2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4)

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 skin irritation potential of the test chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8). The studies are as mentioned below:

 

The skin irritation study of read across chemical was performed as per OECD Guidelines 404 in three female New Zealand White rabbits and complying with the GLP procedures. 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) fortest chemicalafter 14 days of observation was 0.0.Alsothe chemicaldid not produce pain and any clinical signs of toxicity throughout the examination period of 14 days. Hence, under the test conditions,the test substancecan be concluded to be not irritating to New Zealand White rabbit skin.

 

The above result was further supported by another experimental study conducted for similar read across chemical on rabbits’ ear to indicate its Comedogenicity and irritancy. The test chemical was mixed in propylene glycol at a 9 to 1 dilution for testing unless otherwise indicated (10% concentration). A colony of New Zealand albino rabbits that have genetically good ears and free from mites were used. Three rabbits, weighing two to three kilograms, were used for each assay. Animals were housed singly in suspended cages and fed Purina Rabbit Chow and water ad libitum. Animals were maintained on a 12-hour light and 12-hour dark cycle. A dose of 1 ml of the test material was applied and spread once daily to the entire inner surface of once for five days per week for two weeks. The opposite untreated ear of each animal served as an untreated control. The irritancy produced by repeated application of the chemical on the surface epidermis in the rabbit ear is evaluated on a scale of 0 to 5. The chemical falls under Grade 0 (no irritation observed). Hence it can be concluded that the test chemical was not irritating to rabbits’ skin.

 

Based on the above summarized studies for target chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8) 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

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the ocular irritation potential of the test chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8). The studies are as mentioned below:

 

The experimental study was conducted for the structurally similar read across substance in three female New Zealand White rabbits as per OECD Guidelines 405 and complying to the GLP procedures. About 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 did not produce any clinical signs of toxicity throughout the examination period of 21 days. Hence, under the test conditions, the chemical can be concluded to be not irritating to New Zealand White rabbit eyes.

 

The above result was further supported by an eye irritation study conducted by authoritative database for similar read across substance to investigate the local eye irritative potential of test chemical. Each rabbit received 2 installation of 0.2 ml of a 10% test chemical aqueous suspension into the conjunctival sac of rabbits 5 times per week for 4 weeks. The chemical produced only minimal irritating effects after 2 hours which could not confirmed the irritation potential. Hence, on the basis of findings the Test chemical can be considered as not irritating to the rabbits’ eye.

 

Based on the above summarized studies for target chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8) 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”.

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 read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

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 2 skin irritation studies as- WoE-2 and WoE-3. Skin irritation study of test chemical was conducted on rabbits to assess its skin irritating effects.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material: 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4)- Molecular formula: C26H25N5O19S6.4Na- Molecular weight: 995.85 g/mole- Smiles : [Na][Na][Na][Na]Nc1ccc2c(cc(S(O)(=O)=O)c(N=Nc3ccc(S(=O)(=O)CCOS(O)(=O)=O)cc3)c2O)c1N=Nc1ccc(S(=O)(=O)CCOS(O)(=O)=O) cc1S(O)(=O)=O- Substance type: Organic- Physical state: Solid powder

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

1.Details on test animalsAge: 10 to 12 weeksSex:FemaleBody weight range: 2.0kg±200gIdentification: By cage tag and corresponding colour body markingHousing: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, DelhiWater:Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles, ad libitumAcclimatization: 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.HUSBANDRYEnvironmental 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, IndiaWater : Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles,mad-libitum2.

Type of coverage:

other: 1.occlusive 2.Entire inner surface of one ear

Preparation of test site:

other: 1.clipped 2.A colony of rabbits that had genetically good ears and which were free from mites were used for the assay

Vehicle:

other: 1.unchanged (no vehicle) 2.The test chemical was mixed in propylene glycol at a 9 to 1 dilution for testing.

Controls:

not specified

Amount / concentration applied:

1.500 mg (0.5g)2.Dose of 1ml of the test material was applied once daily for five days per week for 2 weeks.

Duration of treatment / exposure:

1.4 hours2.once daily for five days per week for 2 weeks.

