Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Skin irritation:

No erythema and no oedema (skin irritation) were found at the end of observation period after patch removal. Hence, it was concluded that the test chemical was "Non-Irritating" to the skin of New Zealand White rabbits under the experimental conditions tested and hence the test chemical is "Not Classified as Skin Irritant" as per CLP regulation.

Eye irritation:

The test chemical did not induce ocular irritation in the eyes of rabbits. Hence the test chemical is "Not Classified as Eye Irritant" as per CLP regulation.

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:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Experimental data from various test chemicals
Justification for type of information:
Data for the target chemical is summarized based on data from various test chemicals
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Principles of method if other than guideline:
WoE for the target chemical is summarized based on experimental data from various test chemicals
GLP compliance:
not specified
Species:
other: 2/3. rabbit 4. rat
Strain:
other: 2./3. New Zealand White 4. Sprague-Dawley
Details on test animals or test system and environmental conditions:
2. Species : Rabbit (Oryctolagus cuniculus)
Strain : New Zealand White
Age : 3.5 to 4.7 Months (Approximately)
Sex : Male
Number of Animals : Three
Supplier/Source : Procured from GENTOX BIOSERVICES PVT. LTD.
Health Status : Healthy young adult rabbits were used for the study
Body weight of animals : Minimum: 2.090 kg & Maximum: 2.354 kg (Prior to Treatment)
Acclimatisation :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.
Identification : During acclimatization marking was done with non toxic marker pen in the inside of left ear of rabbits and after acclimatization, animals were marked with permanent number in the inner side of right ear of rabbits. Permanent marker and cage card was used for identification. The individual cage cards were labelled with study no., study type, test system, sex, dose, animal number, experimental start and completion date.
Husbandry Conditions
Diet : All animals were provided conventional laboratory rabbit diet
Water : Aqua guard filtered tap water was provided ad libitum.
Husbandry : 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.
Experimental Room Condition
Temperature : Minimum: 18.70 °C Maximum: 21.60 °C
Relative humidity : Minimum: 49.10% Maximum: 69.40%
Light-dark-rhythm : 12 hour light and 12 hour dark
Air Changes : More than 12 changes per hour

3. Details on test animals
Age: 10 to 12 weeks
Sex:Female
Body weight range: 2.0kg±200g
Identification : By cage tag and corresponding colour body marking
Housing:Animals were housed individually in stainless steel cages provided with stainless steel mesh bottom and facilities for food and water bottle.
Diet:Pelleted feed supplied by Pranav agro Industries Ltd., B7/6 Ramesh Nagar, Delhi Water:Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bott les, ad libitum
Acclimatization: The healthy rabbits selected for study was acclimatized to standard laboratory condi tion for one week in experimental room under Veterinary examination.
Randomization: After acclimatization and Veterinary examination three females were randomly s elected.
Details on environmental conditions:
- Temperature (°C): temperature between 22-25 deg C
- 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.

HUSBANDRY
Environmental conditions :Air conditioned rooms with 10-15 air changes per hour, temperature be tween 19-25 0C, relative humidity 30-60% and illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark.

Accommodation Animals were housed individually in stainless steel cages provided with stainless s teel mesh bottom and facilities for food and water bottle.
Diet : Pelleted feed supplied by Pranav agro Industries Ltd., B7/6 Ramesh Nagar, Delhi, India Water : Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles,mad-libitum

4. 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 6 – 9 weeks were used.
- Weight at study initiation: The weight ranges of approximately 239.1 to 272.5 grams at initiation of dosing were used.
Body weights at the start :
Male
Mean : 269.94 g (= 100 %)
Minimum : 266.4 g (- 1.31 %)
Maximum : 272.5 g (+ 0.95 %)
Total No. of animals : 5

