2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated

Endpoint summary

Description of key information

Acute oral toxicity:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the acute oral toxicity of the test chemical 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3).The studies are as mentioned below:

1.Acute oral toxicity study of test chemical was conducted in 40 Crj: CD (SD) IGS, SPF male and female rat at the concentration of 0, 500, 1000 and 2000 mg/kg bw. The test substance (Purity - >99 %; Obtained from - Dainippon Ink & Chemicals, Inc. and lot number - 000207) was suspended in a medium containing 0.1% Tween 80 added 0.5% CMC-Na aqueous solution as 20 mL/kg. Mortality and general condition were observed over 4 days, 30 minutes, 1, 3 and 6 hours after administration, once a day for 14 days, thereafter. Body weight was measured using an epple dish balance on days 4, 8 and 15 immediately before administration.Animals were observed for clinical signs. No death occurred in both males and females.Clinical signs observed such as, red feces exhibiting the same color tone as the test substance were found between 6 hours and 3 days after administration in males and females of the test substance - administered group, and in all the sexes on the 2nd day. In addition, the coloration of the coat considered to originate from this red flavor was observed in males and females of the test substance-administered group between the 2nd and 4th days. But no abnormality considered to be a toxicity change was observed. No abnormality was found in body weight of animals. Diaphragmatic hernia in the thoracic cavity was found in one female in the 1000 mg/kg group. In the hernia, the caudate portion of the liver protruded nodularly into the thoracic cavity, a part of which was adhered to the chest wall. Since this change was expressed only in one case and not related to the dose, it was judged as a contingent finding. No other abnormality was found. Therefore,LD50 was considered to be >2000 mg/kg bw, when Male and female SD rats were treated with test chemical via oral route.

2.Acute oral toxicity study was done infemale Sprague Dawley rats using test chemical.Distilled water was used as vehicle.Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Step - II). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality after the dosing. No mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - I). Administration of the test item at 2000 mg/kg resulted in diarrhoea (black colour stools) in all animals with onset at 2 hours and no mortality after the dosing. As no mortality were observed at 24 hours after the dosing, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - II). Administration of the test item at 2000 mg/kg resulted in diarrhoea (black colour stools) in all animals with onset at 4 hours and no mortality after the dosing. All animals from 300 mg/kg and 2000 mg/kg dose groups survived through the study period of 14 days. Staining of the stool is attributed to the black colour of the test item. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.Hence,LD50 value was considered to be5000 mg/kg bw,when female Sprague Dawleyrats were treated with test chemical orally via gavage following 14 days of observation period according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

3.In acute oral toxicity study, mice were treated with test chemical orally. 50 % Mortality was observed in treated mice at 20,000 mg/kg bw. Therefore, LD50 was considered to be 20,000 mg/kg bw. When mice were treated with test chemical orally.

Thus, based on the above summarised studies,2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3)and it’s structurallyand functionally similarread across substance, it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation,2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3)cannot be classified for acute oral toxicity.Hence,based on the data available for the structurally and functionally similar read across, test chemical 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3) is not likely to be toxic atleast at the dose of >2000 mg/Kg bw.

Acute inhalation toxicity:

2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated has very low  vapor pressure (2.0265E-17 Pa), so the potential for the generation of inhalable vapours is very low. Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point for repeated inhalation toxicity was considered for waiver.

Acute dermal toxicity:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine theAcute dermal toxicityof the test chemical 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3).The studies are as mentioned below:

1.In acute dermal toxicity study,male and female Sprague Dawley rats were treated with test chemical in the concentration of 2000 mg/kg bw by dermal application.Distilled water was used as vehicle. The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.Hence,The LD50 value was considered to be >2000 mg/kg bw,when male and female Sprague Dawley rats were semiocclusively treated with test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

2.In an acute dermal toxicity study, Wistar male and female rats were treated with test chemical in the concentration of 2000 mg/kg bw by dermal application. No mortality and any clinical sign of toxicity were observed throughout the observation period of 14 days in treated rats. The body weight of each animal recorded on day 0, 7th and 14th showed normal increase and there was no significant increase or decrease in weight was recorded. Therefore, LD50 was considered to be > 2000 mg/kg bw,when Wistar male and female rats were treated with 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs by dermal application.

