4'-aminoazobenzene-4-sulphonic acid

Endpoint summary

Description of key information

Skin irritation

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the dermal irritation potential of the test chemical. Based on the summarized studies for target chemical and its structurally and functionally similar read across substances,it can be concluded that the testchemical is unable to cause skin irritation and considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

Eye irritation

The ocular irritation potential of target chemical was assessedin various experimental studies which were conducted on rabbit for test chemicaland its structurally similar read across substances.. Based on the summarized studies for test chemical and its structurally and functionally similar read across substances, it can be concluded that the test chemical is unable to cause eye irritation and thus considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

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

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data of test substances

Justification for type of information:

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

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

WoE report is based on skin irritation studies as- WoE-2, WoE-3 and WoE-4.
The eye irritation studies were condcuted on rabbits to assess its skin irritating effects.

GLP compliance:

not specified

Species:

rabbit

Strain:

other: 1. and 2.New Zealand White 3.HC:NZW

Details on test animals or test system and environmental conditions:

1.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

2.TEST ANIMALS
- Weight at study initiation: 3 – 4 kg

3.- Sex: Female
- Age at study initiation: adult
- Weight at study initiation: 3.2-3.3 kg

Type of coverage:

other: 1.occlusive 2.not specified 3.semiocclusive

Preparation of test site:

other: 1.clipped 2.not specified 3.Mechanically depilated skin of flanks

Vehicle:

other: 1 and 2.unchanged (no vehicle) 3.water

Controls:

not specified

Amount / concentration applied:

1.500 mg (0.5g)
2.500 mg
3.500 mg

Duration of treatment / exposure:

1.4 hours
2.24 hours
3.4 hrs

Observation period:

1.60 min., 24, 48 and 72 hours after application.
2.7 days
3.1,24,48 and 72 hrs and day 7

Number of animals:

1.3 female rabbits
2. 2 rabbits
3.3 female rabbits

Details on study design:

1.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

2.TEST SITE
Area of exposure: Inner surface of ears
% coverage: Not mentioned
Type of wrap if used: Under an adhesive dressing

REMOVAL OF TEST SUBSTANCE
Washing (if done): The test substance was washed off with water and soap/vegetable oil.
Time after start of exposure: After the exposure period

SCORING SYSTEM: Not mentioned

3.SCORING SYSTEM: Draize method.

Irritation parameter:

other: 1.primary dermal irritation index (PDII)

Basis:

mean

Time point:

14 d

Score:

0

Max. score:

4

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritation parameter:

other: 2.overall irritation score

Basis:

mean

Time point:

7 d

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritation parameter:

other: 3.erythema and edema score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

4

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

1.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.
2.No skin irritation was observed.
3.None of the test animals produced skin reactions.

Other effects:

1.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.
2.No skin irritation was observed.
3.None of the test animals produced skin reactions.

1.

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

2.Not specified

Interpretation of results:

other: Not irritating

Conclusions:

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

Executive summary:

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

 

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. The animals were prepared 24 hours prior to application of test product. The furs from the dorsal area of trunk of animals were removed with electric clippers exposing an area measuring approximately 6 cm2 of body surface area of animal. The care was taken such that abrasion penetrated the Stratum corneum only and not dermis. 500gm (0.5g) of test compound was applied on a small area (approximately 6 cm2) of intact skin site. Each site of application was covered with impervious dressing which was secured in position with adhesive tape. 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 and the responses were scored according to Draize method. The Primary Irritation Index (PII) for Test chemical after 14 days of observation was 0.0.AlsoTest 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.

 

In next skin irritation study, approx. 500 mg of test chemical was applied to the inner surface of the ears of 2 rabbits. Male and Female New Zealand white rabbits were used for the study. The test chemical was applied under an adhesive dressing for 24 hours. At the end of the exposure period, the test substance was washed off with water and soap/vegetable oil. The observation period was 7 days. No skin irritation was observed after 7 days. Thus on the basis of the above study, the test substance, 7-amino-4-hydroxy-2-naphthalenesulfonic acid can be considered non-irritant to rabbit skin.

 

The above results were further supported by the Primary Skin irritation study conducted for test chemical to determine its skin irritating behavior in accordance with OECD guideline No. 404. The animals were subjected to a single 4- hour exposure to 500 mg of the test substance, mixed into a paste with water and applied semi-occlusively to the mechanically depilated skin of the flanks. The findings were scored after 1, 24, 48 and 72 hours as well as 7 days in accordance with the Draize method. The average irritation scores according to the Draize scoring for erythema and oedema were 0.0. Hence, the test chemical was considered to be not irritating to skin of female albino rabbits.

