Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate

Endpoint summary

Description of key information

Data available for the structurally similar read across chemicals has been reviewed to determine the acute oral toxicity of the test chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7).The studies are as mentioned below:

1.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).

2.A simple acute oral toxicity experiment was conducted prior to DNA damage in pregnant female CD1 mice. Samples of test chemical, was from certified lots approved for food use in Japan and was purchased from Tokyo Kasei Organic Chemicals (Tokyo). CD-1 (ICR) mice were obtained from Charles River Japan, Inc. (Yokohama) at 7 weeks of age and used for the experiments after 1 week of acclimatization. They were fed with commercial pellets MF (Oriental Yeast Industries, Tokyo) and tap waterad libitumthroughout the acclimatization period and the experiment. The animal room was kept at 22 ± 2°C with a 12-h light-dark cycle; the humidity was 30–50%. For the pregnant mouse study, female mice were mated for a period of one to two days. The morning on which copulation plugs were observed was designated day 0 of gestation. On the morning of day 11, 4 mice were randomly assigned to each treatment group. Chemicals were dissolved in distilled water and administered by gavage at the limit dose of 2000 mg/kg (10 ml/kg). The same volume of distilled water was administered to the control mice at the same time. The limit dose, at which no deaths, morbidity, or distinctive clinical signs were observed, was determined by preliminary acute toxicity tests. The Limit dose at which no mortality, morbidity or toxic signs were observed in mice dosed with test chemical was determined to be greater than 2000 mg/kg bw.

Thus, based on the above summarised studies,Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7) and it’s structurallysimilarread 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,Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7)cannot be classified for acute oral toxicity.Hence, based on the data available for the structurally similar read across, test chemicalTetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7)is not likely to be toxic at the dose range of >2000-5000 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

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.and 2. 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.acute toxic class method 2.comet assay

Limit test:

yes

Specific details on test material used for the study:

- Name of the test chemical: Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate
- Molecular formula: C30H27ClN8O17S5.4Na
- Molecular weight: 1055.29 g/mol
- Substance type: Organic
- Smiles: [Na+].[Na+].[Na+].[Na+].c1c(c(c(c2c(cc(cc12)S(=O)(=O)[O-])NC(CC)=O)O)\N=N\c1c(ccc(c1)Nc1nc(nc(n1)Cl)Nc1cc(ccc1)S(=O)(=O)CCOS(=O)(=O)[O-])S(=O)(=O)[O-])S(=O)(=O)[O-]
- Inchi: 1S/C30H27ClN8O17S5.4Na/c1-2-24(40)34-21-14-19(58(44,45)46)10-15-11-23(60(50,51)52)26(27(41)25(15)21)39-38-20-13-17(6-7-22(20)59(47,48)49)33-30-36-28(31)35-29(37-30)32-16-4-3-5-18(12-16)57(42,43)9-8-56-61(53,54)55;;;;/h3-7,10-14,41H,2,8-9H2,1H3,(H,34,40)(H,44,45,46)(H,47,48,49)(H,50,51,52)(H,53,54,55)(H2,32,33,35,36,37);;;;/q;4*+1/p-4/b39-38+;;;;

Species:

other: 1. rat 2.mouse

Strain:

other: 1.Sprague-Dawley 2.CD-1

Sex:

female

Details on test animals or test system and environmental conditions:

1.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 :
Female
Mean : 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


2.Source: Charles River Japan, Inc. (Yokohama)
Age at study initiation: 7 weeks
Weight at study initiation: no data
Fasting period before study: no data
Housing: no data
Diet (e.g. ad libitum): commercial pellets MF (Oriental Yeast Industries Co., Tokyo, Japan), ad libitum
Water (e.g. ad libitum): Tap Water, ad libitum
Acclimation period: 1 week of acclimatization

ENVIRONMENTAL CONDITIONS
Temperature (°C): The animal room was at 22 ± 2°C
Humidity (%): 30 -50%
Air changes (per hr): No data
Photoperiod (hrs dark / hrs light): 12 h
Light – dark cycle.

Route of administration:

other: 1.oral: gavage 2.oral: gavage

Vehicle:

other: 1.Distilled water 2.Distilled water

Details on oral exposure:

1.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 data

MAXIMUM DOSE VOLUME APPLIED: 10 ml per kg of body weight.

