Hydrogen tetrasodium bis[2-[[6-[[4-chloro-6-[3-sulphoanilino]-1,3,5-triazin-2-yl]amino]-1-hydroxy-3-sulpho-2-naphthyl]azo]benzoato(4-)]chromate(5-)

Skin irritation / corrosion

Administrative data

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

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from experimental study report

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Principles of method if other than guideline:

Acute dermal irritation study was performed to determine the irritant nature of the test chemical

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

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

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

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.

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

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.

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24 hrs

Score:

0.67

Max. score:

0

Reversibility:

fully reversible

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24 hrs

Score:

0

Max. score:

0

Reversibility:

fully reversible

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 48 hrs

Score:

0.33

Max. score:

0

Reversibility:

fully reversible

Irritation parameter:

edema score

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

Irritant / corrosive response data:

No erythema and edema (skin irritation) were found at the end of 72 hour observation period after patch removal.

Other effects:

no effects observed

Table 1

Skin Reaction

 

In Treated area Dose : 0.5 g (Pulverized form)                                                     Sex:Male

 

Animal No.	Test	Treated  area	Erythema score				Oedema score
			1h	24h	48h	72h	1h	24h	48h	72h
1	Initial	Right	1	1	1	0	0	0	0	0
2	Confirmatory	Left	1	1	0	0	0	0	0	0
3		Right	1	1	1	0	0	0	0	0

 

 

 

 

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

 

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)

Table 1 Continued…

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

 

Animal Number                   Observations	1	2	3
Erythema	0.67	0.33	0.67
Oedema	0.00	0.00	0.00

Table 2

Individual Animal BodyWeight

Sex:Male

Animal No.	Body Weight (kilogram)
	Prior to Dosing	At termination
1	2.090	2.178
2	2.144	2.172
3	2.354	2.386

Table 3

Individual AnimalClinicalSigns

Sex:Male

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:

not irritating

Conclusions:

No erythema and no oedema (skin irritation) were found at the end of 72 hour observation period after patch removal. Hence, it was concluded that the test chemical was "Non-Irritating" to the skin of 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.

Executive summary:

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.

Cross-reference 2

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from experimental report

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Principles of method if other than guideline:

The test compound was tested for its dermal irritation potential in New Zealand White Rabbits according to OECD guideline 404 Guidelines

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

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 bottles, ad libitum
Acclimatization: The healthy rabbits selected for study was acclimatized to standard laboratory condition for one week in experimental room under Veterinary examination.
Randomization: After acclimatization and Veterinary examination three females were randomly selected.

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 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,mad-libitum

Type of coverage:

occlusive

Preparation of test site:

clipped

Remarks:

intact skin

Vehicle:

water

Controls:

not specified

Amount / concentration applied:

0.5 gm was moistened with distilled water

Duration of treatment / exposure:

4 hours

Observation period:

60 min., 24, 48 and 72 hours after application.

Number of animals:

3 female rabbits

Details on study design:

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

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

14 d

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

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.

Other effects:

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

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

Interpretation of results:

other: not irritating

Conclusions:

The Primary Irritation Index (PII) for Test chemical after 14 days of observation was 0.0. Also 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.

Executive summary:

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.

Cross-reference 3

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from experimental study report

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

To determine the dermal reaction profile of test chemical in Sprague Dawley rat

GLP compliance:

yes

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

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:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

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:

24 hours

Observation period:

14 days

Number of animals:

10 (5/sex).

Details on study design:

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:

erythema score

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

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:

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:

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 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.

Individual Animal - Evaluation of Dermal Reaction

 

Laboratory Test Item Code :TAS/122/013

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	269.94	296.10	9.69	314.52	6.22	16.50
		± SD	2.32	6.07	1.61	7.10	1.18	1.65

 

 

 

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	244.56	256.84	5.02	266.86	3.89	9.12
		± SD	4.41	5.32	1.27	6.98	0.78	2.08

 

 

 

  

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 overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) were observed at the end of 14 days after patch removal. Hence, it was concluded that the test substance was not-Irritating to the skin of rats under the experimental conditions tested .Thus it can be concluded that the substance can be considered as not irritating.

Executive summary:

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.

Data source

Reference

Reference Type:

publication

Title:

WoE of dermal irritation study for CAS no 73507-17-2

Author:

Sustainability Support Services (Europe) AB

Year:

2019

Bibliographic source:

WoE report, Sustainability Support Services (Europe) AB, 2019

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

Hydrogen tetrasodium bis[2-[[6-[[4-chloro-6-[3-sulphoanilino]-1,3,5-triazin-2-yl]amino]-1-hydroxy-3-sulpho-2-naphthyl]azo]benzoato(4-)]chromate(5-)

EC Number:

277-492-0

EC Name:

Hydrogen tetrasodium bis[2-[[6-[[4-chloro-6-[3-sulphoanilino]-1,3,5-triazin-2-yl]amino]-1-hydroxy-3-sulpho-2-naphthyl]azo]benzoato(4-)]chromate(5-)

Cas Number:

73507-17-2

Molecular formula:

C52H28Cl2CrN14O18S44Na

IUPAC Name:

Hydrogen tetrasodium bis[2-[[6-[[4-chloro-6-[3-sulphoanilino]-1,3,5-triazin-2-yl]amino]-1-hydroxy-3-sulpho-2-naphthyl]azo]benzoato(4-)]chromate(5-)

Details on test material:

- Name of the test chemical: Hydrogen tetrasodium bis[2-[[6-[[4-chloro-6-[3-sulphoanilino]-1,3,5-triazin-2-yl]amino]-1-hydroxy-3-sulpho-2-naphthyl]azo]benzoato(4-)]chromate(5-)
- Molecular formula: C52H28Cl2CrN14O18S4.H.4Na
- Molecular weight: 2693.61 g/mol
- Substance type: Organic

Test animals

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

Test system

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.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Remarks:

2

Basis:

mean

Time point:

other: 24 hrs

Score:

0.67

Max. score:

0

Reversibility:

fully reversible

Results 2

Irritation parameter:

edema score

Remarks:

2

Basis:

mean

Time point:

other: 24 hrs

Score:

0

Max. score:

0

Reversibility:

fully reversible

Results 3

Irritation parameter:

erythema score

Remarks:

2

Basis:

mean

Time point:

other: 48 hrs

Score:

0.33

Max. score:

0

Reversibility:

fully reversible

Results 4

Irritation parameter:

edema score

Remarks:

2

Basis:

mean

Time point:

other: 48 hrs

Score:

0

Max. score:

0

Reversibility:

fully reversible

Results 5

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 72 hrs

Score:

0

Max. score:

0.67

Reversibility:

fully reversible

Results 6

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 72 hrs

Score:

0

Max. score:

0

Reversibility:

fully reversible

Results 7

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

Results 8

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

Results 9

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.

Applicant's summary and conclusion

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.