Female
Mean : 244.56 g (= 100 %)
Minimum : 239.1 g (- 2.23 %)
Maximum : 250.4 g (+ 2.39 %)
Total No. of animals : 5
- Fasting period before study: No data
- 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): 20.1 to 22.3 degree centigrade.
- Humidity (%): 55.7% to 59.6%.
- 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: 30-09-2016 to 15-10-2016
Type of coverage:
other: 2. semiocclusive 3. occlusive 4. semiocclusive
Preparation of test site:
clipped
Remarks:
clipped intact skin
Vehicle:
other: 2. unchanged (no vehicle) 3. water 4. unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
2. Preparation of Application Site
Approximately 24 h prior to treatment, the fur coat of each rabbit was removed from dorsal lumbar region approximately 6 X 6 cm at contralateral sites on each rabbit using clipper, one as control and other site as treatment. Rabbits with healthy intact skin were selected for the study.

Test Item Application Procedure
The 0.5 ml of test item (as such) was applied uniformly over clipped area (approximately 6 X 6 cm) of the trunk of each rabbit skin (treated site) and 0.5 ml distilled water was applied at control site. Test item was held in contact with the skin with a porous gauze dressing and non-irritating tape (Micropore 3”) throughout a 4-hour exposure period, to prevent access by the rabbits to the patch and resultant ingestion of the test item. At the end of the exposure period, residual test item was removed by using cotton soaked in distilled water.
A single rabbit (Animal No. 1) was used for initial testing. The patch was removed after 4 h and the responses graded, no erythema and oedema was observed at 1 hour, 24 hour, 48 hour and 72 hour in animal no. 1. Hence the confirmatory test was conducted on additional two rabbits (No. 2 and 3) after 24 hour to confirm the non irritant nature of the test item.

3. 0.5 gm was moistened with distilled water

4. 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
Duration of treatment / exposure:
2./3. 4 hours
4. 24 hrs
Observation period:
2. 72 hours
3. 60 min., 24, 48 and 72 hours after application
4. 14 days
Number of animals:
2. 3
3. 3 female rabbits
4. 10 (5/sex)
Details on study design:
2. A dose of 0.5 gm of test item moistened with 0.5 ml of distilled water was applied to the skin, over an area of approximately 6 x 6 cm clipped of hair on one side of rabbits.

3. TEST SITE - Area of exposure: dorsal area of trunk - % coverage: small area (approximately 6 cm2) - Type of wrap if used: impervious dressing which was secured in position with adhesive tape

REMOVAL OF TEST SUBSTANCE - Washing (if done): yes - Time after start of exposure:after patch removal the test site was washed with lukewarm water

OBSERVATION TIME POINTS (indicate if minutes, hours or days) : The intact skin site of application of each animal was observed for signs of erythema and oedema at 60 min., 24, 48 and 72 hours after application.

SCORING SYSTEM: - Method of calculation:The intact skin site of application of each animal was observed for signs of erythema and oedema and the responses were scored following Draize’s method

4. 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 observ ations were conducted at 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily t hereafter 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:
erythema score
Remarks:
2
Basis:
mean
Time point:
other: 24 hrs
Score:
0.67
Max. score:
0
Reversibility:
fully reversible
Irritation parameter:
edema score
Remarks:
2
Basis:
mean
Time point:
other: 24 hrs
Score:
0
Max. score:
0
Reversibility:
fully reversible
Irritation parameter:
erythema score
Remarks:
2
Basis:
mean
Time point:
other: 48 hrs
Score:
0.33
Max. score:
0
Reversibility:
fully reversible
Irritation parameter:
edema score
Remarks:
2
Basis:
mean
Time point:
other: 48 hrs
Score:
0
Max. score:
0
Reversibility:
fully reversible
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 72 hrs
Score:
0
Max. score:
0.67
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 72 hrs
Score:
0
Max. score:
0
Reversibility:
fully reversible
Irritation parameter:
primary dermal irritation index (PDII)
Remarks:
3
Basis:
mean
Time point:
14 d
Score:
0
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Remarks:
4
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:
4
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:
2. No erythema and edema (skin irritation) were found at the end of 72 hour observation period after patch removal.

3. The test compound applied at the dose level of 500mg on shaven back skin of rabbit did not produced any irritation to skin during period of observation.

