p,p',p''-tris(dimethylamino)trityl alcohol

Endpoint summary

Description of key information

Skin irritation:

The dermal irritation potential of target chemical was assessed in various in-vivo experimental studies.Based on the available studies,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 various in-vivo experimental studies.Based on the available studies,it can be concluded that the test chemical is unable to cause eye 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''.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data of test substances

Justification for type of information:

Data for the target chemical is summarized based on the available studies

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

WoE report is based on skin irritation studies as- WoE-2 and WoE-3.
The eye irritation studies were condcuted on rabbits to assess its skin irritating effects.

GLP compliance:

not specified

Species:

other: 2.rabbit 3.rat

Strain:

other: 2.Russian breed 3.Sprague-Dawley

Details on test animals or test system and environmental conditions:

2.TEST ANIMALS
- Source: Six rabbits (3 males/3 females) of the Russian breed
- Weight at study initiation: 1.5 to 2 kgs
- Housing: V2A wire cages
- Diet (e.g. ad libitum): food (NAFAG, Gossau SG, rabbit food)
- Water (e.g. ad libitum): water ad libitum

3.
TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Females (if applicable) nulliparous and non-pregnant: No data available
- Age at study initiation: Young adult male and female rats aged between 8 – 12 weeks were used.
- Weight at study initiation: The weight range of approximately 218.6 to 246.8 grams at initiation of dosing.
Body weights at the start :
Male
Mean : 244.22 g (= 100 %)
Minimum : 241.7 g (- 1.03 %)
Maximum : 246.8 g (+ 1.06 %)
Total No. of animals : 5
Female
Mean : 221.38 g (= 100 %)
Minimum : 218.6 g (- 1.26 %)
Maximum : 225.6 g (+ 1.91 %)
Total No. of animals : 5
- Identification: Each rat was individually identified by the cage number.
- Fasting period before study: No data available
- Housing: The rats were individually housed in polycarbonate cages with paddy husk as bedding.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.7 to 21.8 degree centigrade.
- Humidity (%): 56.2% to 60.1%
- Air changes (per hr): Ten to fifteen air changes per hour.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: 13-07-2017 to 28-07-2017

Type of coverage:

other: 2.occlusive 3.semiocclusive

Preparation of test site:

other: 2.shaved 3.clipped

Vehicle:

other: 2.unchanged (no vehicle) 3.unchanged (no vehicle)

Controls:

no

not specified

Amount / concentration applied:

2.
0.5 g (500mg)

3.
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Concentration (if solution): No data available

VEHICLE
- Amount(s) applied (volume or weight with unit): No data available
- Concentration (if solution): No data available
- Lot/batch no. (if required): No data available
- Purity: No data

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): No data available
- Concentration (if solution): No data available

POSITIVE CONTROL
- Amount(s) applied (volume or weight): No data available
- Concentration (if solution): No data available


3.

Duration of treatment / exposure:

2.
24 hours

3.
24 hours

Observation period:

2.
72 hours

3.
14 days

Number of animals:

2.
6 (3 males/3 females)

3.
10 (5/sex).

Details on study design:

2.
Six rabbits (3 males/3 females) of the Russian breed,weighing 1.5 to 2 kgs, were shaved on the whole back and flanks two days before treatment with an electric clipper. In order to prevent injuries by bites the rabbits were kept separately in V2A wire cages, where they had access to food (NAFAG, Gossau SG, rabbit food) and water ad libitum. The shaven skin on the left side was slightly scarified immediately before treatment.A gauze patch 2.5 x 2.5 cm was soaked with the test substance and immediately applied to the prepared skin. The patch was covered with a plastic film 5 x 5 cm, which was fixed, to the body with adhesive tape.
TK 11678 was applied to each side in quantities of 0.5 g. Before application a 50 % polyethylene glycol trituration (PEG 400) was made. The gauze patches were removed 24 hours after the application. The reaction of the skin was appraised upon removal and 72 hours after it.

3.TEST SITE
- Area of exposure: Trunk (dorsal surface and sides from scapular to pelvic area)
- % coverage: Approximately 10% of the body surface area.
- Type of wrap if used: Porous gauze dressing and non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Distilled water was used to remove residual test item.