Observation period:

1.60 min., 24, 48 and 72 hours after application.2.2 weeks

Number of animals:

1.3 female rabbits2.3 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 waterOBSERVATION 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 SITEArea of exposure: Ear% coverage: A dose of 1ml of the test chemical is applied and spread once daily to the entire inner surface of one ear five days per week for 2 weeks.Type of wrap if used: No dataREMOVAL OF TEST SUBSTANCEWashing (if done): No dataTime after start of exposure: No dataSCORING SYSTEM: The irritancy produced by repeated application of a chemical or skin care product on the surface epidermis in the rabbit ear is evaluated on a scale of 0 to 5. The grades are summarized as follows: 0 = No irritation1 = few scales, no Erythema2 = diffuse scaling, no Erythema3 = Generalized scaling with Erythema4 = Scaling, Erythema and Edema5 = Epidermal necrosis and slough

Irritation parameter:

other: 1.overall irritation score

Basis:

mean

Time point:

14 d

Score:

0

Max. score:

8

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

other: 2.overall irritation score

Basis:

mean

Time point:

other: 2 weeks

Score:

0

Max. score:

5

Reversibility:

other: Not applicable

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 erythema observed

Other effects:

1.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. 2.No erythema observed

1.

TABLE

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.

Table : Ingredients and their Comedogenicity and Irritancy

Test chemical	Comedogenicity* [Grade 0-5]	Irritancy** [Grade 0-5]
test chemical	1	0

 

*Comedogenicity or the ability of test substance to produce follicular hyperkeratosis

** Irritancy or ability of test substance to produce surface epithelial irritation

 

Interpretation of results:

other: Not irritating

Conclusions:

The test material 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8) was considered to be not irritating to the skin of New Zealand White rabbits.

Executive summary:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the skin irritation potential of the test chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8). The studies are as mentioned below:

 

The skin irritation study of read across chemical was performed as per OECD Guidelines 404 in three female New Zealand White rabbits and complying with the GLP procedures. 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) fortest chemicalafter 14 days of observation was 0.0.Alsothe chemicaldid not produce pain and any clinical signs of toxicity throughout the examination period of 14 days. Hence, under the test conditions,the test substancecan be concluded to be not irritating to New Zealand White rabbit skin.

 

The above result was further supported by another experimental study conducted for similar read across chemical on rabbits’ ear to indicate its Comedogenicity and irritancy. The test chemical was mixed in propylene glycol at a 9 to 1 dilution for testing unless otherwise indicated (10% concentration). A colony of New Zealand albino rabbits that have genetically good ears and free from mites were used. Three rabbits, weighing two to three kilograms, were used for each assay. Animals were housed singly in suspended cages and fed Purina Rabbit Chow and water ad libitum. Animals were maintained on a 12-hour light and 12-hour dark cycle. A dose of 1 ml of the test material was applied and spread once daily to the entire inner surface of once for five days per week for two weeks. The opposite untreated ear of each animal served as an untreated control. The irritancy produced by repeated application of the chemical on the surface epidermis in the rabbit ear is evaluated on a scale of 0 to 5. The chemical falls under Grade 0 (no irritation observed). Hence it can be concluded that the test chemical was not irritating to rabbits’ skin.

 

Based on the above summarized studies for target chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8) 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”.

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 read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

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 2 eye irritation studies as- WoE-2 and WoE-3. An eye irritation study of test chemical was conducted on rabbits to assess its eye irritating effects.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material: 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4)- Molecular formula: C26H25N5O19S6.4Na- Molecular weight: 995.85 g/mole- Smiles : [Na][Na][Na][Na]Nc1ccc2c(cc(S(O)(=O)=O)c(N=Nc3ccc(S(=O)(=O)CCOS(O)(=O)=O)cc3)c2O)c1N=Nc1ccc(S(=O)(=O)CCOS(O)(=O)=O) cc1S(O)(=O)=O- Substance type: Organic- Physical state: Solid powder

Species:

rabbit

Strain:

other: 1.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.HUSBANDRYEnvironmental 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, IndiaWater : Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles,mad-libitum2.not specified

Vehicle:

other: 1.unchanged (no vehicle) 2.aqueous suspension

Controls:

yes

Amount / concentration applied:

1.100mg (0.1g)2.2 instillations of 10% (0.2ml)

Duration of treatment / exposure:

1.24 hours2.24 hours

Observation period (in vivo):

1.The eyes were examined at 1, 24, 48 and 72 hours after test substance application2.4 weeks

Duration of post- treatment incubation (in vitro):

No data available

Number of animals or in vitro replicates:

1.3 female 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 lampExamination 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.- Area of exposure: conjunctival sac of rabbits

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

110

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritation parameter:

other: 2. overall irritation score

Basis:

mean

Time point:

other: 4 weeks

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.Only minimal irritative effects after 2 hours.