 

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

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

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

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data of test substances

Justification for type of information:

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

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

WoE report is based on eye irritation studies as- WoE-2, WoE-3 and WoE-4.
An eye irritation studies were condcuted on rabbits to assess its ocular effects.

GLP compliance:

not specified

Species:

rabbit

Strain:

other: 1. and 2.New Zealand White 2.not specified

Details on test animals or tissues and environmental conditions:

1.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,mad-libitum

2. Not specified

3.No data available

Vehicle:

other: 1 and 2.unchanged (no vehicle) 3.water

Controls:

yes

Amount / concentration applied:

1.100mg (0.1g)
2.50 mg
3.100 µl

Duration of treatment / exposure:

1.24 hours
2.24 hours
3.3 days

Observation period (in vivo):

1.The eyes were examined at 1, 24, 48 and 72 hours after test substance application
2.7 days
3.3 days

Duration of post- treatment incubation (in vitro):

no data available

Number of animals or in vitro replicates:

1.3 female rabbits
2.2 rabbits
3.6 rabbits

Details on study design:

1.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.

2.TEST SITE
Area of exposure: conjunctival sac
% coverage: no data
Type of wrap if used: no data

3.TEST SITE
- Area of exposure: the conjunctival sac of eye
- % coverage: No data
- Type of wrap if used: No data

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No data
- Time after start of exposure: No data
SCORING SYSTEM: No data

Irritation parameter:

other: 1.overall irritation score

Basis:

mean

Time point:

72 h

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritation parameter:

other: 2.overall irritation score

Basis:

mean

Time point:

7 d

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritation parameter:

other: 3.overall irritation score

Basis:

mean

Time point:

other: 3 days

Score:

0

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

1.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.
2.No eye irritation was observed.
3.Slight reddening in only one of 6 animals was observed but this was reversible after 3 days.

Other effects:

1.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.
2.No eye irritation was observed.
3.Slight reddening in only one of 6 animals was observed but this was reversible after 3 days.

1.

TABLE- 1     GRADING OF OCULAR LESIONS

 

S.NO/ SEX		OBSERVATION	Score				Total	Total Score
1/F			1 hr	24hrs	48 hrs	72 hrs
	Cornea	A.       Opacity-Degree of Density	0	0	0	0	0	0×0×5=0
		B.       Area of Cornea Involved	0	0	0	0	0
	Iris	A.       Values	0	0	0	0	0	0
	Conjunctivae	A.       Redness	1	0	0	0	1	0+0+0×5=5
		B.       Chemosis	0	0	0	0	0
		C.       Discharge	0	0	0	0	0
2/F	Cornea	A.       Opacity-Degree of Density	0	0	0	0	0	0×0×5=0
		B.       Area of Cornea Involved	0	0	0	0	0
	Iris	A.       Values	0	0	0	0	0	0
	Conjunctivae	A.       Redness	1	0	0	0	1	0+0+0×5=5
		B.       Chemosis	0	0	0	0	0
		C.       Discharge	0	0	0	0	0
3/F	Cornea	A.       Opacity-Degree of Density	0	0	0	0	0	0×0×5=0
		B.       Area of Cornea Involved	0	0	0	0	0
	Iris	A.       Values	0	0	0	0	0	0
	Conjunctivae	A.       Redness	1	0	0	0	1	0+0+0×5=5
		B.       Chemosis	0	0	0	0	0
		C.       Discharge	0	0	0	0	0
Grand total								0.00
Mean								0.00
Eye Irritation Scoring index								0.00

Interpretation of results:

other: not irritating

Conclusions:

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

Executive summary:

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

 

An ocular irritation study was conducted on New Zealand white rabbits in accordance with OECD 405 to assess the irritation parameter of Test chemical. Three female New Zealand White rabbits were used for the study. 0.1g of the undiluted test chemical was instilled in the conjunctival sac of rabbits after gently pulling the lower lid away from the eyeball. The other eye which remained untreated served as a control. The ocular lesions were evaluated at 1, 24, 48 and 72 hours after the treatment. The grades of ocular reactions (conjunctiva, cornea and iris) were recorded at each observation. To determine the reversibility of the effect the animals were observed normally for 21 days. Any other lesions in the eye viz pannus, staining were observed and scored accordingly. Examination of reactions was facilitated by use of biomicroscope and hand slit lamp. Individual animal weights before and during the study was observed. The overall irritation index of 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. The results obtained from these studies lead to a conclusion that Test chemical is indeed not irritating to eye. Hence, comparing the above annotations with the criteria of CLP regulation, Test chemical can be classified under the category “Not Classified”.

 

In next eye irritation study in rabbits, 50 mg of test chemical was instilled into the conjunctival sac of 2 animals. New Zealand white rabbits were used for the study. The animals were observed for 7 days. No eye irritation was observed after 7 days. Hence, the test chemical can be considered non-irritant to rabbit eyes.

