Reaction products of diazotised sodium 2-amino-5-{[2-(sulfonatooxy)ethyl]sulfonyl}benzenesulfonate coupled with sodium 2-(4-aminobenzene-1-sulfonyl)ethyl sulfate, subsequently coupled with diazotised sodium 2-(4-aminobenzene-1-sulfonyl)ethyl sulfate

Endpoint summary

Description of key information

Skin irritation:

1. The intracutenous injection did not induce any cutenous damage to the skin. A black spot formed on the on back skin disappeared largely during the second day and was completely invisible after 2 days. Hence, the test chemical can be considered to be not irritating to skin.

2. Hence, it was concluded that Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested and classified as “Category- Not Classified” as per CLP Classification.

 

Eye irritation:

1. Slight erythema was observed in the rabbit eyes which disappeared in 24 hours. No corneal damage was observed. Hence, the test chemical can be considered to be not irritating to eyes.

2. The eye potential of the test chemical was evaluated in rabbits. Following application of the neat test material, all animals showed slight conjunctival redness which in five of 6 animals was completely reversible within 72 hours, and in one animal within 7 days. In two animals, very slight tear flow occurred after application which also was completely reversible within 48 hours.
Following application of a 30% aqueous solution of the test material, all animals showed slight conjunctival redness (max. score 1) one hour after application which was completely reversible within 24 hours. Following application of a 3% aqueous solution, three animals showed slight conjunctival redness (max. score 1) at 24 hours after application which was completely reversible within 48 hours. Since the observed effects were reversible by 7 days, the test chemical can be considered to be not irritating to rabbit eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Remarks:

Read across data

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 Irritation/corrosion potential of Tetra sodium 1-acetamido-2-hydroxy-3- (4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate in Sprague Dawley rats

GLP compliance:

yes

Specific details on test material used for the study:

IUPAC Name: Tetrasodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate
Common Name: C.I. Food Black 1, Brilliant Black 1
Molecular Formula:C28H21N5O14S4.4Na
Molecular Weight: 867.6873 g/mol
Substance Type: Organic
Physical state: Solid

SOURCE OF TEST MATERIAL
- Test Item: Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate (CAS No. 2519-30-4)
- Source of test material: Sustainability Support Services (Europe) AB
- Batch No.of test material: FG/15-16/1324
- Manufacturing Date: Aug., 2015
- Expiration date of the lot/batch: July, 2023
- Purity test date: No data
- Consistency: Solid, powder
- Colour: Black

RADIOLABELLING INFORMATION (not applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient Temperature
- Stability under test conditions: No data
- Solubility and stability of the test substance in the solvent/vehicle: No data
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: No data

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test item was moistened with distilled water before application.
- Preliminary purification step (if any):No data
- Final dilution of a dissolved solid, stock liquid or gel: No data
- Final preparation of a solid: No data

FORM AS APPLIED IN THE TEST (if different from that of starting material): Paste

OTHER SPECIFICS:
Safety Precautions : Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).

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:

water

Remarks:

(Distilled water)

Controls:

not specified

Amount / concentration applied:

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

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

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

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

Duration of treatment / exposure:

24 hrs.

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 total 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:

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.

Laboratory Test Item Code :TAS/122/013

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	0 - 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	0 - 14	0/5

 

 

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

 

ndividual Animal -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	0 - 14	0
				2	0 - 14	0
				3	0 - 14	0
				4	0 - 14	0
				5	0 - 14	0

 

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	0 - 14	0
				7	0 - 14	0
				8	0 - 14	0
				9	0 - 14	0
				10	0 - 14	0

 

Individual Animal - Body Weight and Percent Body Weight Gain (g)

Test System : Sprague Dawley Rat

Sex    : Male

Group : I

Dose  : 2000 mg/kg body weight

 

Animal No.	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
1	266.4	292.3	9.72	305.0	4.34	14.49
2	269.2	288.6	7.21	310.3	7.52	15.27
3	270.3	296.0	9.51	315.0	6.42	16.54
4	271.3	299.4	10.36	319.6	6.75	17.80
5	272.5	304.2	11.63	322.7	6.08	18.42

Sex    : Female

Group : I

Dose  : 2000 mg/kg body weight

 

Animal No.	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
6	239.1	255.3	6.78	267.3	4.70	11.79
7	242.4	250.7	3.42	258.3	3.03	6.56
8	243.5	253.8	4.23	262.4	3.39	7.76
9	247.4	260.4	5.25	269.8	3.61	9.05
10	250.4	264.0	5.43	276.5	4.73	10.42