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : Dermal reaction was observed daily for study period of 14 days.

SCORING SYSTEM: Draize Method.

OTHER OBSERVATIONS
Type and Frequency of Tests, Analyses and Measurements

Viability:Twice daily.

Clinical Observations and General Appearance:
Animals were observed for clinical signs, mortality, until sacrifice.
Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time.
The observations were included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behavior pattern.

Body weights:
Individual animal body weights were recorded pre-test (prior to administration of the test item), day 7 and at termination on day 14.

Gross Pathology:
Necropsy was performed on animals surviving at the end of the study. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique (day 15).

Histopathology:
No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.

Irritation parameter:

primary dermal irritation index (PDII)

Remarks:

2.

Basis:

mean

Time point:

other: 24 and 74 hours

Score:

0

Max. score:

8

Remarks on result:

other: Not Irritant

Irritation parameter:

erythema score

Remarks:

3.

Basis:

mean

Time point:

14 d

Score:

0

Max. score:

4

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

3.

Basis:

mean

Time point:

14 d

Score:

0

Max. score:

4

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

3.Overall result:
Sex : Male
Group I -
Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.

Sex : Female
Group I -
Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.

Other effects:

3.Other effects:
Clinical Signs of Toxicity and Mortality
Sex : Male
Group I -
Animal treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. All animals survived through the study period of 14 days.

Sex : Female
Group I -
Animal treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. All animals survived through the study period of 14 days.

Body Weight
Sex : Male
Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 19.64% and 29.31% respectively.

Sex : Female
Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 11.09% and 15.97% respectively.

Gross Pathological Findings
Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group.

2.

The primary irritation index as the measure of the acute skin irritation provoked by the substance is defined as the average of all the ratings found after 24 and 72 hours on the intact and slightly scarified skin. Compounds producing combined averages (primary irritation indexes) of 2 or less are only mildly irritating; whereas those with indexes from 2 to 5 are moderate irritants, and those with scores above 6 are considered severe irritants.

3.

Individual Animal - Evaluation of Dermal Reaction

 

Laboratory Test Item Code :TAS/122/042

Test System : Sprague Dawley Rat

Sex : Male  

Group : I

Dose  : 2000 mg/kg body weight

Animal	Dermal	D A Y S
No.	Reaction	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14
1	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
3	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
4	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
5	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

Sex : Female  

Group : I

Dose  : 2000 mg/kg body weight

Animal	Dermal	D A Y S
No.	Reaction	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14
6	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
7	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
8	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
9	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
10	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

 

Table No. I 

Summary of Clinical Signs of Toxicity and Mortality

Test System : Sprague Dawley Rat

Sex : Male

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No clinical signs observed	5	1 - 5	Day 0 - Day 14	0/5

 

Sex : Female 

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No clinical signs observed	5	6 - 10	Day 0 - Day 14	0/5

 

 Table No. II 

Summary of Evaluation of Dermal Reaction

Test System : Sprague Dawley Rat

Sex : Male 

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No dermal reaction observed	5	1 - 5	Day 0 - Day 14	0/5

 

 

 

Sex : Female

 

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No dermal reaction observed	5	6 - 10	Day 0 - Day 14	0/5

 

 

 

Table No.III 

Mean Body Weight and Percent Body Weight Gain (g)

Test System : Sprague Dawley Rat

Sex : Male

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	2000	Mean	244.22	292.20	19.64	315.82	8.08	29.31
		± SD	2.15	4.49	1.25	6.53	1.60	1.89

 

 

Sex : Female

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	2000	Mean	221.38	245.94	11.09	256.72	4.40	15.97
		± SD	2.67	4.63	1.31	3.02	1.12	0.97

 

  

Table No.IV 

Summary of Gross Pathological Findings

Test System : Sprague Dawley Rat

Sex : Male

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	2000	1 - 5	TS	No abnormality detected

 

Sex : Female

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	2000	6 - 10	TS	No abnormality detected

TS = Terminal Sacrifice

Interpretation of results:

other: Not irritating

Conclusions:

The test chemical was considered to be not irritating to the skin of rabbits.