4. 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:
2. no effects observed

3. The test compound applied on the shaven back skin of rabbit did not produce pain and any clinical si gns of toxicity throughout the examination period of 14 days.

4. 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 dur ing 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 d uring 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 9.69% and 16.50% respectively.

Sex : Female
Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 5.02% and 9.12% respectively.

Gross Pathological Findings Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group.
Interpretation of results:
not irritating
Conclusions:
No erythema and no oedema (skin irritation) were found at the end of observation period after patch removal. Hence, it was concluded that the test chemical was "Non-Irritating" to the skin of animals under the experimental conditions tested and hence the test chemical is "Not Classified as Skin Irritant" as per CLP regulation.
Executive summary:

Data available for the various test chemicals was reviewed to determine the toxic nature of the test chemical. The studies are as mentioned below:

A dermal irritation study was conducted on New Zealand white rabbits in accordance with OECD 404 to assess the irritation parameter of Test chemical. Three male New Zealand White rabbits were used for the study. The hair of al the rabbits were clipped at contraleteral sites, approximately 24 hrs prior to treatment. A dose of 0.5g of pulverised form of test chemical moistened with 0.5mL distilled water was applied to the skin over an area of 6X6 cm clipped of hair on one side of rabbits. The other untreated site served as control area and 0.5mL distilled water was applied at this site. At the end of 4hrs, the gauze patch was removed and test item application site was wiped with water without altering the epidermis. Initially the test chemical was applied to the clipped skin of one rabbit. The test site was covered with gauze patch. After 4 hrs of exposure, very slight erythema and no oedema was observed at 1 hr of observation. At 24 and 48 hrs, very slight erythema and no oedema was observed. At 72 hrs, no erythema and no oedema was observed. Hence the confirmatory test was conducted on two more animals. The patch was removed at 4hrs and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hrs after patch removal, evaluated and graded as per draize method. In the animal no. 2 and 3 at 1 and 24 hrs observation post patch removal, revelaed very slight erythema and no oedema. At 48 hrs, animal no. 2 was observed with no erythema and no oedema whereas animal no. 3 was observed with slight erythema and no oedema. At 72 hrs, no erythema and no oedema was observed in animals no. 2 and 3. The other untreated site revealed no erythema and no oedema and was found to be normal throughout the experimental period. The individual mean score at treated site at 24, 48 and 72 hrs for animals 1, 2, 3 were 0.67, 0.33, 0.67 and 0.00, 0.00, 0.00 for erythema and oedema formation, respectively. At 72 hrs, no erythema and no oedema was observed after patch removal. 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 hence the test chemical is "Not Classified as Skin Irritant" as per CLP regulation.

A dermal irritation study was conducted on New Zealand white rabbits in accordance with OECD 404 to assess the irritation parameter of Test chemical. Three female New Zealand White rabbits.    In the initial test one healthy rabbit of body weight 2.04 kg selected for study after acclimatization. The test compound in the amount of 0.5 gm was applied uniformly at the different sites on the shaven back skin of animal. The hairs of back sides were removed (approximately 6 cm2) one day earlier before the treatment. 0.5 gm was moistened with distilled water and then applied to a small area (approximately 6 cm2) of skin and covered with a gauze patch, which was held in place with non-irritating tape. The first patch was applied on the shaven back skin of rabbit and removed after three minutes. No serious reaction was observed at the site of application. The second patch was applied on the different shaven back side and removed after one hour. There were no signs of skin reaction observed at this site of application. Finally, a third patch was applied at a different site and was removed after four hour. No dermal irritation was observed at the site of application of the test compound after 4 hours of patch removal. Finally, the animal was observed for 14 days, for any irritation and corrosion.  Because of there was no corrosive effect was observed in the initial test, a confirmatory test was done in order to confirm the irritant or negative response of the test substance by using two additional animals. In the confirmatory test the test compound in the amount of 0.5 gm was applied on the shaven back skin (approximately 6 cm2) of two animals (body weight of each animal was 2.17 and 2.13kg), each with one patch, for an exposure period of four hours. After four hours the patch was removed and the skin reactions were graded according to Draize’s method.  The Primary Irritation Index (PII) for Test chemical after 14 days of observation was 0.0. Also the test chemical did not produce pain and 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 and hence it is classified in Category "Not classified" as per the criteria mention in CLP regulation.