 

Individual Animal - Gross Pathological Findings

Test System : Sprague Dawley Rat

Sex    : Male

Group : I

Dose  : 2000 mg/kg body weight

 

Animal No.	Fate	Gross Pathological Findings
1	TS	No abnormality detected
2	TS	No abnormality detected
3	TS	No abnormality detected
4	TS	No abnormality detected
5	TS	No abnormality detected

 

Sex    : Female

Group : I

Dose  : 2000 mg/kg body weight

 

Animal No.	Fate	Gross Pathological Findings
6	TS	No abnormality detected
7	TS	No abnormality detected
8	TS	No abnormality detected
9	TS	No abnormality detected
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 found at the end of 14 days observation period after patch removal.
Hence, it was concluded that Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested and classified as “Category- Not Classified” as per CLP Classification.

Executive summary:

A study was designed and conducted to determine the dermal reaction profile of the read across substance Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate[CAS:2519 -30 -4] 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 moistened with distilled water. The test item was applied 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 Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate 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. 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. The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) were found at the end of 14 days observation period after patch removal. Hence, it was concluded that Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested and classified as “Category- Not Classified” as per CLP Classification.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Remarks:

Read across data

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data is from safety assessment reports

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

To determine the skin irritation potential of the test chemical in rabbits

GLP compliance:

not specified

Species:

rabbit

Strain:

not specified

Details on test animals or test system and environmental conditions:

no data available

Type of coverage:

not specified

Preparation of test site:

not specified

Vehicle:

not specified

Controls:

not specified

Amount / concentration applied:

0.1 ml 5% solution

Duration of treatment / exposure:

no data available

Observation period:

no data available

Number of animals:

no data available

Details on study design:

TEST SITE
- Area of exposure: back skin of rabbits

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

48 h

Reversibility:

fully reversible within: 2 days

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No cutaneous damage was observed. A black spot formed on the on back skin disappeared largely during the second day and was completely invisible after 2 days.

Other effects:

no data available

Interpretation of results:

other: not irritating

Conclusions:

The intracutenous injection did not induce any cutenous damage to the skin. A black spot formed on the on back skin disappeared largely during the second day and was completely invisible after 2 days. Hence, the test chemical can be considered to be not irritating to skin.

Executive summary:

The skin irritation potential of the read across substance Tetrasodium (3E)-6 -amino-4 -oxo-3 -(2 -{7 -sulfinato-4 -[(E)-2 -(4 -sulfonatophenyl)diazen-1 -yl]naphthalen-1 -yl}hydrazin-1 -ylidene)-3,4 -dihydronaphthalene-2,7 -disulfonate (CAS no.: 2118 -39 -0, E.C. no.: 218 -326 -9) was assessed in rabbits. Rabbits were given intracutenous injection of 0.1 ml 5% solution of the test chemical on their back skin and observed for signs of dermal reactions (exact duration of study not mentioned). The intracutenous injection did not induce any cutenous damage to the skin. A black spot formed on the on back skin disappeared largely during the second day and was completely invisible after 2 days.Hence, the test chemical can be considered to be not irritating to skin, as per the CLP criteria .

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Remarks:

Read across data

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Justification for type of information:

data is from experimental reports of read across subtance

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

To assess the eye irritation potential of the test chemical in rabbits

GLP compliance:

not specified

Species:

rabbit

Strain:

not specified

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Hacking & Churchill, Ltd., UK)
- Age at study initiation: no data available
- Weight at study initiation: no data available
- Housing:The animals were conventionally kept in single cages
- Diet (e.g. ad libitum): appropriate diet, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: no data available

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

About 100 mg of the finely powdered test specimen and 100 microliters of a 30% or 3% strength aqueous solution of the test chemical

Duration of treatment / exposure:

single exposure

Observation period (in vivo):

The findings were obtained 1, 24, HS and 72 hours after administration and at the end of the follow-up period of 7 days

Duration of post- treatment incubation (in vitro):

no data available

Number of animals or in vitro replicates:

6

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no
- Time after start of exposure: no

SCORING SYSTEM: The condition of cornea, iris and conjunctivae according to DRAIZE was assessed. The condition of cornea, iris and conjunctivae according to DRAIZE was assessed (see rating table). All other findings not covered by the DRAIZE scale were also recorded. For evaluation, the DRAIZE grades were used at approximately 24, 48, and 72 hours after application.

TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein : no data available

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

fully reversible within: 7 days

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

fully reversible within: 7 days

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

24/48/72 h

Score:

1

Reversibility:

fully reversible within: 7 days

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Following application of the neat test material, all animals showed slight conjunctival redness which in five of 6 animals was completely reversible within 72 hours, and in one animal within 7 days.
In two animals, very slight tear flow occurred after application which also was completely reversible within 48 hours.
Following application of a 30% aqueous solution of the test material, all animals showed slight conjunctival redness (max. score 1) one hour after application which was completely reversible within 24 hours.
Following application of a 3% aqueous solution, three animals showed slight conjunctival redness (max. score 1) at 24 hours after application which was completely reversible within 48 hours.

Table #: Irritant/corrosive response data for neat substance  for each animal at each observation time up to removal of each animal from the test

Score at time point / Reversibility	Cornea	Iris	Conjunctivae	Chemosis
	Max. score: 4	Max. score: 2	Max. score: 3	Max. score: 4
60 min	0/0/0/0/0/0	0/0/0/0/0/0	1/1/2/1/1/1	0/2/1/0/1/0
24 h	0/0/0/0/0/0	0/0/0/0/0/0	1/2/1/1/0/2	0/0/0/0/0/0
48 h	0/0/0/0/0/0	0/0/0/0/0/0	1/1/1/1/0/1	0/0/0/0/0/0
72 h	0/0/0/0/0/0	0/0/0/0/0/0	0/1/0/0/0/0	0/0/0/0/0/0
7 d	0/0/0/0/0/0	0/0/0/0/0/0	0/0/0/0/0/0	0/0/0/0/0/0
Average 24h, 48h, 72h	0/0/0/0/0/0	0/0/0/0/0/0	0.6/1.3/0.6/0.6/0.0/1.0	0/0/0/0/0/0
Area effected	0/0/0/0/0/0
Reversibility*)			c.	c.
Average time (unit) for reversion			7 d	24 h

*) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

Judgment is based on mean scores for each animal following grading at 24, 48 and 72 hours after application of the test material, in consideration of reversibility.

Interpretation of results:

other: not irritating

Conclusions:

The eye potential of the test chemical was evaluated in rabbits. Following application of the neat test material, all animals showed slight conjunctival redness which in five of 6 animals was completely reversible within 72 hours, and in one animal within 7 days. In two animals, very slight tear flow occurred after application which also was completely reversible within 48 hours.
Following application of a 30% aqueous solution of the test material, all animals showed slight conjunctival redness (max. score 1) one hour after application which was completely reversible within 24 hours. Following application of a 3% aqueous solution, three animals showed slight conjunctival redness (max. score 1) at 24 hours after application which was completely reversible within 48 hours. Since the observed effects were reversible by 7 days, the test chemical can be considered to be not irritating to rabbit eyes.

Executive summary:

The eye potential of the read across substance Trisodium 8-hydroxy-1,3,6-pyrenetrisulfonate (CAS no.: 6358-69-6) was evaluated in rabbits. 6 albino rabbits were used for the study. The animals were conventionally kept in single cages and were given an appropriate diet and tap water ad libitum. For each of the 6 rabbits, about 100 mg of the finely powdered test specimen and 100 microliters of a 30% or 3% strength aqueous test chemical solution were applied to the conjunctival sac of one eye. Only animals with intact eyes were used.The untreated eye served as control. The findings were obtained 1, 24, 48 and 72 hours after administration and at the end of the follow-up period of 7 days. The condition of cornea, iris and conjunctivae according to DRAIZE was assessed. Following application of the neat test material, all animals showed slight conjunctival redness which in five of 6 animals was completely reversible within 72 hours, and in one animal within 7 days. In two animals, very slight tear flow occurred after application which also was completely reversible within 48 hours. Following application of a 30% aqueous solution of the test material, all animals showed slight conjunctival redness (max. score 1) one hour after application which was completely reversible within 24 hours.  Following application of a 3% aqueous solution, three animals showed slight conjunctival redness (max. score 1) at 24 hours after application which was completely reversible within 48 hours. Since the observed effects were reversible by 7 days, the test chemical can be considered to be not irritating to rabbit eyes.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Remarks:

read across data

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

data is from safety assessment reports

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

To determine the eye irritation potential of the test chemical in rabbits

GLP compliance:

not specified

Species:

rabbit

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Not specified

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

50 mg

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

24 hours

Duration of post- treatment incubation (in vitro):

no data available

Number of animals or in vitro replicates:

3

Details on study design:

TEST SITE
- Area of exposure: conjunctival sac

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24 h

Reversibility:

fully reversible within: 24 hours

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Slight erythema was observed in the rabbit eyes which disappeared in 24 hours. No corneal damage was observed.