Executive summary:

Various studies were performed on rabbits to assess the dermal irritation potential of test chemical which have been summarized as follows:

The skin irritation study of test chemical was conducted in Russian breed rabbits. Six rabbits (3 males/3 females) of the Russian breed, weighing 1.5 to 2 kgs, were shaved on the whole back and flanks two days before treatment with an electric clipper. The shaven skin on the left side was slightly scarified immediately before treatment. A gauze patch 2.5 x 2.5 cm was soaked with the test substance and immediately applied to the prepared skin. The patch was covered with a plastic film 5 x 5 cm, which was fixed, to the body with adhesive tape. The test chemical was applied to each side in quantities of 0.5 g (500mg). Before application a 50 % polyethylene glycol trituration (PEG 400) was made. The gauze patches were removed 24 hours after the application. The reaction of the skin was appraised upon removal and 72 hours after it. The primary irritation index as the measure of the acute skin irritation provoked by the substance was defined as the average of all the ratings found after 24 and 72 hours on the intact and slightly scarified skin. Compounds producing combined averages (primary irritation indexes) of 2 or less are only mildly irritating; whereas those with indexes from 2 to 5 are moderate irritants, and those with scores above 6 are considered severe irritants.

The primary irritation index of test chemical was evaluated to be 0.0 out of maximum score of 8. Since the treated rabbits did not develop any signs of skin irritation, the test chemical was considered to be not irritating to the skin.

In another skin irritation study which was designed and conducted to determine the dermal reaction profile of test chemical in Sprague Dawley rats. The study was performed as per OECD Guidelines 402 and complying to the GLP procedures.Ten rats (5 male and 5 female) were used for conducting dermal irritation/ corrosion study.The animals were kept in their cages for at least 5 days prior to administration for acclimatization to the laboratory condition and after acclimatization period, animals were randomly selected. Approximately 24 hours before application, the hair of each rat was closely clipped from the trunk (dorsal surface and sides from scapular to pelvic area) with an electric clipper, so as to expose at least 10% of the body surface area. The test item was applied directly onto the exposed skin of the animal, taking care to spread the test item evenly over the entire area of approximately 10% of the total body surface area or as much of the area as can reasonably be covered. The test item was held in contact with the skin using a porous gauze dressing and non irritating tape around the animal to cover the exposure site for first 24 hours exposure period. Elizabethan collar was placed on each animal for first 24 hours after application of the test item. These collars prevent ingestion of test item. Following 24 hours of exposure, the wrapping was removed and the test site wiped free of excess test item. Distilled water was used to remove residual test item. The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Also, the erythema and edema score of rats was calculated as 0. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment. Hence, it was concluded that test chemical was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested and Classified as “Category- Not Classified” as per CLP Classification.

All these studies lead 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”.

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:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data of test substances

Justification for type of information:

Data for the target chemical is summarized based on the available studies

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

WoE report is based on eye irritation studies as- WoE-2 and WoE-3
An eye irritation studies were condcuted on rabbits to assess its ocular effects.

GLP compliance:

not specified

Species:

other: 2.rabbit 3.rabbit

Strain:

other: 2.Russian breed 3.albino

Details on test animals or tissues and environmental conditions:

2.TEST ANIMALS
- Housing: The substance was tested on 6 rabbits (3 males/3 females) of.the Russian breed, which were kept separately in V2A wire cages
- Diet (e.g. ad libitum): fed on NAFAG, Gossau SG, rabbit food and water ad libitum
- Only rabbits showing normal ophthalmic findings were included in the test.

3.no data available

Vehicle:

other: 2.unchanged (no vehicle) 3.unchanged (no vehicle)

Controls:

yes

other: 2.The right eye served as a control

Amount / concentration applied:

2.
0.1 g

3.
3 mg

Duration of treatment / exposure:

2.
Amounts of 0.1 g of the test substance were introduced into the conjunctival sac of the left eye with a spatula. After application, the eyelids were held open for a few seconds. The treated eyes of three rabbits were each rinsed with 10 ml lukewarm water about 30 seconds after treatment.