 

The above results were further supported by the Eye irritation study conducted in 6 rabbits for test chemical to assess its eye irritation efficacy in accordance with OECD guideline 405. About 100 μl of chemical was placed into the conjunctival sac of each rabbits and the results were recorded for 3 days. Slight reddening in only one of 6 animals was observed but this was reversible after 3 days. Since the observed eye effects were not persisted after 3 days, the test chemical was considered to be not irritating to rabbits’ eye.

 

 

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

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the skin irritation potential 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 female New Zealand White rabbits. The animals were prepared 24 hours prior to application of test product. The furs from the dorsal area of trunk of animals were removed with electric clippers exposing an area measuring approximately 6 cm2 of body surface area of animal. The care was taken such that abrasion penetrated the Stratum corneum only and not dermis. 500gm (0.5g) of test compound was applied on a small area (approximately 6 cm2) of intact skin site. Each site of application was covered with impervious dressing which was secured in position with adhesive tape. 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 and the responses were scored according to Draize method. The Primary Irritation Index (PII) for Test chemical after 14 days of observation was 0.0.AlsoTest 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.

 

In next skin irritation study, approx. 500 mg of test chemical was applied to the inner surface of the ears of 2 rabbits. Male and Female New Zealand white rabbits were used for the study. The test chemical was applied under an adhesive dressing for 24 hours. At the end of the exposure period, the test substance was washed off with water and soap/vegetable oil. The observation period was 7 days. No skin irritation was observed after 7 days. Thus on the basis of the above study, the test substance, 7-amino-4-hydroxy-2-naphthalenesulfonic acid can be considered non-irritant to rabbit skin.

 

The above results were further supported by the Primary Skin irritation study conducted for test chemical to determine its skin irritating behavior in accordance with OECD guideline No. 404. The animals were subjected to a single 4- hour exposure to 500 mg of the test substance, mixed into a paste with water and applied semi-occlusively to the mechanically depilated skin of the flanks. The findings were scored after 1, 24, 48 and 72 hours as well as 7 days in accordance with the Draize method. The average irritation scores according to the Draize scoring for erythema and oedema were 0.0. Hence, the test chemical was considered to be not irritating to skin of female albino rabbits.

 

Based on the above summarized studies for test chemical and its structurally similar read across substances,it can be concluded that the testchemical is not able to cause skin irritation and considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

 

 

Eye irritation

Various studieshas been investigated for the test chemical to observe the potential for ocular irritation to a greater or lesser extent. The studies are based on in vivo experiments in rabbits for test chemicaland its structurally similar read across substanceswhich have beensummarized as below;

 

An ocular irritation study was conducted on New Zealand white rabbits in accordance with OECD 405 to assess the irritation parameter of Test chemical. Three female New Zealand White rabbits were used for the study. 0.1g of the undiluted test chemical was instilled in the conjunctival sac of rabbits after gently pulling the lower lid away from the eyeball. The other eye which remained untreated served as a control. The ocular lesions were evaluated at 1, 24, 48 and 72 hours after the treatment. The grades of ocular reactions (conjunctiva, cornea and iris) were recorded at each observation. To determine the reversibility of the effect the animals were observed normally for 21 days. Any other lesions in the eye viz pannus, staining were observed and scored accordingly. Examination of reactions was facilitated by use of biomicroscope and hand slit lamp. Individual animal weights before and during the study was observed. The overall irritation index of 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. The results obtained from these studies lead to a conclusion that test chemical is indeed not irritating to eye.

 

In next eye irritation study in rabbits, 50 mg of test chemical was instilled into the conjunctival sac of 2 animals. New Zealand white rabbits were used for the study. The animals were observed for 7 days. No eye irritation was observed after 7 days. Hence, the test chemical can be considered non-irritant to rabbit eyes.

 

The above results were further supported by the Eye irritation study conducted in 6 rabbits for test chemical to assess its eye irritation efficacy in accordance with OECD guideline 405. About 100 μl of chemical was placed into the conjunctival sac of each rabbits and the results were recorded for 3 days. Slight reddening in only one of 6 animals was observed but this was reversible after 3 days. Since the observed eye effects were not persisted after 3 days, the test chemical was considered to be not irritating to rabbits’ eye.

 

 

Based on the above summarized studies for test chemical and its structurally similar read across substances,it can be concluded that the testchemical is not able to cause eye irritation and considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

Justification for classification or non-classification

The skin and eye irritation potential of test chemical and its structurally and functionally similar read across substanceswere observed in various studies. The results obtained from these studies indicate that the chemical is unable to cause skin and eye irritation. Hence test chemical can be classified under the category “Not Classified” for skin and eye as per CLP.