Interpretation of results:

other: not irritating

Conclusions:

Slight erythema was observed in the rabbit eyes which disappeared in 24 hours. No corneal damage was observed. Hence, the test chemical can be considered to be not irritating to eyes.

Executive summary:

The eye irritation potential of the read across substance Tetrasodium (3E)-6 -amino-4 -oxo-3 -(2 -{7 -sulfinato-4 -[(E)-2 -(4 -sulfonatophenyl)diazen-1 -yl]naphthalen-1 -yl}hydrazin-1 -ylidene)-3,4 -dihydronaphthalene-2,7 -disulfonate (CAS no.: 2118 -39 -0, E.C. no.: 218 -326 -9) in rabbits. The undiluted test substance was instilled in conjuctival sac of 2 rabbits at a concentration of 50mg. There was slight erythema observed in the rabbit eyes which disappeared in 24 hours. No corneal damage was observed in any of the animals. Hence, the test chemical can be considered to be not irritating to eyes of rabbits.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

1. The skin irritation potential of the read across substance Tetrasodium (3E)-6 -amino-4 -oxo-3 -(2 -{7 -sulfinato-4 -[(E)-2 -(4 -sulfonatophenyl)diazen-1 -yl]naphthalen-1 -yl}hydrazin-1 -ylidene)-3,4 -dihydronaphthalene-2,7 -disulfonate (CAS no.: 2118 -39 -0, E.C. no.: 218 -326 -9) was assessed in rabbits. Rabbits were given intracutenous injection of 0.1 ml 5% solution of the test chemical on their back skin and observed for signs of dermal reactions (exact duration of study not mentioned). The intracutenous injection did not induce any cutenous damage to the skin. A black spot formed on the on back skin disappeared largely during the second day and was completely invisible after 2 days.Hence, the test chemical can be considered to be not irritating to skin, as per the CLP criteria .

 

2. A study was designed and conducted to determine the dermal reaction profile of the read across substance Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate[CAS:2519 -30 -4] 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 moistened with distilled water. The test item was applied 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 Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate 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. 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. The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) were found at the end of 14 days observation period after patch removal. Hence, it was concluded that Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested and classified as “Category- Not Classified” as per CLP Classification.

 

Eye irritation:

1. The eye irritation potential of the read across substance Tetrasodium (3E)-6 -amino-4 -oxo-3 -(2 -{7 -sulfinato-4 -[(E)-2 -(4 -sulfonatophenyl)diazen-1 -yl]naphthalen-1 -yl}hydrazin-1 -ylidene)-3,4 -dihydronaphthalene-2,7 -disulfonate (CAS no.: 2118 -39 -0, E.C. no.: 218 -326 -9) in rabbits. The undiluted test substance was instilled in conjuctival sac of 2 rabbits at a concentration of 50mg. There was slight erythema observed in the rabbit eyes which disappeared in 24 hours. No corneal damage was observed in any of the animals. Hence, the test chemical can be considered to be not irritating to eyes of rabbits.

 

2. The eye potential of the read across substance Trisodium 8-hydroxy-1,3,6-pyrenetrisulfonate (CAS no.: 6358-69-6) was evaluated in rabbits. 6 albino rabbits were used for the study. The animals were conventionally kept in single cages and were given an appropriate diet and tap water ad libitum. For each of the 6 rabbits, about 100 mg of the finely powdered test specimen and 100 microliters of a 30% or 3% strength aqueous test chemical solution were applied to the conjunctival sac of one eye. Only animals with intact eyes were used.The untreated eye served as control. The findings were obtained 1, 24, 48 and 72 hours after administration and at the end of the follow-up period of 7 days. The condition of cornea, iris and conjunctivae according to DRAIZE was assessed. Following application of the neat test material, all animals showed slight conjunctival redness which in five of 6 animals was completely reversible within 72 hours, and in one animal within 7 days. In two animals, very slight tear flow occurred after application which also was completely reversible within 48 hours. Following application of a 30% aqueous solution of the test material, all animals showed slight conjunctival redness (max. score 1) one hour after application which was completely reversible within 24 hours.  Following application of a 3% aqueous solution, three animals showed slight conjunctival redness (max. score 1) at 24 hours after application which was completely reversible within 48 hours. Since the observed effects were reversible by 7 days, the test chemical can be considered to be not irritating to rabbit eyes.

 

 

Justification for classification or non-classification

Based on the available data and weight of evidence approach, the test substance did not show any reaction to the skin and eyes of the animals. Hence, the test substance was considered to be not irritating to the skin and eyes.