3.In acute dermal toxicity study,rabbits were treated with test chemical in the concentration of 10000 mg/kg bw by dermal application.No mortality was observed in treated rabbits at dose 10000 mg/kg bw.Therefore, LD50 value was considered to be >10000 mg/kg bw,when rabbits were treated with test chemical  by dermal application.

Thus, based on the above summarised studies,2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3)and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3)cannot be classified for acute dermal toxicity.Hence,based on the data available for the structurally and functionally similar read across chemical, 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3) is not likely to be toxic atleast at the dose of >10000 mg/Kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

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

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 two acute oral toxicity studies as- 1., 2. and 3. Acute Oral toxicity test was carried out to study the effects of the test chemicals on rodents

GLP compliance:

not specified

Test type:

other: 1.no data available 2.acute toxic class method 3.no data available

Limit test:

yes

Specific details on test material used for the study:

2.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 (if applicable)- Radiochemical purity: No data- Specific activity: No data- Locations of the label: No data- Expiration date of radiochemical substance: No dataSTABILITY AND STORAGE CONDITIONS OF TEST MATERIAL- Storage condition of test material: Test Item was stored at 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 dataTREATMENT OF TEST MATERIAL PRIOR TO TESTING- Treatment of test material prior to testing: Test item was suspended in distilled water. The formulation was prepared fresh on the day of dosing. - 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 dataFORM AS APPLIED IN THE TEST (if different from that of starting material) : No dataOTHER SPECIFICS: Safety Precautions: Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).

Species:

other: 1.rat 2.rat 3.mouse

Strain:

other: 1.Crj: CD (SD) IGS, SPF 2.Sprague-Dawley 3.not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

1.Details on test animalTEST ANIMALS- Source: Charles River Japan Co., Ltd.- Age at study initiation: 5 weeks old- Weight at study initiation: The body weight ranged from 116 to 127 g for males and 106 to 124 g for females.- Fasting period before study: Rats fasted for about 17 hours from the day before administration.- Housing: 5 animals (same sex) were housed and raised in polycarbonate cages spread with experimental animal bedding.- Diet (e.g. ad libitum): solid feed- Water (e.g. ad libitum): Freely ingested tap water irradiated with ultraviolet rays after filter filtration with a pore size of 5 μm.- Acclimation period: 5 daysENVIRONMENTAL CONDITIONS- Temperature (°C): 22 ± 2 ° C- Humidity (%): 55 ± 15%- Air changes (per hr): ventilation at about 12times / hour- Photoperiod (hrs dark / hrs light): lighting 12 hours / day (7: 00-19: 00).2.TEST ANIMALS- Source: National Institute of Biosciences, Pune.- Females nulliparous and non-pregnant: yes- Age at study initiation: Female rats of the age of approximately 8 to 12 weeks old were used.- Weight at study initiation: Body weight range was 199.1 to 219.9 grams.Body weights at the start : FemaleMean : 206.81 g (= 100 %)Minimum : 199.1 g (- 3.73 %)Maximum : 219.9 g (+ 6.33 %) Total No. of animals : 12- Fasting period before study: Approximately 16 hours or more.- Housing: The rats were housed in polycarbonate cages.- 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): 20.6 to 23.2 degree centigrade.- Humidity (%): 55.1% to 58.6%.- 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: 26-09-2016 to 15-10-2016

Route of administration:

other: 1.oral: gavage 2.oral: gavage 3.oral: unspecified

Vehicle:

other: 1.CMC (carboxymethyl cellulose) 2.Distilled water 3.not specified

Details on oral exposure:

1.Details on exposureVEHICLE- Amount of vehicle (if gavage): 20 mL / kg- DOSAGE PREPARATION (if unusual): The test substance was suspended in a medium containing 0.1% Tween 80 added 0.5% CMC-Na aqueous solution.2.VEHICLE- Concentration in vehicle: 300 mg/kg, 300 mg/kg, 2000 mg/kg and 2000 mg/kg- Amount of vehicle (if gavage): No data- Justification for choice of vehicle: No data- Lot/batch no. (if required): No data- Purity: No dataMAXIMUM DOSE VOLUME APPLIED: 10 ml per kg of body weight.DOSAGE PREPARATION (if unusual): No dataCLASS METHOD (if applicable)- Rationale for the selection of the starting dose: No data3.No data available