DOSAGE PREPARATION (if unusual): No data

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: No data


2.MAXIMUM DOSE VOLUME APPLIED:
The limit dose of 2000 mg/kg(10 ml/kg)

Doses:

1.Dose Group I : 300 mg/kg
Dose Group I : 300 mg/kg
Dose Group II : 2000 mg/kg
Dose Group II : 2000 mg/kg

2.The limit dose of 2000 mg/kg (10 ml/kg)

No. of animals per sex per dose:

1.Three females were used at each step.

2.4 mice were randomly assigned to treatment group

Control animals:

not specified

Details on study design:

1.- 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.

2.no data

Statistics:

No data

Preliminary study:

No data

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:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality was observed

Mortality:

1.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.
2.no mortality observed

Clinical signs:

other: 1.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 a

Gross pathology:

1.Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.
2.no data

Other findings:

No data

Interpretation of results:

other: not classified

Conclusions:

The test chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate (80019-42-7) is not likely to be toxic atleast in the dose range of >2000-5000 mg/kg bw.

Executive summary:

Data available for the structurally similar read across chemicals has been reviewed to determine the acute oral toxicity of the test chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7).The studies are as mentioned below:

1.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).

2.A simple acute oral toxicity experiment was conducted prior to DNA damage in pregnant female CD1 mice. Samples of test chemical, was from certified lots approved for food use in Japan and was purchased from Tokyo Kasei Organic Chemicals (Tokyo). CD-1 (ICR) mice were obtained from Charles River Japan, Inc. (Yokohama) at 7 weeks of age and used for the experiments after 1 week of acclimatization. They were fed with commercial pellets MF (Oriental Yeast Industries, Tokyo) and tap waterad libitumthroughout the acclimatization period and the experiment. The animal room was kept at 22 ± 2°C with a 12-h light-dark cycle; the humidity was 30–50%. For the pregnant mouse study, female mice were mated for a period of one to two days. The morning on which copulation plugs were observed was designated day 0 of gestation. On the morning of day 11, 4 mice were randomly assigned to each treatment group. Chemicals were dissolved in distilled water and administered by gavage at the limit dose of 2000 mg/kg (10 ml/kg). The same volume of distilled water was administered to the control mice at the same time. The limit dose, at which no deaths, morbidity, or distinctive clinical signs were observed, was determined by preliminary acute toxicity tests. The Limit dose at which no mortality, morbidity or toxic signs were observed in mice dosed with test chemical was determined to be greater than 2000 mg/kg bw.

Thus, based on the above summarised studies,Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7) and it’s structurallysimilarread 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,Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7)cannot be classified for acute oral toxicity.Hence, based on the data available for the structurally similar read across, test chemicalTetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7)is not likely to be toxic at the dose range of >2000-5000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Quality of whole database:

Data is Klimisch 2 and from study report

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Data available for the structurally similar read across chemicals has been reviewed to determine the acute oral toxicity of the test chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7).The studies are as mentioned below:

1.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).

2.A simple acute oral toxicity experiment was conducted prior to DNA damage in pregnant female CD1 mice. Samples of test chemical, was from certified lots approved for food use in Japan and was purchased from Tokyo Kasei Organic Chemicals (Tokyo). CD-1 (ICR) mice were obtained from Charles River Japan, Inc. (Yokohama) at 7 weeks of age and used for the experiments after 1 week of acclimatization. They were fed with commercial pellets MF (Oriental Yeast Industries, Tokyo) and tap waterad libitumthroughout the acclimatization period and the experiment. The animal room was kept at 22 ± 2°C with a 12-h light-dark cycle; the humidity was 30–50%. For the pregnant mouse study, female mice were mated for a period of one to two days. The morning on which copulation plugs were observed was designated day 0 of gestation. On the morning of day 11, 4 mice were randomly assigned to each treatment group. Chemicals were dissolved in distilled water and administered by gavage at the limit dose of 2000 mg/kg (10 ml/kg). The same volume of distilled water was administered to the control mice at the same time. The limit dose, at which no deaths, morbidity, or distinctive clinical signs were observed, was determined by preliminary acute toxicity tests. The Limit dose at which no mortality, morbidity or toxic signs were observed in mice dosed with test chemical was determined to be greater than 2000 mg/kg bw.

Thus, based on the above summarised studies,Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7) and it’s structurallysimilarread 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,Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7)cannot be classified for acute oral toxicity.Hence, based on the data available for the structurally similar read across, test chemicalTetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7)is not likely to be toxic at the dose range of >2000-5000 mg/kg bw.

Justification for classification or non-classification

Based on the above experimental studies on Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate(80019-42-7) and it’s structurally 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,Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate (80019-42-7) cannot be classified for acute oral toxicity.