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.2.2.Only minimal irritative effects after 2 hours.

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

2.Not specified

Interpretation of results:

other: not irritating

Conclusions:

The test material 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8) was considered to be not irritating to the rabbits' eye.

Executive summary:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the ocular irritation potential of the test chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8). The studies are as mentioned below:

 

The experimental study was conducted for the structurally similar read across substance in three female New Zealand White rabbits as per OECD Guidelines 405 and complying to the GLP procedures. About 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 did not produce any clinical signs of toxicity throughout the examination period of 21 days. Hence, under the test conditions, the chemical can be concluded to be not irritating to New Zealand White rabbit eyes.

 

The above result was further supported by an eye irritation study conducted by authoritative database for similar read across substance to investigate the local eye irritative potential of test chemical. Each rabbit received 2 installation of 0.2 ml of a 10% test chemical aqueous suspension into the conjunctival sac of rabbits 5 times per week for 4 weeks. The chemical produced only minimal irritating effects after 2 hours which could not confirmed the irritation potential. Hence, on the basis of findings the Test chemical can be considered as not irritating to the rabbits’ eye.

 

Based on the above summarized studies for target chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8) 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”.

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 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8). The studies are as mentioned below:

 

The skin irritation study of read across chemical was performed as per OECD Guidelines 404 in three female New Zealand White rabbits and complying with the GLP procedures. 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) fortest chemicalafter 14 days of observation was 0.0.Alsothe chemicaldid not produce pain and any clinical signs of toxicity throughout the examination period of 14 days. Hence, under the test conditions,the test substancecan be concluded to be not irritating to New Zealand White rabbit skin.

 

The above result was further supported by another experimental study conducted for similar read across chemical on rabbits’ ear to indicate its Comedogenicity and irritancy. The test chemical was mixed in propylene glycol at a 9 to 1 dilution for testing unless otherwise indicated (10% concentration). A colony of New Zealand albino rabbits that have genetically good ears and free from mites were used. Three rabbits, weighing two to three kilograms, were used for each assay. Animals were housed singly in suspended cages and fed Purina Rabbit Chow and water ad libitum. Animals were maintained on a 12-hour light and 12-hour dark cycle. A dose of 1 ml of the test material was applied and spread once daily to the entire inner surface of once for five days per week for two weeks. The opposite untreated ear of each animal served as an untreated control. The irritancy produced by repeated application of the chemical on the surface epidermis in the rabbit ear is evaluated on a scale of 0 to 5. The chemical falls under Grade 0 (no irritation observed). Hence it can be concluded that the test chemical was not irritating to rabbits’ skin.

 

Based on the above summarized studies for target chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8) 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

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the ocular irritation potential of the test chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8). The studies are as mentioned below:

 

The experimental study was conducted for the structurally similar read across substance in three female New Zealand White rabbits as per OECD Guidelines 405 and complying to the GLP procedures. About 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 did not produce any clinical signs of toxicity throughout the examination period of 21 days. Hence, under the test conditions, the chemical can be concluded to be not irritating to New Zealand White rabbit eyes.

 

The above result was further supported by an eye irritation study conducted by authoritative database for similar read across substance to investigate the local eye irritative potential of test chemical. Each rabbit received 2 installation of 0.2 ml of a 10% test chemical aqueous suspension into the conjunctival sac of rabbits 5 times per week for 4 weeks. The chemical produced only minimal irritating effects after 2 hours which could not confirmed the irritation potential. Hence, on the basis of findings the Test chemical can be considered as not irritating to the rabbits’ eye.

 

Based on the above summarized studies for target chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8) 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 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8)  and its structurally and functionally similar read across substanceswere observed in various studies. The results obtained from these studies indicate that the chemical  is unlikely to cause skin and eye irritation. Hence 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (CAS No: 607724-37-8) can be classified under the category “Not Classified” for skin and eye as per CLP.