A study 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 the test substance was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested.

No erythema and no oedema (skin irritation) were found at the end of observation period after patch removal. Hence, it was concluded that the test chemical was "Non-Irritating" to the skin of New Zealand White rabbits under the experimental conditions tested and hence the test chemical is "Not Classified as Skin Irritant" as per CLP regulation.

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:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Experimental data from various test chemicals
Justification for type of information:
Data for the target chemical is summarized based on data from various test chemicals
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Principles of method if other than guideline:
WoE for the target chemical is summarized based on experimental data from various test chemicals
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Remarks:
3. Not specified
Details on test animals or tissues and environmental conditions:
2. Details on test animals:
- Age: 10 to 12 weeks
- Sex: Female
- Body weight range: 2.0kg ±200g
- Housing: Rabbit was housed in stainless steel cages provided with stainless steel mesh bottom and facilities for food and water bottle.
- Diet:Pelleted feed supplied by Pranav agro Industries Ltd., B7/6 Ramesh Nagar, Delhi, India
- Water:Community tap water passed through ‘Aqua Guard on line water filter’ was kept in bottles, ad libitum.
- Acclimatization: The rabbit was acclimatized to standard laboratory condition for 24 hours in experimental room before study.

ENVIRONMENTAL CONDITIONS
- Temperature:temperature between 22-250C
- Humidity (%):relative humidity 40-60%
- Air changes (per hr):Air conditioned room 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.


HUSBANDRY
Environmental conditions :Air conditioned rooms with 10-15 air changes per hour, temperature between 19-25 0C, relative humidity 30-60% and illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark.
Accommodation Animals were housed individually in stainless steel cages provided with stainless steel mesh bottom and facilities for food and water bottle.
Diet : Pelleted feed supplied by Pranav agro Industries Ltd., B7/6 Ramesh Nagar, Delhi, India
Water : Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles,ad-libitum

3. No data
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
100mg (0.1g)
Duration of treatment / exposure:
2. 24 hours
3. No data
Observation period (in vivo):
2. The eyes were examined at 1, 24, 48 and 72 hours after test substance application
3. No data
Duration of post- treatment incubation (in vitro):
no data available
Number of animals or in vitro replicates:
2. 3 female rabbits
3. No data
Details on study design:
2. REMOVAL OF TEST SUBSTANCE
- Washing (if done): not washed
SCORING SYSTEM:Scale of weighted scores for grading the severity of ocular lesions developed by Draize et al
TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein: hand-slit lamp
Examination of reactions was facilitated by use of biomicroscope and hand slit lamp. After recording the observations at 24 hours, the eyes were further examined with the aid of fluorescein.

3. No data
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
72 h
Score:
0
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
2
Irritation parameter:
overall irritation score
Basis:
mean
Remarks on result:
no indication of irritation
Remarks:
3
Irritant / corrosive response data:
2. The test compound when applied to the eye of New Zealand white rabbit at the dose level of 0.1gm did not produce any lesions such as pannus, staining throughout the observation period of 72 hours.
Other effects:
2. The test compound applied in conjunctival sac of rabbits did not show any observable clinical signs of eye irritation throughout the observation period of 21 days.
Interpretation of results:
other: not irritating
Conclusions:
The test chemical did not induce ocular irritation in the eyes of rabbits. Hence the test che mical is likely to be "Non irritating" to rabbit eyes.
Executive summary:

Data available for the various test chemicals was reviewed to determine the toxic nature of the test chemical. The studies are as mentioned below:

An ocular irritation study was conducted on New Zealand white rabbits in accordance with OECD 405 to assess the irritation parameter of the test chemical. Three female New Zealand White rabbits were used for the study. In the initial test, One healthy rabbit of body weight 2.11 kg was selected for study after acclimatization. Both eyes of rabbit were examined for any abnormal discharge such as eye irritation, ocular defects or pre-existing corneal injury from eye 24 hours prior to application of test compound. 0.1 gm of the test chemical was applied by gently pulling the eye lids into the conjunctival sac of the rabbit.The lids were then gently held for about one second in order to prevent loss of the material. The other eye which remain untreated, served as a control. The acute irritation to eye conjunctiva, cornea and iris was evaluated at 1, 24, 48 and 72 hoursafter the treatment. The grades of ocular reaction (conjunctiva, cornea and iris) were recorded at each observation. To determine the reversibility of the effect the animal was 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 weight before and during the study was observed. The test compound when applied to conjunctival sac of rabbit in the amount of 0.1 gm did not produce any eye irritation or any eye discharge. Furthermore, no other clinical signs were recorded after application of test compound such as cage side activity, pain etc. The result obtained from the initial test was confirmed in additional two animal (body weight ranges 200±gm) of same sex and same dose level. The acute irritation to eye conjunctiva, cornea and iris was evaluated at 1, 24, 48 and 72 hoursafter the treatment. The grades of ocular reaction (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. The overall irritation index of Test chemical was 0.0 after 72 hours. Also 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.

Ocular irritation study was performed to determine the toxic nature of the test chemical. The study was performed by administering 100 mg of the test chemical to 3 male rabbits. Based on the reported results, the test chemical did not induce ocular irritation in the eyes of 3 male rabbits. Hence the test chemical is likely to be "Non irritating".

Based on the data available and applying the weight of evidence approach, the test chemical did not induce ocular irritation in the eyes of rabbits. Hence the test che mical is likely to be "Non irritating" to rabbit eyes.

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

Additional information

Skin irritation:

Data available for the various test chemicals was reviewed to determine the toxic nature of the test chemical. The studies are as mentioned below:

A dermal irritation study was conducted on New Zealand white rabbits in accordance with OECD 404 to assess the irritation parameter of Test chemical. Three male New Zealand White rabbits were used for the study. The hair of al the rabbits were clipped at contraleteral sites, approximately 24 hrs prior to treatment. A dose of 0.5g of pulverised form of test chemical moistened with 0.5mL distilled water was applied to the skin over an area of 6X6 cm clipped of hair on one side of rabbits. The other untreated site served as control area and 0.5mL distilled water was applied at this site. At the end of 4hrs, the gauze patch was removed and test item application site was wiped with water without altering the epidermis. Initially the test chemical was applied to the clipped skin of one rabbit. The test site was covered with gauze patch. After 4 hrs of exposure, very slight erythema and no oedema was observed at 1 hr of observation. At 24 and 48 hrs, very slight erythema and no oedema was observed. At 72 hrs, no erythema and no oedema was observed. Hence the confirmatory test was conducted on two more animals. The patch was removed at 4hrs and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hrs after patch removal, evaluated and graded as per draize method. In the animal no. 2 and 3 at 1 and 24 hrs observation post patch removal, revelaed very slight erythema and no oedema. At 48 hrs, animal no. 2 was observed with no erythema and no oedema whereas animal no. 3 was observed with slight erythema and no oedema. At 72 hrs, no erythema and no oedema was observed in animals no. 2 and 3. The other untreated site revealed no erythema and no oedema and was found to be normal throughout the experimental period. The individual mean score at treated site at 24, 48 and 72 hrs for animals 1, 2, 3 were 0.67, 0.33, 0.67 and 0.00, 0.00, 0.00 for erythema and oedema formation, respectively. At 72 hrs, no erythema and no oedema was observed after patch removal. 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 hence the test chemical is "Not Classified as Skin Irritant" as per CLP regulation.

