating to eyes and skin. It can be further classified under the category “Not Classified” as per CLP regulation.