Doses:

1.0, 500, 1000 and 2000 mg/kg2.Dose Group I : 300 mg/kgDose Group I : 300 mg/kgDose Group II : 2000 mg/kgDose Group II : 2000 mg/kg3.20000 mg/kg bw

No. of animals per sex per dose:

1.Total = 30 (sex/dose)2.Three females were used at each step. 3.No data available

Control animals:

other: 1.Total = 10 (Male/Female) 2.not specified 3.No data available

Details on study design:

1.Details on study design- Duration of observation period following administration: 14 days - Frequency of observations: Mortality and general condition were observed over 4 days, 30 minutes, 1, 3 and 6 hours afteradministration, once a day for 14 days, thereafter; and weighing: Body weight was measured using an epple dish balance on days 4, 8 and 15 immediately beforeadministration.- Necropsy of survivors performed: yes- Other examinations performed: Animals were observed for clinical signs. 2.- Duration of observation period following administration: 14 days - Frequency of observations and weighing: Twice daily- Necropsy of survivors performed: yes- Other examinations performed: Clinical Observations and General Appearance:Animals were observed for clinical signs, mortality and morbidity, until sacrifice.Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at immediately (0 to 5 minutes), 5, 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily therea fter for 14 day. Daily observation was done as far as possible at the same time.Body weights:Individual animal body weights were recorded, before fasting, prior to administration of the test item (fasting body weights), weekly thereafter and at termination on day 14. Weight changes were calculated and recorded.Gross Pathology:Necropsy was performed on all animals at the end of the study period on day 15. 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. Histopathology:No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.3.No data available

Statistics:

No data available

Preliminary study:

No data available

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality was observed at 2000 mg/kg bw.

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality was observed

Sex:

not specified

Dose descriptor:

LD50

Effect level:

20 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: 50% Mortality was observed in treated mice

Mortality:

1.No mortality was observed at 2000 mg/kg bw.2.All animals treated at the dose level of 300 mg/kg body weight and 2000mg/kg body weight survived through the study period of 14 days.3.50 % Mortality was observed in treated mice

Clinical signs:

1.Clinical signs such as, red feces exhibiting the same color tone as the test substance were found between 6 hours and 3 days after administration in males and females of the test substance - administered group, and in all the sexes on the 2nd day. In addition, the coloration of the coat considered to originate from this red flavor was observed in males and females of the test substance-administered group between the 2nd and 4th days. But no abnormality considered to be a toxicity change was observed. 2.Group I Step I :Animals treated at the dose level of 300 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days.Group I Step II :Animals treated at the dose level of 300 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days.Group II Step I :Animals treated at the dose level of 2000 mg/kg body weight resulted in diarrhoea (black colour stools) in all animals with onset at 2 hours after the dosing. All animals survived through the study period of 14 days and were free of signs of toxicity on day 1 after the dosing.Group II Step II :Animals treated at the dose level of 2000 mg/kg body weight resulted in diarrhoea (black colour stools) in all animals with onset at 4 hours after the dosing. All animals survived through the study period of 14 days and were free of signs of toxicity on day 1 after the dosing.Staining of the stool is attributed to the black colour of the test item.3.No data available

Body weight:

1.No abnormality was found in body weight of animals.2.Group I Step I (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.45% and 12.76% respectively. Group I Step II (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 3.56% and 12.49% respectively. Group II Step I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 3.06% and 10.98% respectively. Group II Step II (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 2.17% and 9.86% respectively. 3.No data available

Gross pathology:

1.Diaphragmatic hernia in the thoracic cavity was found in one female in the 1000 mg/kg group. In the hernia, the caudate portion of the liver protruded nodularly into the thoracic cavity, a part of which was adhered to the chest wall. Since this change was expressed only in one case and not related to the dose, it was judged as a contingent finding. No other abnormality was found.2.Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.3.No data available

Other findings:

No data available

Interpretation of results:

other: not classified

Conclusions:

The test chemical 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3) is not likely to be toxic atleast at the dose of >20000 mg/kg bw.