A dermal irritation study was conducted on New Zealand white rabbits in accordance with OECD 404 to assess the irritation parameter of Test chemical. Three female New Zealand White rabbits.    In the initial test one healthy rabbit of body weight 2.04 kg selected for study after acclimatization. The test compound in the amount of 0.5 gm was applied uniformly at the different sites on the shaven back skin of animal. The hairs of back sides were removed (approximately 6 cm2) one day earlier before the treatment. 0.5 gm was moistened with distilled water and then applied to a small area (approximately 6 cm2) of skin and covered with a gauze patch, which was held in place with non-irritating tape. The first patch was applied on the shaven back skin of rabbit and removed after three minutes. No serious reaction was observed at the site of application. The second patch was applied on the different shaven back side and removed after one hour. There were no signs of skin reaction observed at this site of application. Finally, a third patch was applied at a different site and was removed after four hour. No dermal irritation was observed at the site of application of the test compound after 4 hours of patch removal. Finally, the animal was observed for 14 days, for any irritation and corrosion.  Because of there was no corrosive effect was observed in the initial test, a confirmatory test was done in order to confirm the irritant or negative response of the test substance by using two additional animals. In the confirmatory test the test compound in the amount of 0.5 gm was applied on the shaven back skin (approximately 6 cm2) of two animals (body weight of each animal was 2.17 and 2.13kg), each with one patch, for an exposure period of four hours. After four hours the patch was removed and the skin reactions were graded according to Draize’s method.  The Primary Irritation Index (PII) for Test chemical after 14 days of observation was 0.0. Also the test chemical did not produce pain and 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 and hence it is classified in Category "Not classified" as per the criteria mention in CLP regulation.

A study 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 the test substance was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested.

No erythema and no oedema (skin irritation) were found at the end of observation period after patch removal. Hence, it was concluded that the test chemical was "Non-Irritating" to the skin of New Zealand White rabbits under the experimental conditions tested and hence the test chemical is "Not Classified as Skin Irritant" as per CLP regulation.

Eye irritation:

Data available for the various test chemicals was reviewed to determine the toxic nature of the test chemical. The studies are as mentioned below:

An ocular irritation study was conducted on New Zealand white rabbits in accordance with OECD 405 to assess the irritation parameter of the test chemical. Three female New Zealand White rabbits were used for the study. In the initial test, One healthy rabbit of body weight 2.11 kg was selected for study after acclimatization. Both eyes of rabbit were examined for any abnormal discharge such as eye irritation, ocular defects or pre-existing corneal injury from eye 24 hours prior to application of test compound. 0.1 gm of the test chemical was applied by gently pulling the eye lids into the conjunctival sac of the rabbit.The lids were then gently held for about one second in order to prevent loss of the material. The other eye which remain untreated, served as a control. The acute irritation to eye conjunctiva, cornea and iris was evaluated at 1, 24, 48 and 72 hoursafter the treatment. The grades of ocular reaction (conjunctiva, cornea and iris) were recorded at each observation. To determine the reversibility of the effect the animal was 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 weight before and during the study was observed. The test compound when applied to conjunctival sac of rabbit in the amount of 0.1 gm did not produce any eye irritation or any eye discharge. Furthermore, no other clinical signs were recorded after application of test compound such as cage side activity, pain etc. The result obtained from the initial test was confirmed in additional two animal (body weight ranges 200±gm) of same sex and same dose level. The acute irritation to eye conjunctiva, cornea and iris was evaluated at 1, 24, 48 and 72 hoursafter the treatment. The grades of ocular reaction (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. The overall irritation index of Test chemical was 0.0 after 72 hours. Also 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.

Ocular irritation study was performed to determine the toxic nature of the test chemical. The study was performed by administering 100 mg of the test chemical to 3 male rabbits. Based on the reported results, the test chemical did not induce ocular irritation in the eyes of 3 male rabbits. Hence the test chemical is likely to be "Non irritating".

Based on the data available and applying the weight of evidence approach, the test chemical did not induce ocular irritation in the eyes of rabbits. Hence the test chemical is "Not Classified as Eye Irritant" as per CLP regulation.

Justification for classification or non-classification

Based on the data available and applying the weight of evidence approach, the test chemical is not irritating to the skin and eyes. Hence the test chemical is "Not Classified as Skin and Eye Irritant" as per CLP regulation.