3.
single exposure

Observation period (in vivo):

2.The reactions were appraised with a slit-lamp after 24 hours and after 2, 3, 4 and 7 days
3.
24,48 and 72 hours

Duration of post- treatment incubation (in vitro):

no data available

Number of animals or in vitro replicates:

2.
6 (3 males/3 females)

3.
6

Details on study design:

2.REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated eyes of three rabbits were each rinsed with 10 ml lukewarm water about 30 seconds after treatment.

Procedure : Amounts of 0.1 g of the test substance were introduced into the conjunctival sac of the left eye with a spatula.After application, the eyelids were held open for a few seconds. The right eye served as a control. The treated eyes of three rabbits were each rinsed with 10 ml lukewarm water about 30 seconds after treatment.
TOOL USED TO ASSESS SCORE: slit lamp.

3.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no

SCORING SYSTEM: Irritative effects observed in the eye were scored according to the Draize method.

Irritation parameter:

overall irritation score

Remarks:

2.

Basis:

mean

Time point:

other: 7 days

Score:

0

Max. score:

110

Reversibility:

not specified

Remarks on result:

other: score is 0 for cornea, iris, conjuctiva

Irritation parameter:

overall irritation score

Remarks:

3.

Basis:

mean

Time point:

72 h

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

2.
The irritation index was found to be 0 for the cornea, 0 for the iris and 0 for the conjunctivae.
The total maximum is 80 for the cornea, 10 for the iris and 20 for the conjunctivae.

3.
No gross signs of eye irritation were observed at any observation interval following application of the test chemical to albino rabbits. All rabbits showed normal appearance throughout the study and there was no evidence of systemic toxicity from mucous membrane absorption.

Interpretation of results:

other: not irritating

Conclusions:

The Test chemical was considered to be not irritating to the eyes of treated rabbits.

Executive summary:

Various studies were performed on rabbits to assess the ocular irritation potential of test chemical which have been summarized as follows:

An eye irritation study of test chemical was conducted in six (3 males/3 females) Russian breed rabbits. Amounts of 0.1 g of the test substance were introduced into the conjunctival sac of the left eye with a spatula. After application, the eyelids were held open for a few seconds. The treated eyes of three rabbits were each rinsed with 10 ml lukewarm water about 30 seconds after treatment. The reactions were appraised with a slit-lamp after 24 hours and after 2, 3, 4 and 7 days. The total maximum is 80 for the cornea, 10 for the iris and 20 for the conjunctivae The irritation index was estimated to be 0 for the cornea, 0 for the iris and 0 for the conjunctivae out of maximum score of 110. Since there were no evidence of any signs of ocular lesions, the test chemical was considered to be not irritating to the rabbits’ eye.

In another eye irritation study which was carried out on rabbits to assess the irritation potency of test chemical. 3 mg of undiluted test chemical was instilled into the eyes of 6 albino rabbits. The untreated eye served as control. The eyes remained unwashed throughout the test.Each rabbit was examined for signs of irritation and for gross signs of systemic toxicity at intervals of 24, 48 and 72hours after application. The effects observed were scored according to Draize method.No gross signs of eye irritation were observed at any observation interval following application of the test chemical to albino rabbits. All rabbits showed normal appearance throughout the study and there was no evidence of systemic toxicity from mucous membrane absorption.Hence,test chemical can be considered not irritating to rabbit eyes.

The results obtained from these studies lead to a conclusion that Test chemical is indeed not irritating to eye. Hence, comparing the above annotations with the criteria of CLP regulation, Test chemical can be classified under the category “Not Classified”.

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 were performed on rabbits to assess the dermal irritation potential of test chemical which have been summarized as follows:

The skin irritation study of test chemical was conducted in Russian breed rabbits. Six rabbits (3 males/3 females) of the Russian breed, weighing 1.5 to 2 kgs, were shaved on the whole back and flanks two days before treatment with an electric clipper. The shaven skin on the left side was slightly scarified immediately before treatment. A gauze patch 2.5 x 2.5 cm was soaked with the test substance and immediately applied to the prepared skin. The patch was covered with a plastic film 5 x 5 cm, which was fixed, to the body with adhesive tape. The test chemical was applied to each side in quantities of 0.5 g (500mg). Before application a 50 % polyethylene glycol trituration (PEG 400) was made. The gauze patches were removed 24 hours after the application. The reaction of the skin was appraised upon removal and 72 hours after it. The primary irritation index as the measure of the acute skin irritation provoked by the substance was defined as the average of all the ratings found after 24 and 72 hours on the intact and slightly scarified skin. Compounds producing combined averages (primary irritation indexes) of 2 or less are only mildly irritating; whereas those with indexes from 2 to 5 are moderate irritants, and those with scores above 6 are considered severe irritants.