Executive summary:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the acute oral toxicity of the test chemical 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3).The studies are as mentioned below:

1.Acute oral toxicity study of test chemical was conducted in 40 Crj: CD (SD) IGS, SPF male and female rat at the concentration of 0, 500, 1000 and 2000 mg/kg bw. The test substance (Purity - >99 %; Obtained from - Dainippon Ink & Chemicals, Inc. and lot number - 000207) was suspended in a medium containing 0.1% Tween 80 added 0.5% CMC-Na aqueous solution as 20 mL/kg. Mortality and general condition were observed over 4 days, 30 minutes, 1, 3 and 6 hours after administration, once a day for 14 days, thereafter. Body weight was measured using an epple dish balance on days 4, 8 and 15 immediately before administration.Animals were observed for clinical signs. No death occurred in both males and females.Clinical signs observed such as, red feces exhibiting the same color tone as the test substance were found between 6 hours and 3 days after administration in males and females of the test substance - administered group, and in all the sexes on the 2nd day. In addition, the coloration of the coat considered to originate from this red flavor was observed in males and females of the test substance-administered group between the 2nd and 4th days. But no abnormality considered to be a toxicity change was observed. No abnormality was found in body weight of animals. Diaphragmatic hernia in the thoracic cavity was found in one female in the 1000 mg/kg group. In the hernia, the caudate portion of the liver protruded nodularly into the thoracic cavity, a part of which was adhered to the chest wall. Since this change was expressed only in one case and not related to the dose, it was judged as a contingent finding. No other abnormality was found. Therefore,LD50 was considered to be >2000 mg/kg bw, when Male and female SD rats were treated with test chemical via oral route.

2.Acute oral toxicity study was done infemale Sprague Dawley rats using test chemical.Distilled water was used as vehicle.Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Step - II). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality after the dosing. No mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - I). Administration of the test item at 2000 mg/kg resulted in diarrhoea (black colour stools) in all animals with onset at 2 hours and no mortality after the dosing. As no mortality were observed at 24 hours after the dosing, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - II). Administration of the test item at 2000 mg/kg resulted in diarrhoea (black colour stools) in all animals with onset at 4 hours and no mortality after the dosing. All animals from 300 mg/kg and 2000 mg/kg dose groups survived through the study period of 14 days. Staining of the stool is attributed to the black colour of the test item. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.Hence,LD50 value was considered to be5000 mg/kg bw,when female Sprague Dawleyrats were treated with test chemical orally via gavage following 14 days of observation period according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

3.In acute oral toxicity study, mice were treated with test chemical orally. 50 % Mortality was observed in treated mice at 20,000 mg/kg bw. Therefore, LD50 was considered to be 20,000 mg/kg bw. When mice were treated with test chemical orally.

Thus, based on the above summarised studies,2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3)and it’s structurallyand functionally similarread across substance, it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation,2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3)cannot be classified for acute oral toxicity.Hence,based on the data available for the structurally and functionally similar read across, test chemical 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3) is not likely to be toxic atleast at the dose of >20000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is Klimisch 1 and data is from secondary study

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Quality of whole database:

Waiver

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is Klimisch 1 and data is from experimental report

Additional information

Acute oral toxicity:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the acute oral toxicity of the test chemical 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3).The studies are as mentioned below:

1.Acute oral toxicity study of test chemical was conducted in 40 Crj: CD (SD) IGS, SPF male and female rat at the concentration of 0, 500, 1000 and 2000 mg/kg bw. The test substance (Purity - >99 %; Obtained from - Dainippon Ink & Chemicals, Inc. and lot number - 000207) was suspended in a medium containing 0.1% Tween 80 added 0.5% CMC-Na aqueous solution as 20 mL/kg. Mortality and general condition were observed over 4 days, 30 minutes, 1, 3 and 6 hours after administration, once a day for 14 days, thereafter. Body weight was measured using an epple dish balance on days 4, 8 and 15 immediately before administration.Animals were observed for clinical signs. No death occurred in both males and females.Clinical signs observed such as, red feces exhibiting the same color tone as the test substance were found between 6 hours and 3 days after administration in males and females of the test substance - administered group, and in all the sexes on the 2nd day. In addition, the coloration of the coat considered to originate from this red flavor was observed in males and females of the test substance-administered group between the 2nd and 4th days. But no abnormality considered to be a toxicity change was observed. No abnormality was found in body weight of animals. Diaphragmatic hernia in the thoracic cavity was found in one female in the 1000 mg/kg group. In the hernia, the caudate portion of the liver protruded nodularly into the thoracic cavity, a part of which was adhered to the chest wall. Since this change was expressed only in one case and not related to the dose, it was judged as a contingent finding. No other abnormality was found. Therefore,LD50 was considered to be >2000 mg/kg bw, when Male and female SD rats were treated with test chemical via oral route.

2.Acute oral toxicity study was done infemale Sprague Dawley rats using test chemical.Distilled water was used as vehicle.Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Step - II). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality after the dosing. No mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - I). Administration of the test item at 2000 mg/kg resulted in diarrhoea (black colour stools) in all animals with onset at 2 hours and no mortality after the dosing. As no mortality were observed at 24 hours after the dosing, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - II). Administration of the test item at 2000 mg/kg resulted in diarrhoea (black colour stools) in all animals with onset at 4 hours and no mortality after the dosing. All animals from 300 mg/kg and 2000 mg/kg dose groups survived through the study period of 14 days. Staining of the stool is attributed to the black colour of the test item. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.Hence,LD50 value was considered to be5000 mg/kg bw,when female Sprague Dawleyrats were treated with test chemical orally via gavage following 14 days of observation period according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

3.In acute oral toxicity study, mice were treated with test chemical orally. 50 % Mortality was observed in treated mice at 20,000 mg/kg bw. Therefore, LD50 was considered to be 20,000 mg/kg bw. When mice were treated with test chemical orally.

Thus, based on the above summarised studies,2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3)and it’s structurallyand functionally similarread across substance, it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation,2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3)cannot be classified for acute oral toxicity.Hence,based on the data available for the structurally and functionally similar read across, test chemical 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3) is not likely to be toxic atleast at the dose of >2000 mg/Kg bw.

Acute inhalation toxicity:

2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated has very low  vapor pressure (2.0265E-17 Pa), so the potential for the generation of inhalable vapours is very low. Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point for repeated inhalation toxicity was considered for waiver.

Acute dermal toxicity:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine theAcute dermal toxicityof the test chemical 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3).The studies are as mentioned below:

1.In acute dermal toxicity study,male and female Sprague Dawley rats were treated with test chemical in the concentration of 2000 mg/kg bw by dermal application.Distilled water was used as vehicle. The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.Hence,The LD50 value was considered to be >2000 mg/kg bw,when male and female Sprague Dawley rats were semiocclusively treated with test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

2.In an acute dermal toxicity study, Wistar male and female rats were treated with test chemical in the concentration of 2000 mg/kg bw by dermal application. No mortality and any clinical sign of toxicity were observed throughout the observation period of 14 days in treated rats. The body weight of each animal recorded on day 0, 7th and 14th showed normal increase and there was no significant increase or decrease in weight was recorded. Therefore, LD50 was considered to be > 2000 mg/kg bw,when Wistar male and female rats were treated with 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs by dermal application.

3.In acute dermal toxicity study,rabbits were treated with test chemical in the concentration of 10000 mg/kg bw by dermal application.No mortality was observed in treated rabbits at dose 10000 mg/kg bw.Therefore, LD50 value was considered to be >10000 mg/kg bw,when rabbits were treated with test chemical  by dermal application.

Thus, based on the above summarised studies,2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3)and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3)cannot be classified for acute dermal toxicity.Hence,based on the data available for the structurally and functionally similar read across chemical, 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3) is not likely to be toxic atleast at the dose of >10000 mg/Kg bw.

Justification for classification or non-classification

Based on the above experimental studies on 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3) and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated(85203-90-3) cannot be classified for acute oral and dermal toxicity. For Acute inhalation toxicity wavier was added so, not possible to classify.