The primary irritation index of test chemical was evaluated to be 0.0 out of maximum score of 8. Since the treated rabbits did not develop any signs of skin irritation, the test chemical was considered to be not irritating to the skin.

In another skin irritation study which was designed and conducted to determine the dermal reaction profile of test chemical in Sprague Dawley rats. The study was performed as per OECD Guidelines 402 and complying to the GLP procedures.Ten rats (5 male and 5 female) were used for conducting dermal irritation/ corrosion study.The animals were kept in their cages for at least 5 days prior to administration for acclimatization to the laboratory condition and after acclimatization period, animals were randomly selected. Approximately 24 hours before application, the hair of each rat was closely clipped from the trunk (dorsal surface and sides from scapular to pelvic area) with an electric clipper, so as to expose at least 10% of the body surface area. The test item was applied directly onto the exposed skin of the animal, taking care to spread the test item evenly over the entire area of approximately 10% of the total body surface area or as much of the area as can reasonably be covered. The test item was held in contact with the skin using a porous gauze dressing and non irritating tape around the animal to cover the exposure site for first 24 hours exposure period. Elizabethan collar was placed on each animal for first 24 hours after application of the test item. These collars prevent ingestion of test item. Following 24 hours of exposure, the wrapping was removed and the test site wiped free of excess test item. Distilled water was used to remove residual test item. The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Also, the erythema and edema score of rats was calculated as 0. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment. Hence, it was concluded that test chemical was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested and Classified as “Category- Not Classified” as per CLP Classification.

All these studies lead 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”.

Eye irritation:

Various studies were performed on rabbits to assess the ocular irritation potential of test chemical which have been summarized as follows:

An eye irritation study of test chemical was conducted in six (3 males/3 females) Russian breed rabbits. Amounts of 0.1 g of the test substance were introduced into the conjunctival sac of the left eye with a spatula. After application, the eyelids were held open for a few seconds. The treated eyes of three rabbits were each rinsed with 10 ml lukewarm water about 30 seconds after treatment. The reactions were appraised with a slit-lamp after 24 hours and after 2, 3, 4 and 7 days. The total maximum is 80 for the cornea, 10 for the iris and 20 for the conjunctivae The irritation index was estimated to be 0 for the cornea, 0 for the iris and 0 for the conjunctivae out of maximum score of 110. Since there were no evidence of any signs of ocular lesions, the test chemical was considered to be not irritating to the rabbits’ eye.

In another eye irritation study which was carried out on rabbits to assess the irritation potency of test chemical. 3 mg of undiluted test chemical was instilled into the eyes of 6 albino rabbits. The untreated eye served as control. The eyes remained unwashed throughout the test.Each rabbit was examined for signs of irritation and for gross signs of systemic toxicity at intervals of 24, 48 and 72hours after application. The effects observed were scored according to Draize method.No gross signs of eye irritation were observed at any observation interval following application of the test chemical to albino rabbits. All rabbits showed normal appearance throughout the study and there was no evidence of systemic toxicity from mucous membrane absorption.Hence,test chemical can be considered not irritating to rabbit eyes.

The results obtained from these studies lead to a conclusion that Test chemical is indeed not irritating to eye. Hence, comparing the above annotations with the criteria of CLP regulation, Test chemical can be classified under the category “Not Classified”.

Justification for classification or non-classification

The skin and eye irritation potential of test chemical was observed in various studies. The results obtained from these studies indicate that the chemical is not likely to cause neither skin irritation nor irritant to eye. Hence the test chemical can be classified under the category “Not Classified” for skin irritation and eye as per CLP.